CN107136219A - The manufacture method of natural cheese - Google Patents

Abstract

The present invention relates to the manufacture method of natural cheese.The present invention provides a kind of manufacture method of curing type natural cheese, it is characterised in that the cheese curd after whey and/or cheese addition lactic acid bacteria have been discharged by relative, so that cell concentration reaches 10⁷Cured after individual/more than g, curing start after soluble nitrogen content (%) reach that the curing time of institute's definite value foreshortens to 20~80% with the occasion added without the lactic acid bacteria compared with, thus cure start after by the soluble nitrogen content [%] at 3 months be more than 3.5%.

Description

How to make natural cheese

The present invention is a divisional application of the invention application with the application number 200880105342.2 (international application number PCT/JP2008/065388), the filing date being August 28, 2008, and the invention title being "Method for Manufacturing Natural Cheese".

technical field

The present invention relates to a natural cheese whose aging time is shortened compared with the existing method, and whose flavor is equal to or better than the existing product. Production method.

Background technique

Examples of the aged cheese include Chaudar, Gouda, Eddam, Emmentale, Parmesan, Camembert, and Feta cheese. In this aged cheese, flavor is mainly formed by the enzyme reaction during aging. Furthermore, this enzymatic reaction involves the following enzymes.

(A-1) Enzymes derived from raw milk (enzymes derived from thermostable bacteria, etc. which remain after sterilization of raw milk)

(A-2) Enzymes (peptidase, aminopeptidase, etc.) derived from lactic acid bacteria

(A-3) Enzymes derived from rennet

(A-4) Enzymes derived from microorganisms other than lactic acid bacteria such as molds (cheese using molds, etc.)

Here, in the case of cheese using molds, etc., enzymes from microorganisms such as the above-mentioned (A-4) molds have the greatest influence on the formation of flavors, but in the case of other cheeses, enzymes from the above-mentioned (A-2) lactic acid bacteria are usually used. Enzymes have a great influence on the formation of flavor during ripening.

The flavor of cheese is produced by decomposing protein, carbohydrates, and fat as its components through an enzymatic reaction. At this time, when it is a protein, it will generate amino acids, amines, sulfides, etc.; when it is a carbohydrate, it will generate lactic acid, ethanol, formaldehyde, etc.; flavor substances, aroma substances, taste). Moreover, since the production amount and type of enzymes differ depending on the species and strain of lactic acid bacteria (fermentation material), the production amount and type of flavor components also differ, and the flavor of cheese varies with different lactic acid bacteria. Therefore, a producer of lactic acid bacteria (a starter supplier) prepares a variety of starters when manufacturing the same type of cheese, and a cheese maker (cheese maker) selects and uses a starter by their own judgment.

On the other hand, the flavor of cheese is of course also affected by the manufacturing conditions (chemical conditions, physical conditions, etc.) of cheese. This is due to changes in the growth of lactic acid bacteria, differences in the production amount and types of enzymes, or changes in enzyme reactions, and changes in the concentration and types of trace components in cheese. Here, the manufacturing conditions of cheese include the component composition of raw material milk (milk for cheese), the temperature and pH at the time of curd preparation, aging temperature, aging time (aging speed), etc. Of course, most cheese makers (manufacturers) choose the production conditions of cheese based on historical traditions and experience rather than selecting the production conditions of cheese according to their own judgment. Also, it can be considered that this selection of manufacturing conditions is optimal.

Therefore, the prototype (original) of most modern natural cheeses is made in Western Europe, and the flavor and eating methods of the cheese are closely connected with Western European food culture. That is, for most cheeses, the flavor and the like that match the food culture of Western Europe are selected, and the cheese production conditions are further selected. Conversely, since the food culture in Japan is different from that in Western Europe, all cheeses may not be accepted by ordinary consumers in Japan when they use their original manufacturing conditions and flavors. For example, with regard to the flavor of aged cheese, Japan differs from Western Europe in that the flavor components (taste) derived from fatty acids and the like are weak and stable, and the flavor, especially aroma, is strong and tends to be preferred. For cheese makers, the flavor of aged cheese does not completely match the preferences of Japanese consumers. From the perspective of improving the marketability of products, this is a matter of great concern. Considering its flavor and preferences, the emphasis on fragrance ( strengthen). On the other hand, aging of cheese requires a long period of time (many days), and installation costs and operating costs for its storage facilities (aging warehouses) are required. At this time, such as promoting the ripening of cheese and increasing the ripening speed, it is also possible to reduce the operating cost (storage fee) of the ripening warehouse. Therefore, various methods were tried.

The aging of natural cheese is mainly carried out by the enzymes produced by the starter lactic acid bacteria. Associated with aroma and structure are proteolytic enzymes (proteases). Proteases are classified into endo-proteases and exo-proteases based on their mechanism of action. Endoproteases cleave peptide bonds located inside protein molecules. The exoprotease acts from the N-terminal side and the C-terminal side of the low-molecular-weight peptide to cleave amino acids and dipeptides. When the lactic acid bacteria remain, the enzymes are released from the outside of the bacteria (the enzymes outside the bacteria, mainly endoproteases), and when they die during aging, the enzymes in the bacteria (mainly exoproteases) are released through the action of bacteriolysis. The production of aroma (amino acid) is mainly exoprotease. Generally, lactic acid bacteria used in cheese produce lactic acid, increase the effect of rennet, promote the formation of curds, release enzymes during aging, and contribute to flavor formation and texture softening.

Non-Patent Document 1 describes a method of adjusting the flavor of cheese or controlling the aging rate. That is, there are (B-1) control (improvement) of aging rate, (B-2) use of proteases other than lactic acid bacteria, (B-3) use of ajankto starter, (B-4) genetically recombinant starter The use of (B-5) the use of high-pressure treated starter, etc. In the above (B-1), the speed of the enzyme reaction is controlled. However, since the curing temperature is generally at the upper limit (about 15° C.), the effect of adjusting the flavor of cheese or increasing the curing speed is also limited. In the above (B-2), as enzyme preparations derived from proteases other than lactic acid bacteria, enzyme extracts derived from molds and the like can actually be purchased. However, when enzymes derived from molds are used, the cheese tends to have a bitter taste, and the types and flavors of cheese are limited. In the above (B-3), in addition to the main starter, an ajunct starter may be used in combination. A method such as not being used in the main starter was selected due to reasons such as the reduction of the acid production ability of the ajunct starter. In the above-mentioned (B-4), by genetic recombination operation, the starter which improves the enzyme production ability of a lactic acid bacterium, etc. can be used. However, the kind of starter is limited, and the kind and flavor of cheese are limited. The above-mentioned (B-1) and (B-3) can be used in actual production, but the above-mentioned (B-2) and (B-4) are hardly applicable to actual production and are not practical.

In addition, in order to promote the ripening of cheese, Non-Patent Document 2 discloses that in a tank (cheese vat) for cheese production, a certain amount of lactic acid bacteria starter is added to raw milk (milk for cheese) and kept at a temperature of 30° C. and a pH of 6.6 Fermentation (cultivation) is carried out for about 10 hours, and the method of ripening after the lactic acid bacteria of cheese milk reaches a high concentration. However, since cheese milk is kept in a cheese vat for a long time while controlling the temperature and pH of the cheese milk, the production efficiency is very poor in mass production and continuous production, which is not practical. In addition, keeping at about 30°C for a long time may cause problems such as bacterial proliferation (bacterial contamination) and quality deterioration (increase in acidity) of acid produced by lactic acid bacteria (fermentation).

Patent Document 1 discloses a method of subjecting a culture of lactic acid bacteria (a ferment, etc.) to pressure treatment to lose the acid-generating ability of the lactic acid bacteria, and using the culture as an enzyme preparation in combination with the ferment. However, in order to control the activity of lactic acid bacteria, so-called pressure treatment has been added. In addition, in order to add an enzyme preparation to raw material milk (cheese milk), there is a danger that many of the active ingredient enzymes will be discharged together with the whey. That is, only a part of the actually added enzyme is kept in the cheese curd and used, and the loss (loss) of the enzyme of the active ingredient is large.

Patent Document 2 discloses a method of using an enzyme derived from a specific mold and an enzyme derived from a specific lactic acid bacterium in combination. However, in order to use specific molds and lactic acid bacteria, the technical versatility is low. However, in order to treat the culture of lactic acid bacteria with an enzyme (lysozyme), a so-called means of reacting with the enzyme is added. Moreover, since cheese tends to produce a bitter taste in order to use the enzyme derived from a mold, there exists a possibility that the kind and flavor of cheese may be limited.

In recent years, in order to accelerate aging of cheese, a method of extracting proteolytic enzymes of microorganisms such as molds and introducing them into cheese production has been studied (Patent Document 3, Patent Document 4). In addition, proteolytic enzymes derived from microorganisms for food use are also sold, not limited to the acceleration of aging of cheese.

However, there are few commercially available cases in which proteolytic enzymes derived from microorganisms enhance the flavor of natural cheese or promote ripening. Regarding the utilization of cheese, there are few cases where it is used in the production of EMC (enzyme-improved cheese; cheese flavor enhancer). The reason why it is not practical in the flavor enhancement or accelerated maturation of natural cheese is due to the generation of bitterness. In the case of EMC, since the decomposition of protein is promoted compared with cheese, the bitter peptide is further decomposed and the bitterness is reduced.

