KR20160051084A - composition for increasing the storage stability of culture product of lactic acid bacteria - Google Patents

Abstract

The present invention relates to a composition for enhancing storage stability of a culture of a lactic acid bacterium, comprising a cyclosporin, a method for increasing the storage stability of a lactic acid bacterium culture using a lactic acid bacterium and a milk-containing raw material using a scicos, As to the increased dairy product, the lactic acid bacterium culture product or dairy product according to the present invention has an effect of maintaining an appropriate range of pH and pH during the storage period and preventing the false-positives.

Description

[0001] The present invention relates to a composition for enhancing storage stability of cultured lactic acid bacteria,

The present invention relates to a composition for enhancing storage stability of a culture of a lactic acid bacterium, comprising a cyclosporin, a method for increasing the storage stability of a lactic acid bacterium culture using a lactic acid bacterium and a milk-containing raw material using a scicos, As to the increased dairy product, the lactic acid bacterium culture product or dairy product according to the present invention has an effect of maintaining an appropriate range of pH and pH during the storage period and preventing the false-positives.

Recently, as interest in health has increased, studies have been conducted on foods containing lactic acid bacteria known to secrete various useful physiologically active substances in the human body. Lactic acid bacteria prevent fermentation of lactic acid by fermenting food, secrete antimicrobial substances such as bacteriocin to inhibit food poisoning bacteria, lower the pH of human intestinal pH and inhibit the growth of intestinal bacteria. Lt; / RTI >

Among the lactic acid bacteria fermented dairy products, there are cheese, buttermilk, yogurt, etc. Recently, a variety of functional yogurt has been widely marketed. Lactobacillus, lactobacillus, and lactococcus lactic acid bacteria are inoculated alone or as a mixed starter, Fermented dairy products having high functionality are being released.

However, since fermented dairy products using lactic acid bacteria are required to remove unique fermentation wounds and improve their aesthetics, lactic acid bacteria are included in the form of live bacteria, so that there is a problem that the products are easily deteriorated during storage or distribution. The fermented dairy products are then circulated and stored in a refrigerated or frozen state. Therefore, it is very important to maintain various properties (pH, acidity, etc.), taste, shape and number of lactic acid bacteria in the fermented dairy product during the storage period.

The method of adding pectin in the production of a lactic acid bacterium culture solution is widely used in order to prevent phase separation or superficial phase, which is one of the physical properties of fermented dairy products. However, since pectin is mainly used but pectin has low solubility, it is difficult to use more than a certain amount.

In addition, Korean Unexamined Patent Publication No. 2014-0102888 discloses a method for efficiently storing cultured lactic acid bacterium culture medium while maintaining the viable cell count for a long period of time. In the culture medium of lactic acid bacteria, NaHCO ₃ , Al (OH) ₃ , Mg (OH) ₂ , CaCO ₃ and NaAl OH) ₂ CO < ₃ > is added and stored at a refrigeration temperature of 3 to 5 [deg.] C.

There is still a need for fermented dairy products having high storage stability by controlling various physical properties (pH, acidity, etc.), taste, shape, and viable cell count of cultured products using lactic acid bacteria and fermented dairy products within a certain range during distribution and storage .

An example of the present invention is to provide a composition for enhancing the storage stability of a cultured Lactobacillus acid bacteria comprising a cyclosporin.

Another example of the present invention is to provide a method for increasing the storage stability of a lactic acid bacteria culture using a lactic acid bacterium and a milk-containing raw material by using a scicos.

In addition, one example of the present invention is to provide a dairy product having enhanced storage stability using the Saikos.

The present invention relates to a composition for enhancing storage stability of a culture of a lactic acid bacterium, comprising a cyclosporin, a method for increasing the storage stability of a lactic acid bacterium culture using a lactic acid bacterium and a milk-containing raw material using a scicos, As to the increased dairy product, the lactic acid bacterium culture product or dairy product according to the present invention has an effect of maintaining an appropriate range of pH and pH during the storage period and preventing the false-positives.