The bitter taste of cheese is produced by the formation of bitter peptides during the decomposition of protein. Generally, the exoprotease from lactic acid bacteria decomposes into amino acids from the end of the protein molecular chain connected by multiple amino acids. Most of the proteolytic enzymes on the market come from microorganisms. The exoprotease has high activity in decomposing protein molecular chains, so it is easy to produce bitterness.

Patent Document 1: Japanese Unexamined Patent Publication No. 3-160944

Patent Document 2: JP-A-7-236484

Patent Document 3: JP-A-3-160944

Patent Document 4: Japanese Unexamined Patent Publication No. 7-236484

Non-Patent Document 1: CHEESE Chemistry, Physics and Microbiology, Third Edition (Fox) P.289, 2004

Non-Patent Document 2: Netherlands Milk & Dairy Journal1, Vo1.15, 1961

Contents of the invention

Problems to be solved by the present invention

The present invention is proposed in view of the problems of the above-mentioned prior art, and the purpose of the present invention is to provide a natural cheese production method that can effectively control or adjust aging time and flavor (especially aroma) through simple operations.

Another object of the present invention is to provide an aging-type natural cheese manufacturing method that is particularly suitable for mass production or continuous production, enhances aroma while promoting aging, and can suppress bitterness.

Another object of the present invention is to provide: improved flavor of matured natural cheese, almost no bitterness, strong cheese flavor such as aroma, and a cheese that achieves a predetermined matured flavor and texture in a short time and its manufacturing method.

The means used to solve this problem

In view of the above-mentioned problems, the present inventors have conducted intensive studies and found that the aging time and flavor can be effectively controlled and adjusted through simple operations by devising methods for the enzymes involved in the enzymatic reaction during aging and the conditions for adding them, and completed the present invention. .

That is, it was found that the addition conditions of lactic acid bacteria and their crushed bacteria-derived processed products that produce enzymes involved in the enzyme reaction during aging, or the addition conditions of proteolytic enzymes derived from microorganisms, were found in the existing natural cheese production process. Through simple operation, the aging time and flavor can be effectively controlled and adjusted, and the invention has been completed.

At this time, it was also found that in the natural cheese production process, lactic acid bacteria or their crushed bacteria were added to cheese curds or cheese to increase the concentration of bacteria and then ripened, and the enzymes derived from lactic acid bacteria were intentionally increased and utilized. A good flavor (in particular, aroma) is formed while promoting ripening, and aged cheese is obtained.

In addition, the present inventors have hypothesized and carefully studied the reason why the bitter taste tends to appear from the proteolytic pattern of the enzyme. As a result, it was discovered that by aging cheese with added enzymes, grasping the protein decomposition status during the aging process, selecting the type and amount of enzymes, and setting aging conditions, a method for obtaining a good aging flavor without bitterness was obtained.

Specifically, after adding (addition, inoculation, etc.) any of lactic acid bacteria, lactic acid bacteria crushed cells, and proteolytic enzymes derived from microorganisms to cheese curds before aging or cheese during aging, aging is started or continued . Alternatively, add (addition, inoculation, etc.) two types of lactic acid bacteria and lactic acid bacteria crushed cells, two types of lactic acid bacteria and microorganism-derived proteolytic enzymes, two types of lactic acid bacteria crushed cells and microorganism-derived proteolytic enzymes, and lactic acid bacteria Aging starts or continues after the crushed lactic acid bacteria and any of proteolytic enzymes derived from microorganisms. Through these operations, the aroma of cheese appears rapidly, and the aroma of cheese is enhanced, and as a result, aging of cheese is accelerated and bitterness is suppressed.

At this time, it has been found that it is preferable to add lactic acid bacteria, protein crushed bacteria, proteolytic enzymes derived from microorganisms, and the like to the cheese curd and/or cheese from which the whey has been drained. When doing so, lactic acid bacteria, the cell crushed material of lactic acid bacteria, proteolytic enzyme derived from microorganisms, etc. can be directly added to cheese curd or cheese. Here, enzymes derived from lactic acid bacteria or proteolytic enzymes derived from microorganisms which are active ingredients thereof are not discharged together with whey. That is, all or most of the actually added enzymes derived from lactic acid bacteria or proteolytic enzymes derived from microorganisms are retained in the cheese curd and used, and the enzymes that are active ingredients thereof are not lost or slightly lost (lost).

Usually, when adding lactic acid bacteria (live bacteria) to the fermented product, when adding lactic acid bacteria to raw milk (cheese milk), for example, because the pH of the raw milk is too low, the cheese curd is too hard, the moisture is too low, and the sour taste is too strong. Affects the state of cheese curd production. However, when lactic acid bacteria or proteolytic enzymes derived from microorganisms were directly added to cheese curd, there was no influence on the production state of curd.

However, lactic acid bacteria (living bacteria) are originally lysed while slowly dying in cheese curd during aging of cheese. However, as a result, the enzymes in the cells are released outside the cells, and the cheese components are decomposed by the action of these enzymes. Therefore, in order for the enzymes in the cells to act on the cheese components, it takes time necessary to achieve lysis. Conversely, when lactic acid bacteria are crushed to destroy the cell wall of lactic acid bacteria, first, the enzymes in the bacteria are released from the outside of the bacteria, and from the moment of adding to cheese curd or cheese, they act on the cheese components. Compared with adding lactic acid bacteria with live bacteria , the time is short, which can effectively promote the ripening. Based on these findings, it has been found that the aging time is shorter than the existing method, and the flavor is equal or better than the existing product. On the other hand, if the aging time is the same as the existing method, the flavor is good compared to the existing product ( Especially when the flavor is emphasized), the production method of natural cheese.

In the present invention, since lactic acid bacteria, lactic acid bacteria crushed cells, proteolytic enzymes derived from microorganisms, etc. are directly added to cheese curd or cheese, the flavor of cheese can be adjusted during aging. Here, when cheese curd is formed, instead of the lactic acid bacterium starter, or together with the lactic acid bacterium starter, you may use a pH adjuster. In this case, as a pH adjuster, gluconolactone (GDL) is mentioned. Gluconolactone gradually lowers the pH of raw material milk (cheese milk) at a predetermined time, and forms cheese curd with predetermined physical properties (hardness). When the pH regulator was used instead of the lactic acid bacteria starter, the flavor and physical properties of cheese curd had no effect on the lactic acid bacteria starter. Here, in aged natural cheese, flavor and physical properties can be adjusted arbitrarily by adding lactic acid bacteria, crushed lactic acid bacteria, proteolytic enzymes derived from microorganisms, etc. to cheese curd.

That is, the present invention is as follows.

The method for producing natural cheese according to the invention of claim 1, wherein lactic acid bacteria are added to the cheese curd and/or cheese from which the whey has been drained so that the bacterial cell concentration becomes 10 ⁷ cells/g or more, and then aged.

The invention of claim 2 relates to the method for producing natural cheese according to claim 1, wherein the lactic acid bacteria are live bacteria.

The method for producing natural cheese according to the invention of claim 3, wherein the processed product of lactic acid bacteria is added to the cheese curd and/or cheese after the whey has been discharged so that the bacterial cell concentration corresponds to 10 ⁵ cells/g After the above, ripening is carried out.

The invention of claim 4 relates to the method for producing natural cheese according to claim 3, wherein the bacterial cell crushing treatment is homogenization treatment at an operating pressure of 100 MPa or more.

The invention of claim 5 relates to the method for producing natural cheese according to any one of claims 1 to 4, wherein lactic acid bacteria concentrated by any one of membrane separation, centrifugal separation, and vacuum evaporation are used.

The invention of claim 6 relates to the method for producing natural cheese according to any one of claims 1 to 4, characterized in that the cheese dried by any one of freeze-drying, reduced-pressure spray-drying, and spray-drying is used. Lactic acid bacteria.

The invention of claim 7 relates to the method for producing natural cheese according to any one of claims 1 to 6, characterized in that neutralized cultured lactic acid bacteria are used.

The invention of claim 8 relates to a method for producing natural cheese, characterized in that the cheese curd and/or cheese from which the whey has been drained is added with proteolytic enzymes derived from microorganisms, and then aged.

The invention of claim 9 relates to the method for producing natural cheese according to claim 8, wherein the contribution of Phospho Tangsten Acid Soluble Nitrogen (PTASN) to the total When total nitrogen (TN) reaches 6.0-8.5% (PTASN/TN×100), the ratio of water soluble nitrogen (WSN) to PTASN (WSN/PTASN) is below 4.5.

The invention of claim 10 relates to the method for producing natural cheese according to the invention of claim 8 or 9, wherein the proteolytic enzyme derived from microorganisms has a protease activity of 2000 unit/g or more and a peptidase activity of 160 unit/g or more .

The invention of claim 11 relates to the method for producing natural cheese according to any one of claims 1 to 10, wherein the cheese curd and/or cheese from which the whey has been drained are not sterilized.

The invention of claim 12 relates to the method for producing natural cheese according to any one of claims 1 to 11, wherein the cheese curd is formed by adding a pH regulator to raw material milk.

The invention of claim 13 relates to the method for producing natural cheese according to claim 12, wherein the pH adjuster is gluconolactone.

The invention of claim 14 is natural cheese produced by the production method described in any one of claims 1 to 13.

The effect of the invention

According to the present invention, there is provided a method for producing natural cheese that effectively controls and adjusts aging time and flavor (especially aroma) with simple operations.

Also, according to the present invention, there is provided a method for producing aged natural cheese which is especially suitable for mass production and continuous production, and which enhances flavor while promoting ripening and suppresses bitterness.

In addition, according to the present invention, there are provided: improved flavor of aged natural cheese, cheese flavor such as almost no bitterness, strong aroma, etc., achieving predetermined aged flavor and texture in a short time, and a method for producing the same.

Description of drawings

Fig. 1 is a graph showing the soluble nitrogen content of conventional Gouda cheese after aging has started.

Fig. 2 is a graph showing the soluble nitrogen content of the cheese of the present invention after the initiation of aging.

Fig. 3 is a graph showing the enzyme activity of the cheese of the present invention after ripening.

Fig. 4 is a graph showing the soluble nitrogen content of Gouda cheese according to the present invention after aging starts.

Fig. 5 is a graph showing the soluble nitrogen content of Gouda cheese according to the present invention after aging starts.

Fig. 6 is a graph showing the soluble nitrogen content of soda cheese according to the present invention (addition of enzyme) after ripening.

detailed description

The inventors of the present invention studied flavor components (taste) produced by enzymatically decomposing cheese curds or cheese components (proteins, carbohydrates, fats, etc.) during aging in aged cheeses.

However, in the existing natural cheese production process, aging-type cheeses are trial-produced under various addition conditions such as lactic acid bacteria or their crushed bacteria that produce enzymes during aging, proteolytic enzymes derived from microorganisms, and these cheeses are evaluated. , Analysis of taste or flavor, physical properties, etc. In this case, a sensory test was used for the evaluation of the food texture and flavor, and the soluble nitrogen content was used as an indicator of aging progress for the evaluation of the physical properties. By simultaneously evaluating and analyzing these textures, flavors, and physical properties, an effective method for producing aged cheese with good textures or flavors has been discovered.

That is, through experimental research, it was found that in the existing natural cheese manufacturing process, the addition conditions of lactic acid bacteria or their crushed bacteria, proteolytic enzymes from microorganisms, etc. can be effectively controlled through simple operations. And adjust the aging time and flavor. At this time, in the process of producing natural cheese, it was also found that adding high-concentration lactic acid bacteria to cheese curds or cheese and then aging them, intentionally increasing and utilizing enzymes derived from lactic acid bacteria, and improving flavor (especially enhancing aroma) while promoting aging ), a method for obtaining aged cheese. In addition, it has also been found that by aging cheese to which proteolytic enzymes from microorganisms have been added, by grasping the state of proteolysis during the aging process, by selecting the type and amount of enzyme added, and setting aging conditions, no bitterness can be obtained, and a good aging flavor can be obtained. Method of fortified aged cheese.

The method for producing natural cheese of the present invention is characterized in that lactic acid bacteria are added to the cheese curd and/or cheese from which the whey has been drained so that the bacterial cell concentration becomes 10 ⁷ cells/g or more, preferably 10 ⁸ cells/g The above is ripened.

At this time, if the addition of lactic acid bacteria to cheese curd and/or cheese is less than 10 ⁷ /g, the effect of the present invention of greatly shortening the aging time or improving flavor (especially when flavor is emphasized) cannot be sufficiently obtained.

In addition, when lactic acid bacteria are added to cheese curd and/or cheese, the number of lactic acid bacteria does not reach a specific upper limit, and it is best to set it while investigating the degree of effect of the present invention intended to be actually used, the flavor of cheese, and the efficiency of the manufacturing process. .

By adding lactic acid bacteria to cheese curd and/or cheese at a high concentration, it is easy to shorten the aging time and improve (intensify) flavor. However, when lactic acid bacteria are added in excess, undesired flavors other than aroma may also be enhanced.

Therefore, as the upper limit of the number of lactic acid bacteria, for example, 10 ⁹ cells/g, 10 ¹⁰ cells/g, 10 ¹¹ cells/g, etc. are considered. On the other hand, as the aging of cheese progresses, the number of viable lactic acid bacteria decreases while lysing the bacteria, the enzymes in the bacteria are released outside the bacteria, and the cheese components are converted into flavor substances (smell, etc.).

During aging of cheese, it is generally believed that since lactic acid bacteria are undergoing generation renewal, they cannot actively and continuously carry out metabolic activities such as acid production, so as long as no excessive addition of lactic acid bacteria is performed, no special defects are found except for the effect of promoting ripening.

In the present invention, the cheese curd from which the whey has been drained is the cheese curd before aging, and the cheese corresponds to the cheese at the initial stage of aging. The so-called cheese at the beginning of aging means, for example, cheese that has been molded and packaged once and then transferred to the aging process, or cheese that is in the aging process but has not reached a predetermined degree of aging (aging time) (usually less than 1/3).

In addition, when the manufacturing method of the natural cheese of this invention is used, lactic acid bacteria may be live bacteria or dead bacteria. That is, it is not necessary to intentionally reduce the acid-generating ability (activity) of lactic acid bacteria originally possessed, and the lactic acid bacteria do not need to be specially treated by any method, and live bacteria can also be used as they are. At this time, when live lactic acid bacteria are directly added to cheese curd or cheese, the number of live bacteria decreases with time and lysis occurs, and the enzymes in the bacteria are released from the outside of the bacteria and act on the cheese together with the enzymes outside the bacteria. Curd or cheese, to aid ripening or enhance flavor.

In addition, in order to increase the number of lactic acid bacteria in cheese curd or cheese, live bacteria (fermentation products, etc.) The acidity of milk rises too much, so it is necessary to think of a special method for pH control technology.

On the other hand, even if the dead bacteria of lactic acid bacteria are directly added to cheese curd or cheese, if the enzymes in the bacteria are not inactivated, they will act on cheese curd or cheese together with any enzymes released outside the bacteria to promote ripening or enhance the fragrance.

In addition, lactic acid bacteria are subjected to physical (mechanical) or chemical thalline crushing treatment, as the cell wall or cell membrane is broken or destroyed, the enzyme in the thalline is forced to release outside the thalline, or the enzyme in the thalline is to the cheese curd or The time it takes for cheese to start working is shortened.

Therefore, in the method for producing natural cheese of the present invention, to the cheese curd and/or cheese after the whey is discharged, the bacterial cell crushing treatment of lactic acid bacteria is added so that the bacterial cell concentration corresponds to 10 ⁵ cells/g or more, and preferably corresponds to 10 cells/g. Aging may be performed after ⁶ pieces/g or more.

At this time, the addition of lactic acid bacteria cell crushing treatment, so that the cell concentration corresponds to 10 ⁵ cells/g or more, means, for example, after the cell crushing treatment of lactic acid bacteria cultures with a cell concentration of 10 ⁷ cells/g or more 1 weight % or more is added to the cheese curd with the culture solution processed by the crushing of the bacteria, so that the cheese curd contains the crushed lactic acid bacteria cells whose cell concentration is equivalent to 10 ⁵ cells/g or more.

That is, when the culture of lactic acid bacteria with a cell concentration of ¹⁰⁷ /g or more is subjected to cell crushing treatment, since the lactic acid bacteria form cell sheets (fragments) and the like and do not maintain the cell shape, the number of lactic acid bacteria cannot be accurately measured, but the number of lactic acid bacteria is substantially Fragments of lactic acid bacteria at a body concentration corresponding to 10 ⁷ cells/g or more are contained in the culture solution. A culture solution containing lactic acid bacteria fragments, etc., at a cell concentration corresponding to 10 ⁷ cells/g or more, when 1% by weight or more is added to cheese curd or cheese, the cell concentration in cheese curd or cheese is equivalent to 10 ⁵ The bacterial cell crushing of lactic acid bacteria more than one/g. However, the cell crushed product of the lactic acid bacteria acts on cheese curd or cheese to promote ripening or enhance flavor.

In addition, when the manufacturing method of the natural cheese of this invention is used, dry crushing, wet crushing, etc. can be illustrated as a cell crushing process. Specifically, a ball mill, a glass bead mill, a homogenizer (homogenizer), an ultrasonic device, and the like can be used. More specifically, a hollow particle electric shock generator (hollow particle diameter: 0.1-0.5mm, preferably 0.2-0.4mm, more preferably 0.3mm), a high-pressure homogenizer (operating pressure: 100-200MPa, preferably 130-150MPa, More preferably 140MPa) and the like.

At this time, as the bacterial cell crushing treatment, when its efficiency is investigated, it is particularly preferable to use a high-pressure homogenizer and perform homogenization treatment at an operating pressure of 100 MPa or more because continuous treatment is easy.

In addition, in the present invention, since enzymes derived from lactic acid bacteria can be used, a part of them may remain as live bacteria after the lactic acid bacteria cell crushing treatment.

In the conventional method for producing natural cheese, in order to adjust the aging time, the following various methods can be adopted. Add lactic acid bacteria starter to cheese milk and then ferment it to increase the number of lactic acid bacteria. Utilizes enzymes from molds. Heat treatment, pressure treatment, enzyme (lysozyme) treatment, etc. are performed on the culture of lactic acid bacteria (lactic acid bacteria starter, etc.) to eliminate the acid production ability of lactic acid bacteria, and the culture is used as an enzyme preparation, and the starter is used together.