The present invention relates to a composition for enhancing storage stability of a cultured lactic acid bacterium, which comprises psicose.

Another embodiment of the present invention is directed to a method for increasing the storage stability of a lactic acid bacterium culture product using lactic acid bacteria and a milk-containing raw material, using the Saicos.

A further example of the present invention relates to a fermented dairy product comprising a composition for enhancing storage stability of a culture of a lactic acid bacterium containing a cyclus and a culture of a lactic acid bacterium produced by culturing a lactic acid bacterium in a milk-containing raw material.

The composition for enhancing storage stability of a culture of a lactic acid bacterium containing a cycloserone according to the present invention has an advantage that the physical properties, taste and properties of a lactic acid bacterium cultured product or a fermented dairy product can be maintained within a certain range during a storage period. The lactic acid bacterium culture product or the fermented milk product according to the present invention may further comprise an additive selected from the group consisting of modified starch and pectin. The addition of the modified starch also has the effect of imparting a soft texture of the fermented milk. The pectin used in the lactic acid bacteria fermented product or the fermented milk product for the purpose of preventing the layer separation or the deterioration phase has a disadvantage that the solubility thereof is poor and the composition for enhancing the storage stability with the psicose according to the present invention, alone or in combination with pectin, Can be further improved.

Hereinafter, the present invention will be described in more detail.

The composition for enhancing storage stability of a lactic acid bacterium culture product or fermented milk product according to the present invention includes a scicos known as a rare saccharide.

The composition containing the psicose according to the present invention increases the storage stability of the lactic acid bacteria culture product or the fermented milk product, that is, the storage stability of the lactic acid bacteria cultured product or the fermented milk product during storage is improved by various properties (pH, It is possible to control the number to a certain range. In a specific example of the present invention, the storage stability of the lactic acid bacteria culture or the fermented milk product includes controlling the pH range, controlling the acidity range, controlling the range of the number of lactic acid bacteria living bacteria, and controlling the phase separation (or isomerization phase) to a certain range or less.

Lactic acid bacteria cultures or fermented dairy products shall be kept at constant quality during storage or distribution, and the acidity range is required to meet the requirement of 0.5% or more of the Korean industrial standard, for example, in the range of 0.7 to 1.2% , and the pH range may be maintained in the range of 3.5 to 4.6 to achieve storage stability.

In addition, the composition containing the psicose may prevent phase separation or phase separation of lactic acid bacteria cultured product or fermented milk product during the storage period. For example, when the volume of the liquid generated per volume of whole lactic acid bacteria culture during storage is in percentage The indicated completion ratio may be 5 v / v% or less, for example, 4 v / v% or less, and 3 v / v% or less, and most preferably 0.

In general, psicose is almost similar in intensity and type of sweetness when compared to fructose, an epimer of d-fructose. However, unlike fructose, it is rarely metabolized when absorbed into the body, and has a function of inhibiting glucose absorption by inhibiting glucose absorption. Thus, it can be used for foods and beverages for diabetic patients or for food and drink for recipients. And can inhibit the accumulation of abdominal fat, so it can be used for various functional foods such as health food.

The sauces used in the present invention are not particularly limited and can be used as long as they can achieve storage stability of the fermented lactic acid bacteria. In a specific example, the Saicos may be powdery or liquid and may be a Saicos with a purity of at least 90%, for example 99%, or may further comprise components such as Saicos and other monosaccharides, disaccharides, and oligosaccharides Mixed sugar forms may also be presented, for example, in the form of syrups of mixed sugars.

In the present invention, the saccharose may be added before, during or after cultivation of the lactic acid bacteria when the lactic acid bacteria culture solution or fermented milk product is used for improving the storage stability, and the addition time is not particularly limited. However, the fermented milk product may be added to the raw material for fermented milk product in consideration of the convenience of the addition process or may be added after the fermented milk product is prepared.