However, in these manufacturing methods, for example, there are the following problems. Even if lactic acid bacteria (live bacteria) are added to cheese milk, normal cheese curd cannot be produced due to an excessive increase in acidity. Even if the lactic acid bacteria culture-treated enzyme preparation is added to cheese milk, most of it is lost (lost) with whey effluent. Using enzymes from molds, it develops a bitter taste during ripening.

Therefore, in the existing natural cheese manufacturing method, the method of adjusting the aging time has many restrictions (low versatility) and troubles, etc., and there are applicability defects in mass production and continuous production.

When the method for producing natural cheese according to the present invention is adopted, any one of lactic acid bacteria, lactic acid bacteria crushed cells, proteolytic enzymes derived from microorganisms, or these Any composition for solving the problems or problems of the existing production methods, which has the following characteristics.

(C-1) It does not affect the process or operation at the time of curd preparation, and does not need to change the existing process and operation.

(C-2) There is a large degree of freedom in the selection of lactic acid bacteria (fermentation products, etc.) at the time of curd preparation.

(C-3) Unlike when the enzyme preparation is added to the raw material milk (cheese milk), since the enzyme does not flow out with the whey and is not lost, all of its effects can be effectively utilized.

(C-4) Not only dead lactic acid bacteria but also live bacteria can be used, and lactic acid bacteria do not need to be processed into enzyme preparations or the like.

(C-5) Not only live lactic acid bacteria, but also dead bacteria or crushed bacteria can be used. Before adding to cheese curd or curd, the number of viable lactic acid bacteria can also be reduced. Therefore, cultures of lactic acid bacteria, etc., It can be stored in a refrigerated state or in a frozen state (the preservation method of lactic acid bacteria is less restricted and has a high degree of freedom).

(C-6) Not only the cheese curd before aging but also the cheese during aging is applied even in the middle of aging.

In the above (C-6), for example, the cheese is first matured by a conventional method, and in the middle of the aging, lactic acid bacteria, the crushed cells of lactic acid bacteria, and proteolytic enzymes derived from microorganisms are directly added (added) to the cheese. Any one of these, or any combination of these is also acceptable. Specifically, it is conceivable to pulverize the aging cheese with a meat grinder or the like, and to add and mix lactic acid bacteria, lactic acid bacteria crushed cells, proteolytic enzymes derived from microorganisms, or any combination thereof. After the cheese is processed, the cheese is improved, vacuum-packed, and then matured.

In the production method of the present invention, any one of lactic acid bacteria, a crushed product of lactic acid bacteria, proteolytic enzymes derived from microorganisms, or any combination of these is directly added (added) to cheese curd or cheese from which whey has been drained Examples of the lactic acid bacteria in the product include the genus Lactococcus, the genus Lactobaci11us, the genus Streptococcus, the genus Leuconostoc, the genus Propionibacterium, and the genus Bifidobacterium. Specific examples include Lactococcus lactis subsp.lactis, L.lactis subsp.lactis biovar diacetilactis, L.lactis subsp.cremoris, Lactobacillus helveticus, L.helveticus subsp.jugurti, L.delbrueckii subsp.bulgaricus, L.delbrueckii subsp.lactis , L.acidophilus, L.crispatus, L.amylovorus., L.gallinarum, L.gasseri, L.johnsonii, L.casei., L.caseisubsp.rhamnosus, Streptococcus salivarius subsp.thermophilus, Leuconostoccremoris, Leu.lactis, Leu .mesenteroides subsp.mesenteroides, Leu.mesenteroides subsp.dextranicum, Leu.parames enteroides, Propionibacterium shermani, Bifidobacterium bifidum, B.longum, B.breve.B.infantis, B.adolescentis, etc. In this case, these lactic acid bacteria may be used alone or in combination of two or more.

In the method for producing natural cheese of the present invention, the lactic acid bacteria can be used after being concentrated by any one or more methods of membrane separation, centrifugal separation, and vacuum evaporation. In addition, lactic acid bacteria dried by any one of the freeze-drying method, the reduced-pressure spray-drying method, and the spray-drying method can also be used. In addition, neutralized cultured lactic acid bacteria can also be used.

Increase the number of lactic acid bacteria per unit capacity by membrane separation method, centrifugal separation method, vacuum evaporation method, freeze drying method, decompression spray drying method, spray drying method, neutralization culture method, etc. to high concentrations.

At this time, when the culture of lactic acid bacteria is produced by membrane separation method, centrifugal separation method, vacuum evaporation method, neutralization culture method, etc., the number of lactic acid bacteria reaches about 10 ¹⁰ to 10 ¹² /g. When the spray drying method, spray drying method, etc. are used for production, the lactic acid bacteria are about 10 ¹¹ to 10 ¹³ cells/g.

When the method for producing natural cheese of the present invention is used, the aging time is easily shortened and the flavor is easily enhanced by adding lactic acid bacteria to cheese curd or cheese at a high concentration. However, when the culture of lactic acid bacteria is added in excess, the composition and The influence of flavor is increased, and there is a possibility that undesirable physical properties or flavor may be formed. Therefore, it is preferable to add a culture of lactic acid bacteria in small amounts, and to add lactic acid bacteria to cheese curd or cheese at a high concentration. That is, it is preferable to add lactic acid bacteria to cheese curd or cheese after adjusting the concentration of lactic acid bacteria to a high concentration using the above-mentioned concentration method, drying method, neutralization culture method, etc., by a culture of lactic acid bacteria. On the other hand, when the addition of the culture of lactic acid bacteria is too small, lactic acid bacteria may not mix uniformly in cheese curd or cheese, and may deviate from a dispersed state. Therefore, it is appropriate that the addition amount of the culture of lactic acid bacteria reaches 0.1～5 weight %, preferably 0.5～4 weight %, more preferably 1～3 weight % to cheese curd or cheese setting. It is preferable to add milk or cheese so that lactic acid bacteria reach 10 ⁷ /g or more.

According to the method for producing natural cheese of the present invention, it is important that the lactic acid bacteria do not remain and die when the lactic acid bacteria are concentrated or dried, and the enzymes of the lactic acid bacteria are not lost or inactivated. Specifically, lactic acid bacteria must be treated at a low or medium temperature such as 50°C or lower or 40°C or lower. Therefore, in the concentration method of lactic acid bacteria, membrane separation method, centrifugation method are preferred than vacuum evaporation method, in the drying method of lactic acid bacteria, freeze drying method, vacuum spray drying method are preferred than spray drying method.

However, the enzymes possessed by lactic acid bacteria include extracellular enzymes produced (released) outside the bacterial cells and intracellular enzymes produced (retained) in the bacterial cells. However, in the extracellular enzymes, the proteases that decompose proteins into peptides with large molecular weights are the main body, while in the in vivo enzymes, the peptides that enter the bacterial cells are decomposed into peptides or amino acids with small molecular weights or peptidases. Aminopeptidase as the main body. In this case, the chymosin-derived enzyme of rennet acts as the main protease, and the extracellular enzyme derived from lactic acid bacteria is not particularly necessary. On the other hand, peptides with a large molecular weight are factors that contribute to bitter taste, while peptides or amino acids with a small molecular weight are factors that contribute to flavor. In this case, since the peptidase or aminopeptidase is mainly derived from the enzymes in the cells of lactic acid bacteria, the enzymes in the cells are particularly required. Here, the vacuum evaporation method among the concentration methods of lactic acid bacteria can achieve a high concentration of lactic acid bacteria together with the components of the culture solution. However, when the membrane separation method and centrifugal separation method are used, only the cells of the lactic acid bacteria other than the components of the culture solution can be highly concentrated. . That is, when the membrane separation method or centrifugal separation method is used, the aroma of the cheese increases, the ratio of enzymes in the bacteria increases, and the concentration of lactic acid bacteria becomes high, which is more effective than the vacuum evaporation method. In addition, the method for producing natural cheese according to the present invention is more effective when protease (proteolytic enzyme) for food is used in combination appropriately.

In the manufacturing method of the natural cheese of this invention, it illustrates as follows as a neutralization cultivation method. That is, it is a method in which lactic acid bacteria are proliferated at 25 to 40° C. near an appropriate temperature while controlling the pH to about 5 to 6 with the lactic acid bacteria culture solution, and kept for 10 to 36 hours for culturing. In the lactic acid bacteria culture solution, as long as it is a well-known liquid medium for lactic acid bacteria to proliferate (grow), such as skim milk, reduced skim milk, whey, reduced whey, etc., as a carbon source such as glucose, lactose, sucrose, etc., as Nitrogen sources such as yeast extract, meat extract, peptone, etc., as salts such as monopotassium phosphate, dipotassium phosphate, sodium acetate, etc. can be added for preparation. In the pH control at the time of lactic acid bacteria culture, the aqueous solution of sodium hydroxide, ammonia, sodium carbonate, etc. can also be used as an alkaline agent. Add alkaline agent to the lactic acid bacteria culture solution to neutralize the lactic acid that hinders or inhibits the growth of lactic acid bacteria and promote the proliferation of lactic acid bacteria. In the case of neutralization culture, the number of lactic acid bacteria per unit volume was increased to about 10 times compared with the case where neutralization culture was not performed.

The manufacturing method of the natural cheese of this invention is characterized by adding the proteolytic enzyme derived from microorganisms to the cheese curd and/or cheese from which the whey was drained, and aging them.

In this case, it is characterized in that after adding the above-mentioned proteolytic enzyme derived from microorganisms, the phosphotungsten acid soluble nitrogen (Phospho Tangsten Acid Soluble Nitrogen (PTASN)) to the total nitrogen (Total Nitrogen (TN)) of the aged cheese reaches 6.0～ At 8.5% (PTASN/TN×100), the ratio of Water Soluble Nitrogen (WSN) to PTASN (WSN/PTASN) is below 4.5.

In addition, it is characterized in that the proteolytic enzyme derived from the above microorganism has a protease activity of 2000 unit/g or more and a peptidase activity of 160 unit/g or more.

Among the above, phosphotungstic acid soluble nitrogen (PTASN) can be measured by the Keidel nitrogen method, and an example of the operation sequence such as pretreatment is described below. Take 25 g of curd cheese and/or cheese (sample), and dissolve it in 150 ml of warm water. Several drops of formalin (40%) were added thereto, and the mixture was kept at 50° C. for 2 hours while shaking. Then, the residue after removing the fat layer was centrifuged at 3000 rpm for 5 minutes. Filter the supernatant with cotton cloth with a fine mesh, and put the filtrate (through liquid) into a measuring bottle (250ml). After washing the precipitate remaining on the quick-immersion tube and cotton cloth with hot water, the centrifugation and filtration operations were repeated twice, and the liquid obtained in this way was mixed with the original filtrate. Water was added to the mixture to bring the liquid to 250ml. Take 50ml of this liquid, add 10ml of water, 30ml of sulfuric acid (25w/v%) and 10ml of PTA aqueous solution (19w/v%) to it, and keep it at room temperature for 24 hours. It was filtered with filter paper (TOYO No. 5B), and 20 ml of its filtrate (through liquid) was taken, and nitrogen was quantified by the Keidel nitrogen method.

In addition, total nitrogen (TN) was measured by Keidel nitrogen method. For example, press to proceed.

60 ml of 0.05 M sodium citrate dihydrate solution heated to about 50° C. was added to 5 g of the sample (cheese), and homogenized at 8000 rpm for about 3 minutes with a rotary homogenizer. After washing the homogenizer with distilled water, 100 g of sample liquid was obtained. Take 2ml of the sample solution, and use the Keidel nitrogen method to quantify nitrogen. The obtained value is the amount of total nitrogen per 1 g of cheese.

The above-mentioned water-soluble nitrogen (WSN) was measured by Keidel nitrogen method. For example, proceed as follows.

60 ml of 0.05 M sodium citrate dihydrate solution heated to about 50° C. was added to 5 g of the sample (cheese), and homogenized at 8000 rpm for about 3 minutes with a rotary homogenizer. After washing the homogenizer with distilled water, 100 g of sample liquid was obtained. Stir it with a stirrer, and adjust the pH to 4.40±0.05 with 6 N hydrochloric acid solution. Filter with Toyo filter paper No.5A, take 2ml of the filtrate, and quantify nitrogen with the Keidel method. The obtained value is the amount of water-soluble nitrogen per 1 g of cheese.

The protease activity measurement described above is performed according to the Casein-Ferring method (Japan Food Additives Association), for example, as follows.

As the substrate for protease activity determination, 1.2 g of acid casein (ALACID720, Fonterra Corporation) was dissolved in 50 mM disodium hydrogen phosphate solution, adjusted to pH 7.0 with 1N hydrochloric acid, and filled with distilled water to 200 ml before use.

Add 5ml of matrix solution into a 20ml glass tube and incubate at 37°C. Inject 1ml of appropriately diluted enzyme solution into it to start the reaction. After 30 minutes, the reaction stop solution (0.44M trichloroacetic acid) was injected and left for 30 minutes to stop the enzyme reaction. The reaction solution was filtered with Toyo filter paper No. 2A, 5 ml of 0.55M sodium carbonate solution and 1 ml of 0.67N phenol reagent (Wako Pure Chemical Industries, Ltd.) were added to 2 ml of the filtrate, and the reaction was carried out at 37° C. for 30 minutes. Measure the absorbance of the chromogenic solution at 660nm. The amount of enzyme that produces amino acids equivalent to 10 μg of tyrosine in 1 ml of the reaction filtrate for 60 minutes was defined as 1 unit, and the activity of the enzyme per unit weight was calculated according to the following formula. The unit is unit/g.

unit/g＝(OD ₆₆₀ -OD ₀ )×117.6×(l/2)×(l/10)×N

OD ₆₆₀ : absorbance of the reaction filtrate

OD ₀ : absorbance of enzyme blank

117.6: The amount of tyrosine when the absorbance difference obtained from the tyrosine standard curve is 1

(l/2): Reaction filtrate volume

(l/10): unit conversion factor

N: Equivalent to the dilution factor of 1g or 1ml per sample

In addition, the measurement of the above-mentioned peptidase activity can be performed, for example, as follows.

As the substrate for the determination of peptidase activity, Lys-p-NA, a derivative of amino acid p-nitroanilide (hereinafter referred to as p-NA), was used.

Add 100 μl of 20 mM amino acid p-NA solution and 1.8 ml of 100 mM potassium phosphate buffer (pH 7.0, 37° C.) into a 5 ml glass tube, and incubate at 37° C. Inject 100 μl of an appropriately diluted enzyme solution to start the reaction. After 30 minutes, inject reaction stop solution (1.0ml of 30% (w/v) acetic acid to stop the enzyme reaction. Centrifuge at 10000rpm for 5 minutes after the reaction stops, and measure the absorbance of the supernatant at 410nm. The free p -NA has a maximum absorption at 410nm, the enzyme activity of free 1μmol p-NA in 1 minute is defined as 1 unit, and the enzyme activity per unit weight is calculated according to the following formula. The unit is unit/g.

unit/g＝(OD ₄₁₀ -OD ₀ )×1.13×(l/30)×(l/0.1)×N

OD ₄₁₀ : the absorbance of the enzyme reaction solution

OD ₀ : absorbance of enzyme blank

1.13: The amount of p-NA when the absorbance difference is 1 calculated from the standard curve

(1/30): reaction time

(l/0.1): Reaction volume

N: Equivalent to the dilution factor of 1g or 1ml per sample

In the present invention, the time when PTASN/TN×100 reaches 6 to 8.5% means that the flavor tendency of cheese corresponds to the ripening degree that can be confirmed by analysis. At this time, WSN/PTASN is 4.5 or less, because no bitter taste is produced. required.

When PTASN/TN×100 is less than 6%, it is not sufficient to grasp the ripening tendency (characteristic of proteolysis), and the actual ripening flavor cannot be reflected.

PTASN/TN×100 reaches 6-8.5%. This is because cheese usually takes 6-10 months at a curing temperature of 7-10°C, and 1.5-3 months at a storage temperature of 15°C. In order to promote ripening, cheese added with proteolytic enzymes derived from microorganisms as in the present invention can be judged to require 1 to 2 months for storage at 15°C and 2 weeks to 1 month for storage at 18 to 20°C.

As a feature of the addition of proteolytic enzymes derived from microorganisms, the protease activity must be 2000 unit/g or more, and the peptidase activity must be 160 unit/g or more. When the activity of protease is less than 2000unit/g, the decomposition of protein is slow, and the production amount of peptide is reduced. When the peptidase activity is lower than 160unit/g, the decomposition of peptides will be slow, and the bitter peptides will remain, and bitter cheese will be formed during the aging process.

As the microorganism-derived proteolytic enzymes, for example, "Blotea-Ze A'Amano'G" (trade name), "Brotea-Ze M'Amano'G" (trade name) manufactured by Amano Enzyme Co., Ltd., " "Umamizaim G" (trade name), "Pepchida-ZeR" (trade name), "Gultamine-Zedaiwa" (trade name), "BioFV" (trade name) manufactured by Kerry Food Ingredients, etc. From the viewpoint of improving the flavor of cheese, "Brotea-Ze A'Amano'G" (trade name), "Brotta-Ze M'Amano'G" (trade name) and "Uma Mizaim G" (trade name) are more preferable. , "BioFV" (trade name).

In the method for producing natural cheese of the present invention, cheese curd and/or cheese may not be sterilized. Any kind of lactic acid bacteria, protein crushed products, and proteolytic enzymes from microorganisms can be directly added to cheese curd or cheese, so enzymes from lactic acid bacteria or proteolytic enzymes from microorganisms, etc. The whey is drained together. That is, all or most of the actually added lactic acid bacteria, crushed lactic acid bacteria, proteolytic enzymes derived from microorganisms, etc. are preserved in the cheese curd and used, and the enzymes as the active ingredients are not lost (lost) or slightly loss.

In the method for producing natural cheese of the present invention, a pH adjuster may be added to the raw material milk (cheese milk) to form curd cheese. In the pH adjuster, the pH history (change over time) when lactic acid bacteria are added is passed. , Substances (food additives, etc.) that lower the pH of cheese milk are preferred, and specific examples include gluconolactone (GDL).

That is, when forming cheese curd, a pH adjuster may be used instead of a lactic acid bacterium starter, and a lactic acid bacterium starter may be used in combination with a pH adjuster.

In this case, the amount of gluconolactone added is 0.1 to 1% by weight, preferably 0.2 to 0.8% by weight, more preferably 0.3 to 0.6% by weight, based on the raw material milk (cheese milk). When the amount of gluconolactone added is greater than 1% by weight, when gluconolactone is added to cheese milk, the pH does not decrease slowly, so the moisture in the cheese curd is not fully discharged, and the moisture in the cheese (final product) increased.