The solid content of the cyclosporine according to the present invention is 2 to 55 parts by weight, preferably 2 to 20 parts by weight, more preferably 2 to 15 parts by weight, relative to 100 parts by weight of the total solid content of the composition for enhancing storage stability, , And more preferably 2 to 10 parts by weight. The content of the scissors contained in the composition according to the present invention can be appropriately adjusted within the above range in consideration of the appropriate degree of sweetness of the composition and the storage stability intended for the purpose of addition of the scissors.

The saccharose contained in the composition for enhancing storage stability according to the present invention may be a solution prepared by dissolving 100% psicose in powder or various concentrations. In addition, the above-mentioned synuclein may be a mixed saccharide containing a saccharose. Examples of saccharose saccharides include a saccharose, and further may include at least one selected from the group consisting of fructose, glucose, and oligosaccharides. Specific examples of the saccharide-containing mixed sugar include 2 to 55 parts by weight of psicose, 30 to 80 parts by weight of fructose, 2 to 60 parts by weight of glucose, and 0 to 15 parts by weight of oligosaccharide based on 100 parts by weight of the total solid content of the mixed sugar And may not include oligosaccharides. Saikos, fructose and glucose are preferably both D-isomer.

Saikosu can be carried out by chemical synthesis, or by biological methods using a cyclic epimerase, preferably by a biological method. Thus, the above-mentioned psicose includes at least one selected from the group consisting of psicose epimerase, cells of the strain producing the enzyme, culture of the strain, lysate of the strain, and extracts of the lysate or the culture The composition for producing psicose may be one prepared by reacting with a fructose-containing raw material.

In one embodiment of the present invention, a method for producing a scikos according to a biological method includes a method of culturing a strain producing a < RTI ID = 0.0 > cytokine < / RTI > epimerizing enzyme or a recombinant strain into which a gene encoding a scikos epimerase has been introduced, The course epimerase can be produced by reacting with a fructose-containing raw material. The above-mentioned cyclic epimerase can be carried out as a solid phase reaction using a liquid phase reaction or an immobilized enzyme.

Alternatively, a strain producing a cytomegalovirus or a recombinant strain into which a gene encoding a cytomegalovirus is introduced is obtained, and the culture of the strain, the culture of the strain, the lysate of the strain, and the lysate of the lysate or the culture Extract of the present invention can be produced by reacting a composition for producing a scorch, which comprises at least one member selected from the group consisting of a fructose-containing raw material, and a fructose-containing raw material. In the case of producing the cytosine using bacterial strains producing the Escherichia epimerase, the reaction can be carried out in the solid phase reaction using a liquid reaction or immobilized cells.

In a specific example of the present invention, as a strain producing a psicose epimerase, it may be a strain capable of producing a psicose epimerase at a high yield while having high stability. The recombinant strain may be a variety of host cells such as Escherichia coli, Bacillus sp., Salmonella sp., And Corynebacterium sp., But may be preferably a strain of K. niger bacteria, GRAS strain, It can be glutaricum.

In the case of using a recombinant strain, the gene encoding an enzyme derived from various strains can be used as a cytomegalic epimerase. For example, an enzyme derived from trifoneme primitia described in Korean Patent Publication No. 2014-0021974, Korean Patent Publication No. 2014-0080282 Or an enzyme derived from Clostridials dans according to Korean Patent No. 10-1318422, and may also be an enzyme derived from enciphered Halsen. In a specific example, the cytosine epimerase according to the present invention may be an enzyme derived from a clostridial synthase, for example, having an amino acid sequence of SEQ ID NO: 7, and a nucleic acid sequence of SEQ ID NO: 8 or SEQ ID NO: 9. The nucleic acid sequence of SEQ ID NO: 8 is an E. coli optimized nucleic acid sequence and the SEQ ID NO: 9 is a nucleic acid sequence modified appropriately to Corynebacterium.

In the production of the recombinant strain according to an embodiment of the present invention, the expression of the enzyme may be regulated using a regulatory sequence located on the upper side of the nucleic acid sequence encoding the above-mentioned cyclic epimerase, and the regulatory sequence is essentially a transcription promoter And may further include ribosome binding regions and / or spacer sequences, and the like. The elements constituting the regulatory sequence may be directly linked or linked by including one or more linkers of the nucleic acid sequence having 1 to 100 bases, for example, 5 to 80 bases.