The advantages of using a pH adjuster when forming a cheese curd are as follows.

(D-1) It eliminates the need for equipment investment for culture of lactic acid bacteria starter, management of operating conditions during culture, management of activity and number of lactic acid bacteria after the end of culture, etc., and reduces the burden of these.

(D-2) When using lactic acid bacteria starters to form cheese curds, the pH history needs to be known frequently, so it is necessary to choose lactic acid bacteria starters, but there is no need to worry about using pH regulators (GDL).

(D-3) When using lactic acid bacteria starter to form cheese curd, due to phage contamination, the pH of cheese milk decreases slowly, and cheese curd cannot be formed normally, but it is impossible to use pH regulators (GDL, etc.). That is, when lactic acid bacteria are added (supplemented) to cheese curds or cheese, there is no problem with bacteriophage contamination since pH drop does not occur. Here, the purpose of using lactic acid bacteria is to use the enzymes in the bacteria, and bacteriophage contamination may cause bacteriolysis. Of course, phage contamination may also have a positive effect.

(D-4) When using lactic acid bacteria starters to form cheese curds, it is necessary to select lactic acid bacteria starters considering the impact on the flavor and physical properties of cheese, but there is no such concern when using pH regulators (GDL, etc.). Therefore, attention is paid to the selection of post-addition (addition) of lactic acid bacteria to cheese curds or cheese, and the post-addition lactic acid bacteria tends to have a large influence on the formation of flavor. As a result, by using aged type natural cheese, flavor (scent) and physical properties can be individually adjusted arbitrarily, and a new natural cheese that has not been developed before has been developed.

(D-5) In order to ensure bacterial preservation of cheese, a pH regulator (GDL, etc.) may be used in combination with lactic acid bacteria when the flora must maintain the dominance of lactic acid bacteria at the stage of cheese curd formation.

Add lactic acid bacteria, crushed lactic acid bacteria, and proteolytic enzymes derived from microorganisms to cheese curds after whey has been drained (that is, cheese curds before aging) and/or cheese (that is, cheese at the beginning of aging). In the case of any one of these or any combination of these, it is effective to carry out the cheese curd and/or cheese after whey has been drained in a finely cut or pulverized state. That is, in the finely cut or pulverized state of the cheese curd and/or cheese after the whey has been discharged, the lactic acid bacteria culture solution or concentrated solution is sprayed, or the dry powder of lactic acid bacteria is added or mixed, or the lactic acid bacteria derived from microorganisms are added or mixed. proteolytic enzymes. When doing so, it is effective to thoroughly mix the whole of the cheese curd (that is, the cheese curd before aging) and/or the cheese (that is, the cheese at the initial stage of aging) by the enzyme derived from lactic acid bacteria or the proteolytic enzyme derived from microorganisms .

For example, in the production process of dry salty cheese, etc., after most of the whey is discharged, salt (powder) is added and mixed in the process of shredding cheese curds, and the dry powder of lactic acid bacteria can be added together with the salt, etc. without In particular, new processes are required.

On the other hand, when lactic acid bacteria are added to the aging cheese during the aging process, as in the case of cheese curds, when the cheese is shredded or pulverized, the lactic acid bacteria can be fully and effectively mixed in the whole cheese, but It is also possible to inject lactic acid bacteria liquid or powder into the cheese block with a needle or tube.

The method for producing natural cheese according to the present invention is particularly intended to be aged cheese, and examples of natural cheese produced by the method of the present invention include cheese, gouda, edam, emmental and cheese. Cheese, Parmesan, Camembert, Feta, etc.

The natural cheese of the present invention is an aged cheese produced by the above-mentioned production method, and its flavor characteristic is that the flavor is enhanced and the bitterness is suppressed.

Furthermore, the natural cheese of the present invention is obtained by, for example, adding any one of lactic acid bacteria, a crushed product of lactic acid bacteria cells, proteolytic enzymes derived from microorganisms, or any combination thereof to cheese curds after whey removal. At the beginning of aging, the soluble nitrogen content of the cheese after 90 days increases by 4% or more, preferably 5% or more, more preferably 6% or more compared with the beginning of aging.

In the present invention, the cheese soluble nitrogen content is the phosphotungstic acid (PTA) soluble nitrogen in the cheese total nitrogen, which is nitrogen contained in low molecular weight peptides or amino acids with a molecular weight of about 600 or less. These low-molecular-weight peptides or amino acids are produced by enzymatic decomposition of cheese curds or cheese proteins, and increase as the cheese matures.

The soluble nitrogen content is an index showing the progress of aging and is defined as follows.

Soluble nitrogen content [%] = PTA soluble nitrogen of cheese / total nitrogen of cheese × 100

Here, the PTA-soluble nitrogen or total nitrogen of the cheese can be measured by the Keidel method, and the above-mentioned steps can be used as an example of the pretreatment step in this case.

In addition, when evaluating the soluble nitrogen content, the soluble nitrogen content originating in the lactic acid bacteria culture liquid component etc. decreases, and it is defined as the soluble nitrogen content accompanying aging of cheese curd or cheese.

Fig. 1 is a graph showing the results of measuring the soluble nitrogen content of existing Gouda cheese by the above method. When the conventional Gouda cheese was used, the soluble nitrogen content from the beginning of aging to 90 days after aging increased by 2 to 3% compared with that at the beginning of aging.

In the natural cheese of the present invention manufactured by the production method of the present invention, it is assumed that the cheese soluble nitrogen content increases in the same tendency as before. This was confirmed in Examples 1 to 8 described later. The soluble nitrogen content of cheese after aging starts, from the viewpoint of improving the flavor, the soluble nitrogen content at 90 days after aging is increased by 4% or more, preferably 5% or more, more preferably 6% or more, compared with the beginning of aging.

According to the experiments of the inventors of the present invention, lactic acid bacteria are added to the cheese curd and/or cheese after the whey is discharged, so that the cell concentration reaches 10 ⁷ /g or more, or the cell crushing treatment of the lactic acid bacteria is added to make the cell When ripening after a concentration corresponding to 10 ⁵ /g or more, the soluble nitrogen content [%] after 3 months from the beginning of ripening increased by 4.0 to 8.5% compared with the beginning of ripening, which is good from the viewpoint of improving flavor. desired

In addition, when the cheese curd and/or cheese from which the whey was drained was aged by adding proteolytic enzymes derived from microorganisms, the soluble nitrogen content [%] at the time of 3 months after the start of aging increased by 6.0 compared with that at the beginning of aging. ~8.5%, which is desirable from an improved flavor point of view.

It is practical and typical to control (manage) the original aging (storage) temperature by adjusting the aging time and flavor of the cheese. That is, when you want to accelerate the aging of cheese, shorten the aging time, or enhance the flavor, you can increase the aging temperature; when you want to inhibit the aging of cheese, lengthen the aging time, or reduce the flavor, you can lower the aging temperature. Therefore, when you want to stop the ripening of cheese, you only need to set the ripening (preservation) temperature to ice temperature or freezing. However, when the aging of cheese is greatly accelerated, the aging temperature has an upper limit (about 15° C.) in view of the effects of bacterial growth, acidification reaction, and quality deterioration, so there is a limit to the effect of accelerating aging by aging temperature.

On the contrary, by controlling the curing temperature, according to the present invention, it can be freely adjusted to promote or inhibit the curing of cheese. Therefore, as described above, the method of adjusting the aging of cheese is applied not only to cheese curds before aging but also to cheese during aging, even in the middle of aging. In this regard, it is possible to predict the needs of various cheeses (products), manage them in the warehouse, or adjust the production volume, and calculate the order time (timing), which can relieve unstable and urgent business for cheese manufacturers or sellers . In particular, it is epoch-making that long-aging type cheese can be easily obtained in a short period of time.

Using the method for producing natural cheese of the present invention, for example, compared with the usual or existing necessary aging time, it is substantially shortened to 30-60%, preferably 30-70%, more preferably 30-80%, especially preferably 20-80%. 70%, most preferably 20-80%.

Hereinafter, examples related to the present invention will be given and described, but the present invention is not limited thereto.

(Preparation of cheese curd for cheese cheese)

The amount of milk used is 100kg, and cheese curds are first prepared. Raw material milk (milk for cheese) is used, the ratio of protein/fat is adjusted to 0.8, and it is heat-sterilized at 63° C. for 30 minutes. After adding 0.03 weight% of potassium chloride to this sterilized milk for cheese, 1.2 weight% of lactic acid bacteria fermentation products, and 0.01 weight% of rennet were added. After confirming the coagulation, it is cut and prepared to produce whey-expelled cheese. The lactic acid bacteria starter contains mixed bacteria of Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris. The cheese curds from which the whey has been discharged are pressure-molded, cut into squares of about 2 cm, and divided into 3 equal parts by weight to make cheese curds 1, 2, and 3.

Example 1

(Preparation of Cheese Cheese)

Add salt to cheese curd 1 to make the salt content of cheese curd reach 1.8% by weight, then add 2% by weight of lactic acid bacteria concentrate, mix and vacuum pack. This lactic acid bacteria concentrate contains only Lactococcus crispatus 10 ¹⁰ /g. After the lactic acid bacteria concentrate is neutralized and cultivated, the lactic acid bacteria are concentrated by centrifugation to complete the preparation. Then, the vacuum-packed cheese curd was matured at 10°C.

Example 2

(Preparation of Cheese Cheese)

Add salt to the cheese curd 2 to make the salt content of the cheese curd reach 1.8% by weight, then add 2% by weight of the lactic acid bacteria concentrate processed by crushing the bacteria, mix and vacuum pack. Before the crushing treatment of the bacterium, the lactic acid bacteria concentrate only contains 10 ¹⁰ Lactococcus crispatus/g. After the bacterial cell crushing treatment, it contained 10 ⁷ cells/g. After the lactic acid bacteria concentrate is neutralized and cultivated, the lactic acid bacteria are concentrated by centrifugation to complete the preparation. In addition, a high-pressure homogenizer (operating pressure: 140 MPa) was used during the crushing treatment of the bacteria. Then, the vacuum-packed cheese curd was matured at 10°C.

Comparative example 1

(Preparation of Cheese Cheese)

Only salt was added to the cheese curd 3 to make the salt content of the cheese curd 1.8% by weight, and vacuum packaging was performed after mixing. Then, the vacuum-packed cheese curd was matured at 10°C.

Regarding the cheeses of Example 1, Example 2, and Comparative Example 1, the relationship between the aging time and the soluble nitrogen content is shown in FIG. 2 . In Comparative Example 1, the soluble nitrogen content of the cheese after 90 days of maturation was about 3.5%, which was lower than 4%, while in Examples 1 and 2, the soluble nitrogen contents after 90 days were 5.3% and 7.4%, respectively. Any one is also above 5%. In addition, in Comparative Example 1, the aging time for the soluble nitrogen content of the cheese to reach about 3.5% was about 90 days, while in Example 1 and Example 2, it was about 45 days and about 30 days, respectively. At this time, in Example 1 and Example 2, compared with Comparative Example 1, the curing time is shortened to about 50% (1/2) and about 33% (1/3). That is, in Example 1 and Example 2, aging was accelerated compared with Comparative Example 1, and the actual aging time was shortened.

The relationship between the aging time and the enzyme activity of the cheeses of Example 1, Example 2, and Comparative Example 1 is shown in FIG. 3 . Enzyme activity increases in order according to Example 2>Example 1>Comparative Example 1, and the order of enzyme activity is high, indicating that the generation of soluble nitrogen is fast, and the aging is carried out rapidly. At this time, when the aging time was 0 days (curing start), Example 1 was 0.17 ABS/h, Example 2 was 0.62 ABS/h, and Comparative Example 1 was 0.02 ABS/h. When Example 1 was compared with Comparative Example 1, the enzyme activity was increased to about 10 times by adding live bacteria of lactic acid bacteria to cheese curd. When Example 2 was compared with Comparative Example 1, the enzyme activity was increased to about 30 times by the bacterial cell disruption treatment. In addition, when the curing time is 90 days, the 0.53ABS/h in Example 1, 0.78ABS/h in Example 2, and 0.04ABS/h in Comparative Example 1. When Example 1 was compared with Comparative Example 1, the enzyme activity was increased to about 10 times by adding live bacteria of lactic acid bacteria to cheese curd. When Example 2 was compared with Comparative Example 1, the enzyme activity was increased to about 20 times by the bacterial cell disruption treatment.

In the present invention, the above enzymatic activity is a value measured by the following method.

(E-1) Add 80 ml of extraction buffer (37° C.) to 20 g of lactic acid bacteria concentrate or crushed solution, and use a homogenizer (manufactured by IKA, model: ULTRA-TURRAX T25) to process for 1 minute at 9500 rpm . Here, the extraction buffer is an aqueous solution containing potassium phosphate buffer (50 mM, pH=7 (37° C.)), sucrose (30 w/v%), and sodium chloride (150 mM).

(E-2) Take 30 g of the aqueous solution of (E-1) above, and perform centrifugation at 4° C., 8000 g, for 10 minutes.

(E-3) The supernatant was filtered with filter paper (TOYO No. 2), and the filtrate (passed liquid) was used as an extract of lactic acid bacteria.

(E-4) When measuring the enzyme activity, the amino acid lysine (hereinafter referred to as Lys) and p-nitroanilide (hereinafter referred to as p-NA) derivative Lys-p-NA (Sigma Corporation ) as the matrix. Put 100 μl of Lys-p-NA solution (20 mM) and 1.8 ml of potassium phosphate buffer (100 mM, pH=7 (37°C)) into a small test tube (5 ml), and incubate at 37°C.

(E-5) Take 100 μl of the extract of lactic acid bacteria and put it into the above-mentioned (E-4) small test tube to start the reaction.

(E-6) After 0, 2, 4, and 6 hours from the start of the reaction, 1.0 ml of acetic acid (30w/v%) was put into the above-mentioned small test tube (E-5), and the reaction was stopped at each time.

(E-7) The p-NA freed by the above reaction has a maximum absorption at a wavelength of 410 nm, and the liquid of (E-6) above is centrifuged at 10,000 rpm for 5 minutes.

(E-8) The absorbance of the supernatant was measured with a spectrophotometer (manufactured by Shimadzu Corporation, model: UV-1200, wavelength 410 nm).

(E-9) The results of each reaction time measurement were plotted with the horizontal axis representing the reaction time (h) and the vertical axis representing the absorbance (ABS) to obtain an approximate quadratic curve formula. Then, using this quadratic curve, the first-order coefficient was defined as the enzyme activity (ABS/h).

The flavors of the cheeses of Example 1, Example 2, and Comparative Example 1 were compared by sensory evaluation. The result of the sensory evaluation after 90 days from the start of aging is that, compared with Comparative Example 1, Example 1 or Example 2 has a strong overall aroma, no bitterness, and a strong aroma.

(preparation of cheese curds for dry and salty Gouda cheese)

Dry salty Gouda cheese was produced on a scale of 100 kg of milk. First, cheese curd was prepared. Raw material milk (milk for cheese) is used, the ratio of protein/fat is adjusted to 1.0, and it is heat-sterilized at 63° C. for 30 minutes. After adding 0.03 weight% of calcium chloride to this sterilized cheese milk, 1 weight% of lactic acid bacteria fermented product, and 0.01 weight% of rennet were added. After confirming the coagulation, the cheese is cut and prepared to produce cheese from which the whey has been expelled. The lactic acid bacteria starter contains mixed bacteria of Lactococcus lactis subsp. lactis, L. lactis subsp. lactis biovar diacetilactis and L. lactis subsp. cremoris. After the cheese curds discharged from the whey are press-molded, they are cut into about 2 cm squares and divided into 5 equal parts by weight to make cheese curds 4, 5, 6, 7, and 8.

Example 3

(Preparation of dry and salty Gouda cheese)

Add salt to the cheese curd 4 to make the salt content of the cheese curd reach 1.7% by weight, then add 2% by weight of lactic acid bacteria concentrate, mix and vacuum pack. The lactic acid bacteria concentrate only contains Lactobucillus he1veticus 10 ¹⁰ /g. After the lactic acid bacteria concentrate is neutralized and cultivated, the lactic acid bacteria are concentrated by centrifugation to complete the preparation. Then, the vacuum-packed cheese curd was matured at 10°C.

Example 4

(Preparation of dry and salty Gouda cheese)

Add salt to the cheese curd 5, so that the salt content of the cheese curd reaches 1.7% by weight, and then add 2% by weight of the lactic acid bacteria concentrate processed by crushing the bacteria cells, mix and vacuum pack. Before the thalline crushing treatment, the lactic acid bacteria concentrate only contains ¹⁰ pcs/g of Lactobucil lus he1veticus10. After the bacterial cell crushing treatment, it contained 10 ⁷ cells/g. After the lactic acid bacteria concentrate is neutralized and cultivated, the lactic acid bacteria are concentrated by centrifugation to complete the preparation. In addition, a multi-hollow particle electric shock generator (manufactured by Yasui Instrument Co., Ltd., hollow particle diameter: 0.3 mm) was used for the crushing treatment of the bacterial cells. Then, the vacuum-packed cheese curd was matured at 10°C.

Comparative example 2

(Preparation of dry and salty Gouda cheese)

Only salt was added to the cheese curd 6 so that the salt content of the cheese curd reached 1.7% by weight, and vacuum packaging was performed after mixing. Then, the vacuum-packed cheese curd was matured at 10°C.

Regarding the cheeses of Example 3, Example 4, and Comparative Example 2, the relationship between the aging time and the soluble nitrogen content is shown in FIG. 4 . In Comparative Example 2, the soluble nitrogen content of the cheese after 90 days of maturation was about 2.8%, lower than 3%, while in Examples 3 and 4, the soluble nitrogen contents after 90 days were 4.3% and 6.8%, respectively. Any kind is also above 4%. In addition, in Comparative Example 2, the aging time for the soluble nitrogen content of the cheese to reach about 2.5% was about 60 days, while in Examples 3 and 4, it was about 30 days and about 20 days, respectively. At this time, in Example 3 and Example 4, compared with Comparative Example 2, the curing time is shortened to about 50% (1/2) and about 33% (1/3). That is, in Example 3 and Example 4, aging was accelerated compared with Comparative Example 2, and the actual aging time was shortened.

Regarding the cheeses of Example 3, Example 4, and Comparative Example 2, the flavors were compared by the senses. As a result of the sensory evaluation after 90 days from the start of aging, compared with Comparative Example 2, Example 3 and Example 4 had a strong overall aroma, no bitterness, and a strong aroma.