In one embodiment, the transcriptional promoter may be a nucleic acid molecule that expresses a nucleic acid sequence encoding a cyclic epimerase in a Corynebacterium sp. Strain, but may be a tac1, tac2, trc, or sod promoter. The sod promoter is derived from Corynebacterium glutaricum, and preferably comprises the nucleotide sequence of SEQ ID NO: 1 as a core region. The trc promoter is an Escherichia coli -derived promoter produced by a combination of the trp promoter and the lac UV5 promoter. The Tac1 promoter is an Escherichia coli-derived promoter, which is produced by a combination of the trp promoter and the lac UV5 promoter. The Tac2 promoter is an Escherichia coli-derived promoter, which is prepared by a combination of the trp promoter and the lac UV5 promoter, and is optimized by modifying the sequence of the Tac1 promoter.

The ribosome binding region and the spacer may be chemically connected directly or indirectly via a linker nucleic acid sequence in between. In an embodiment of the present invention, the ribosome binding region and the spacer sequence may include one oligonucleotide sequentially linked from 5 'to 3'. The nucleotide sequences of the promoter sequence, the ribosome binding region and the spacer sequence according to an embodiment of the present invention are shown in Table 1 below. The boxed portions in Table 1 indicate the ribosome binding region, the spacer sequence, the linker sequence, and the like among the regulatory sequences.

order
number Sequences (5 '-> 3') denomination One aagcgcc tcatcagcgg taaccatca cgggttcgggt gcgaaaaacc atgccataac aggaatgttc ctttcgaaaa ttgaggaagc cttatgccct tcaaccctac ttagctgcca attattccgg gcttgtgacc cgctacccgataaataggtc ggctgaaaaa tttcgttgca atatcaacaa aaaggcctat cattgggaggtgtcgcacca agtacttttg cgaagcgcca tctgacggat tttcaaaaga tgtatatgct cggtgcggaa acctac
gaaagga ttttttaccc atggctg tatacgaact cccagaactc gactacgcat acgac
gaaagga ttacaaa Sod promoter
2 tgacaattaatcatcggctcgtatattgt gtggaattgtgagcggataacaatttcacacaggaaacagaattcccggg gaaagga ttacaaa tac1 promoter
3 Tgacaattaatcatccggctcgtataatgt taacaatttgtggaattgtgagcggacacacaggaaacagaccatggaattcgagctcggtacccggg gaaagga ttacaaa Tac2 promoter
4 Tgacaattaatcatcggcctcgtataatgt trc promoter
5 Gaaagga Ribosome binding region 6 Ttacaaa Spacer sequence

It is preferable that the cyclic epimerase according to the present invention is excellent in enzyme activity and thermal stability. Accordingly, in the embodiment of the present invention, the combination of the transcriptional promoter or the regulatory sequence with the gene encoding the cyclic epimerase is important , The tac1, tac2, trc, and sod promoters used in the present invention can provide more than adequate titers of protein expression, and when the sod promoter is used, folding of the protein is robust, It is more preferable to obtain a result which is high.

The method of producing a psicos using a recombinant strain can be carried out according to the method described in Korean Patent Publication Nos. 2014-0021974, 2014-0080282 and Korean Patent No. 10-1318422, but is not particularly limited. In the above-mentioned method of producing a scicos, the concentration of fructose used as a substrate may be 40 to 75% (w / v), for example, 50 to 75% (w / v) have. When the concentration of fructose is lower than the above range, economical efficiency is lowered. When the concentration of fructose is higher than the above range, fructose is not dissolved well, so that the concentration of fructose is preferably within the above range. The fructose may be used in the form of a solution dissolved in a buffer solution or water (e.g., distilled water).

The composition for enhancing storage stability according to the present invention can be applied to a culture solution of a lactic acid bacterium, for example, a culture solution of a lactic acid bacterium obtained by inoculating and culturing a lactic acid bacterium into various milk-containing raw materials or various fermented dairy products containing the same.

The milk-containing raw material may include at least one selected from the group consisting of raw milk, low fat milk, non-fat milk, reduced milk, reduced low fat milk and skim milk powder, and if the culture is suitably performed, Not limited.

The lactic acid bacterium for producing the lactic acid bacterium according to the present invention is not particularly limited, and various types of lactic acid bacteria such as Lactobacillus sp., Bifidobacterium sp., Streptococcus sp. Lactic acid bacteria can be used. Can be used as the above mixture. The lactic acid bacterium according to an embodiment of the present invention may be selected from the group consisting of Lactobacillus acidophilus , Streptococcus thermophilus , and Bifidobacterium bifidum. [ 0033 ] The term " lactobacillus "

In order to obtain a culture medium for lactic acid bacteria or a fermented product, lactic acid bacteria may be inoculated and cultured in a milk-containing raw material. The specific conditions of each step may be determined by various factors such as the type of lactic acid bacteria used and the desired fermented milk products However, it is not particularly limited.

In an embodiment of the present invention, there is provided a fermented dairy product comprising a composition for enhancing storage stability of the above cultured lactic acid bacteria, and a culture of a lactic acid bacterium produced by culturing a lactic acid bacterium in a milk-containing raw material. Fermented dairy products include, but are not limited to, fermented milk, yogurt, fresh cheese products, cheese including mozzarella cheese, and dairy products such as buttermilk.

The composition for enhancing storage stability according to the present invention has an advantage in that it can lower the content of sugars and fructose used in the conventional lactic acid bacteria culture solution and increase the storage stability of the lactic acid bacteria culture. In addition, it may not contain the pectin conventionally used to prevent the phase separation (phase separation) of the cultured lactic acid bacteria, and solves the problem due to the low solubility of the pectin. Even if the pectin is used in an amount smaller than the conventional amount, .

In one example, the fermented dairy product can further improve texture using modified starch. Modified starches usable in the present invention include starches such as oxidized starch, acetylated distarch adipate, acetylated distarch phosphate, starch sodium octenyl succinate, Phosphated Distarch Phosphate, Phosphated Distarch Phosphate, Starch Acetate, Hydroxypropyl Distarch Phosphate and Hydroxypropyl Starch Phosphate Starch Acetate, Hydroxypropyl Distarch Phosphate, ), And the like.

In accordance with a specific embodiment of the present invention, a process for producing a culture liquid of lactic acid bacteria containing a cyclosporin is described.

First, a process for producing a mixed solution is prepared by mixing a milk-containing raw material, a saccharide containing a saccharide or a rare saccharide, a modified starch, and the like.

 2 to 97 parts by weight of a milk-containing raw material, 2 to 20 parts by weight of a milk-containing raw material and 0.1 to 2 parts by weight of a modified starch, based on 100 parts by weight of the solid content of the whole mixed solution. The milk-containing raw material may include at least one selected from the group consisting of raw milk, low-fat milk, non-fat milk, reduced milk, reduced low fat milk and skim milk powder and, if cultured lactobacilli are cultivated appropriately, I never do that.

The mixed raw material is sterilized and cooled to obtain a mixed liquid at 38 to 43 ° C. This is for fermenting the fermented lactic acid bacteria smoothly, and when the sterilization and cooling liquid is low, the fermented lactic acid bacteria do not ferment smoothly, The fermentation time can be shortened when the cooling temperature of the sterilization and cooling liquid is more than 40 ° C, but the flavor of the culture product of the lactic acid bacteria, which is the final product of the present experiment, is decreased or the texture is changed.

After inoculating the lactic acid bacteria in the process of inoculating the sterilization and cooling solution with the lactic acid bacteria, the lactic acid bacteria are inoculated and cultured in the fermenter until the pH is reduced to 4.0 ~ 4.6 in the fermenter at 37 ~ 40 ℃. When the pH of the culture solution is less than pH 4.0, the viscosity and lactic acid of the culture solution of lactic acid bacteria are not constant and the texture and flavor are decreased due to excessive fermentation. When the pH value is more than 4.6, the lactic acid bacteria are not fermented enough, The texture is deteriorated.