Example 5

(Preparation of dry and salty Gouda cheese)

Add salt to the cheese curd 7 to make the salt content of the cheese curd reach 1.7% by weight, then add 2% by weight of lactic acid bacteria concentrate, mix and vacuum pack. The lactic acid bacteria concentrate contains 1010 mixed bacteria of ^{Lactobucillus} lactis subsp. lactis, L. lactis subsp. lactis biovar diacetilactis and L. lactis subsp. cremoris/g. After the lactic acid bacteria concentrate is neutralized and cultivated, the lactic acid bacteria are concentrated by centrifugation to complete the preparation. In addition, this vacuum-packed cheese curd was aged at 10°C.

Example 6

(Preparation of dry and salty Gouda cheese)

Add salt to the cheese curd 8, so that the salt content of the cheese curd reaches 1.7% by weight, and then add 2% by weight of the lactic acid bacteria concentrate processed by crushing the bacteria cells, mix and vacuum pack. Before the cell crushing treatment, the lactic acid bacteria concentrate contained 10 ¹⁰ mixed bacteria of Lactococcus lactis subsp.lactis, L.lactis subsp.lactis biovardiacetilactis, L.lactis subsp.cremoris/g, and after the cell crushing treatment contained 10 ⁷ pcs/g. After the lactic acid bacteria concentrate is neutralized and cultured, the lactic acid bacteria are centrifuged and concentrated to complete the preparation. In addition, a multi-hollow particle electric shock generator (manufactured by Yasui Instrument Co., Ltd., multi-hollow particle diameter: 0.3 mm) was used for the crushing treatment of the bacterial cells. Then, the vacuum-packed cheese curd was matured at 10°C.

Regarding the cheeses of Example 5, Example 6, and Comparative Example 2, the relationship between the aging time and the soluble nitrogen content is shown in FIG. 5 . In Comparative Example 2, the soluble nitrogen content of the cheese after 90 days of maturation was about 2.8%, lower than 3%, while in Examples 5 and 6, the soluble nitrogen contents after 90 days were 3.6% and 5.2%, respectively. Any kind is also above 3.5%. In addition, in Comparative Example 2, the aging time for the cheese soluble nitrogen content to reach about 2.5% was about 60 days, while in Example 5 or Example 6, it was about 50 days and about 40 days, respectively. At this time, in Example 5 or Example 6, compared with Comparative Example 2, the curing time is shortened to about 83% (5/6) and about 66% (2/3). That is, in Example 5 or Example 6, aging was accelerated compared with Comparative Example 2, and the actual aging time was shortened.

Regarding the cheeses of Example 5, Example 6, and Comparative Example 2, flavors were compared by sensory perception. As a result of the sensory evaluation after 90 days from the start of aging, compared with Comparative Example 2, Example 5 or Example 6 has a stronger overall aroma, no bitterness, and a stronger aroma.

(Preparation of Gouda Cheese with pH Regulator)

Using gluconolactone (GDL, manufactured by Fuso Chemical Industry Co., Ltd.), cheese curd was produced on a scale of 10 kg of milk. Raw material milk (milk for cheese) is used, the ratio of protein/fat is adjusted to 0.8, and it is heat-sterilized at 63° C. for 30 minutes. After adding 0.03 weight% of potassium chloride to this sterilized milk for cheese, 0.5 weight% of gluconolactone and 0.01 weight% of rennet were added. After confirming the coagulation, it is cut and prepared to produce curd cheese from which the whey has been expelled. At the time of cheese curd production, storage temperature is about 30 degreeC. When producing cheese curd, gluconolactone and rennet are added, and the whey is discharged when the pH reaches 5.40. After the cheese curds discharged from the whey are press-molded, they are cut into squares of about 2 cm in size and divided into 2 equal parts by weight to make cheese curds 9 and 10 .

Example 7

(Preparation of Gouda Cheese)

Add salt to the cheese curd 9 to make the salt content of the cheese curd reach 1.8% by weight, then add 2% by weight of lactic acid bacteria concentrate, mix and vacuum pack. This lactic acid bacteria concentrate only contains Lactobacillus crispatus 10 ¹⁰ /g. The lactic acid bacteria concentrate is prepared by centrifuging and concentrating the lactic acid bacteria after being neutralized and cultivated. Then, the vacuum-packed cheese curd was matured at 10°C.

Comparative example 3

(Preparation of Gouda Cheese)

Only salt is added to the cheese curd 10 so that the salt content of the cheese curd becomes 1.8% by weight, and vacuum packaging is performed after mixing. Then, the vacuum-packed cheese curd was matured at 10°C.

About the cheese of Example 7 and Comparative Example 3, the soluble nitrogen content after aging was measured. In Comparative Example 3, the soluble nitrogen content after 90 days from the start of aging was about 1.9%, while in Example 7, the soluble nitrogen content after 90 days was about 4.0%. The soluble nitrogen content of cheese increased after 90 days.

Regarding the cheeses of Example 7 and Comparative Example 3, the flavors were compared by the senses. As a result of the sensory evaluation after 90 days from the start of aging, the cheese flavor of Comparative Example 3 was lacking, but in Example 7, the aroma or aroma of cheese was strong, and it was very tasty.

Example 8

Prepare the curd of soda cheese according to the usual method (add lactic acid bacteria starter and crude rennet to the sterilized raw milk, cut and prepare after confirming the coagulation, and discharge the whey). After eggplantization, grinding, adding salt, adding microbial decomposing enzymes from protein, mixing evenly with curd milk, forming, packaging, and aging at 10°C.

The proteolytic enzyme derived from microorganisms was BIO FV manufactured by Kerry Food Ingredients, the peptidase activity was 180u/g, and the amount added to the curd was 100ppm.

The trial cheese was stored at 7°C for 60 days. At this time, PTASN/TN×100=6.86%, and WSN/PTASN=1.86.

The flavor of this preserved product was non-bitter and had a good cheese flavor.

Possibility of industrial use

According to the present invention, there is provided a method for producing natural cheese that effectively controls and adjusts aging time and flavor (especially aroma) through simple operations. In addition, it provides a method for producing aged-type natural cheese that promotes ripening, enhances flavor, and suppresses bitterness, which is particularly practical in mass production and continuous production.

Claims (10)

1. A method for producing aged natural cheese, which is characterized in that lactic acid bacteria are added to the cheese curd and/or cheese from which the whey has been discharged so that the bacterial cell concentration reaches 10 ⁷ /g or more, and then aging is carried out, and aging starts The aging time until the final soluble nitrogen content (%) reaches a predetermined value is shortened to 20 to 80% compared with the case where the lactic acid bacteria are not added, and the soluble nitrogen content [%] at the time of 3 months after the initiation of aging is 3.5% or more. 2. The method according to claim 1, wherein the lactic acid bacteria are live bacteria. 3. A method for producing aged natural cheese, characterized in that a crushed lactic acid bacterium is added to the cheese curd and/or cheese from which the whey has been discharged so that the concentration of the bacterium corresponds to 10 ⁵ cells/g After the above, aging is carried out, and the aging time until the soluble nitrogen content (%) reaches a predetermined value after the initiation of aging is shortened to 20 to 80% compared with the case of not adding the crushed bacterium of the lactic acid bacteria, and aging begins. The soluble nitrogen content [%] after 3 months passed was 3.5% or more. 4 . The method for producing aged natural cheese according to claim 3 , wherein the bacterium crushing treatment is a homogenization treatment under an operating pressure of 100 MPa or more. 5 . 5. The method for producing aged natural cheese according to any one of claims 1 to 4, wherein lactic acid bacteria concentrated by any method of membrane separation, centrifugal separation, and vacuum evaporation are used. 6. The method for producing aged natural cheese according to any one of claims 1 to 4, wherein lactic acid bacteria dried by any one of freeze-drying, reduced-pressure spray-drying, and spray-drying are used. . 7. The method for producing aged natural cheese according to any one of claims 1 to 6, wherein neutralized cultured lactic acid bacteria are used. 8. A method for producing matured natural cheese, which is characterized in that, after adding proteolytic enzymes from microorganisms to the cheese curd and/or cheese from which the whey has been discharged, aging, phosphotungsten acid soluble nitrogen (Phospho Tangsten Acid Soluble Nitrogen) (PTASN) to total nitrogen (Total Nitrogen (TN)) ratio (PTASN/TN × 100) reached a predetermined value after the start of aging aging time compared to the situation without adding the proteolytic enzyme derived from microorganisms was shortened to 20 ～80%, the soluble nitrogen content [%] at the time of 3 months after the start of aging is increased by 6.0～8.5% compared with the beginning of aging, wherein the protease activity of the proteolytic enzyme derived from microorganisms is 2000unit/ g above, and the peptidase activity is above 160unit/g. 9. according to the manufacture method of the aging type natural cheese described in claim 8, it is characterized in that, add the phosphotungsten acid soluble nitrogen (Phospho Tangsten Acid SolubleNitrogen (PTASN)) of the cheese that carries out aging after adding the proteolytic enzyme from microorganism to total When the nitrogen (Total Nitrogen (TN)) reaches 6.0-8.5% (PTASN/TN×100), the ratio of Water Soluble Nitrogen (WSN) to PTASN (WSN/PTASN) is below 4.5. 10. The method for producing aged natural cheese according to any one of claims 1 to 9, wherein the cheese curd and/or cheese from which the whey has been drained are not sterilized.