If the pH of the culture solution is less than 4.0, the viscosity and lactic acid of the culture solution of lactic acid bacteria are not constant and the texture and flavor are decreased due to past fermentation. If the pH value is more than 4.6, the lactic acid bacteria can not sufficiently ferment, And the texture is deteriorated.

Wherein the cooling step is a step of cooling the culture liquid at 15 to 20 ° C.

After the culture solution is cooled, a homogenized solution is prepared by applying a pressure of 150 to 300 kg / cm ² (BAR) in a homogenizer. The homogenized solution is homogenized at a pressure of 150 to 300 kg / cm ² (BAR) in order to prevent the formation of a cream layer due to separation of milk fat in the culture product of lactic acid bacteria, which is the final product of the present experiment, If the milk fat is separated from the fermented milk, the cream layer is formed and the flavor is lowered. If the homogeneous pressure is too high, the whey protein of the lactic acid bacterium culture liquid, which is the final product of the present experiment, is separated and the flavor is lowered.

The fermented milk product is prepared by filling the fermented broth of lactic acid bacteria, which is the final product of the present experiment through homogenizer, in a refrigerator. Methods of producing conventional fermented dairy products, dilution and addition of various additional ingredients, and the like.

The composition for enhancing storage stability of a culture of a lactic acid bacterium containing a cycloserone according to the present invention has an advantage that the physical properties, taste and properties of a lactic acid bacterium cultured product or a fermented dairy product can be maintained within a certain range during a storage period. The lactic acid bacterium culture product or the fermented milk product according to the present invention may further comprise an additive selected from the group consisting of modified starch and pectin. The addition of the modified starch also has the effect of imparting a soft texture of the fermented milk. The pectin used in the lactic acid bacteria fermented product or the fermented milk product for the purpose of preventing the layer separation or the deterioration phase has a disadvantage that the solubility thereof is poor and the composition for enhancing the storage stability with the psicose according to the present invention, alone or in combination with pectin, Can be further improved.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a process diagram showing a method for producing a culture liquid of lactic acid bacteria containing D-psicose according to an embodiment of the present invention. FIG.
Fig. 2 is a diagram showing an example of an expression recombinant vector (pCES_sodCDPE) for producing a psicose syrup used in the present invention.
FIG. 3 is a graph showing changes in pH with time of culture according to an embodiment of the present invention. FIG.
4 is a graph showing changes in acidity with time of culture according to an embodiment of the present invention.
5 is a graph showing changes in pH of the culture solution of lactic acid bacteria according to the storage period according to an embodiment of the present invention.
6 is a graph showing changes in acidity of the culture solution of lactic acid bacteria according to the storage period according to an embodiment of the present invention.
FIG. 7 is a photograph showing Lee Soo-Hyun image after 5 days of storage of the culture solution of lactic acid bacteria according to an embodiment of the present invention.
FIG. 8 is a photograph showing Lee Soo Hyun image 10 days after storage of the culture solution of lactic acid bacteria according to an embodiment of the present invention.
FIG. 9 is a photograph showing Lee Soo Hyun's image after 15 days of storage of the culture solution of lactic acid bacteria according to an embodiment of the present invention.
FIG. 10 is a graph showing the percentage of completion (%) of the culture solution of lactic acid bacteria during the storage period according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Manufacturing example  One: Saikos  Production strains and Saikos  Manufacture of syrups

Manufacturing example  1-1: Saikos  Production of production strains

The DPE gene (Gene bank: EDS06411.1) derived from Clostridium scindens ATCC 35704 was synthesized as modified polynucleotides by optimizing E. coli for the Escherichia coli CDPE. The polynucleotide optimized for Escherichia coli (SEQ ID NO: 2) and the sod promoter and T7 terminator obtained from the pET21a vector were obtained through PCR using each template, which was then ligated into a template by overlap PCR (PCR) And cloned into pGEM T-easy vector through T-vector cloning to confirm the sequence of the polynucleotide including the sod promoter (SEQ ID NO: 1), the optimized CDPE sequence of SEQ ID NO: 8 and the T7-terminator.

The entire confirmed polynucleotide was inserted into the same restriction enzyme site of the expression vector pCES208 (J. Microbiol. Biotechnol., 18: 639-647, 2008) using restriction enzymes NotI and XbaI (NEB) to obtain a recombinant vector pCES208 / (PCES_sodCDPE) was prepared. The cleavage map of the prepared recombinant vector (pCES_sodCDPE) is shown in Fig.

The prepared recombinant vector (pCES_sodCDPE) plasmid was transformed into Corynebacterium glutaricum using electroporation. Colonies were picked and inoculated into 4 ml of LB medium (10 g / L of tryptone, 10 g / L of NaCl, 5 g / L of yeast extract) supplemented with kanamycin at a final concentration of 15 ug / And cultured for about 16 hours. Then, 1 ml of the culture was obtained and inoculated into 100 ml of LB medium containing 15 ug / ml kanamycin, and the cultivation was continued for 16 hours or more. After lysis of cells cultured with Beadbeater, only supernatant is obtained, mixed with sample buffer 1: 1, and heated at 100 ° C for 5 minutes. The prepared sample was subjected to electrophoresis on a 12% SDS-PAGE gel (composition: running gel - 3.3 ml H2O, 4.0 ml 30% acrylamide, 2.5 ml 1.5 M Tris buffer (pH 8.8) 20 μl 10% SDS, 20 μl 10% APS, 2 μl TEMED) at 180 V for about 50 minutes at room temperature The protein expression was confirmed by electrophoresis. After the expression of CDPE was confirmed on SDS-PAGE gel, the His-tag purification was performed using Ni-NTA resin to determine the precise expression level. The expression (%) = (Purified protein (mg) / Total soluble protein )) * 100). The transformed Corynebacterium glutaricum prepared above produced 16.62 mg of total soluble protein and 1.74 mg of purified enzyme protein.

Manufacturing example  1-2: Saikos  Produce

Cells were recovered by centrifugation in a strain culture in order to produce a scissors from fructose using the recombinant strain producing the scuche epimerase obtained in Production Example 1-1.

The cell suspension was then treated with 0.05% (w / v) of an emulsifier (M-1695) in a final volume at 35 ° C (± 5 ° C) for 60 minutes. After the completion of the reaction, the supernatant containing the emulsifier was removed using a centrifuge, and the cells were recovered.

For the preparation of the immobilized beads, the recovered cells were mixed with DW to a final cell concentration of 5% (w / v), mixed with 4% (w / v) alginic acid dissolved in water and 5% ) Were mixed at a ratio of 1: 1 and stored at 4 ° C in order to remove the bubbles generated during mixing. The mixture solution was injected through a Neddle (inner diameter: 0.20 to 0.30 mm) and formed into a droplet shape. The mixture was dropped by weight, and the dropped mixed solution was dropped into a previously prepared 100 mM calcium chloride (CaCl 2) Spherical or elliptical beads (diameter 2.0 to 2.2 mm) were formed. The formed beads were soaked in a 100 mM calcium chloride solution and mixed evenly by a stirrer so as to be further cured.

After all of the mixed solution was injected, the beads were further cured while refrigerated for 4 to 6 hours, and then replaced with a fresh 100 mM calcium chloride solution, and cured in a refrigerated state for about 6 hours. The hardened beads were completely removed from the beads, and then water having a volume three times the bead volume was added thereto. The beads were then stirred for 10 minutes. This process was repeated three times to remove the calcium chloride solution. The washed beads were completely removed from the water for manganese soaking, and then a 40-brix (%) reaction substrate containing 10 mM manganese was added in a volume three times as large as the bead volume and stirred for 10 minutes. And replaced with a reaction substrate containing manganese. The reaction substrate is adjusted to pH 6.8 ~ 7.2 with 3N NaOH. Depending on the type of product, liquid fructose or crystalline fructose can be the reaction substrate. After the beads were replaced with the reaction substrate containing 10 mM manganese, the beads were transferred to the reaction tank and reacted at a reaction temperature of 50 ° C. for about 30 to 60 minutes to complete the sorbing of beads with manganese and fructose. The diameter of the finished bead decreases to 1.6 ~ 1.8mm and the strength also increases. The substrate of the beads which had been soaked was removed and the immobilized reaction column was filled and used for the production of the viscose syrup.

≪ Immobilization column reaction conditions >

Reaction temperature: Internal temperature of column jacket 50 ℃

Substrate flow rate: 0.5 SV (space velocity L. h-1)

Reaction Substrate: Crystalline fructose 40brix, pH 6.8 ~ 7.2, Mn ^{2 +} free

Bead preparation: 2.5% (w / w) cells, 2% (w / w) alginic acid mixture and 10 mM Mn ^{2 +} soaking

The immobilization reaction column was provided with a raw material containing 92% by weight of fructose when the raw material solution contained 75% of solid content and the total solid content was 100% by weight, to prepare a psycho syrup. 25 (w / w)% of a sicos syrup was obtained from the reaction solution in a weight ratio of glucose: fructose: cyoses: oligosaccharides of 6: 67: 25: 2.

Example  1 to 2: Preparation of Lactic Acid Bacterium Culture Solution

A raw material mixture as shown in Table 1 below was prepared by mixing saccharide or psicose syrup with crude oil, skim milk powder and modified starch. In Table 1, glucose, psicose syrup and modified starch are products of Samyang Genex Co., Ltd., white sugar is a product of Samyang Co., and compound lactobacillus (ABT-5) is a product of Chr-Hansen. The above-mentioned psicose syrup was the 25% psicose syrup prepared in Preparation Example 1.

Ingredients Comparative Example 1 Comparative Example 2 Example 1 Example 2 Example 3 Crude oil (wt%) 65 65 65 65 65 Skim milk powder (wt%) 3.5 3.5 3.5 3.5 3.5 Glucose (wt%) 5 - - - - White sugar (wt%) - 5 - - - Psycosis syrup
25% - - 5 6.8 7.0 Pectin (wt%) 0.05 0.05 0.05 - - Modified starch (wt%)
(Neo HP) 0.15 0.15 0.15 0.2 - Complex lactic acid bacteria
(ABT-5) (wt%) 0.005 0.005 0.005 0.005 0.005 Purified water (wt%) Balance Balance Balance Balance Balance Sum (wt%) 100 100 100 100 100

The obtained raw material mixture solution was sterilized at 90 to 95 캜 for 5 to 10 minutes, and the sterilized raw material solution was cooled to 38 to 43 캜.

To each of the above-mentioned sterilized and cooled mixed liquor, Lactobacillus ( Lactobacillus) acidophilus), were inoculated with ABT-5 of Streptococcus Thermo filler's (Streptococcus thermophilus) and Bifidobacterium bipyridinium bushes (Chr.-Hansen Co. (Denmark) is mixed Bifidobacterium bifidum). After the lactic acid bacteria were inoculated, the cells were cultured in a fermenter at 37 to 40 DEG C until the pH was reduced to 4.0 to 4.6, and then the culture was cooled to 15 to 20 DEG C. The cooled culture was subjected to a pressure of 200 Kg / cm ² (BAR) using a homogenizer (NIRO SOAVI, Italy) to prepare a homogenate. A culture solution of lactic acid bacteria supplemented with psicose syrup was prepared.

Comparative Example  1 and 2: Production of Lactic Acid Bacterium Culture Solution

A raw material liquid was prepared in substantially the same manner as in Example 1 except that 5 weight% of grape was used in Comparative Example 1 and 5 weight% of white sugar was used in Comparative Example 2 instead of the psicose syrup of Example 1. [ The raw material compositions of Comparative Examples 1 and 2 are shown in Table 1 above.

The same lactic acid bacterium ABT-5 as in Example 1 was inoculated to each of the above-mentioned sterilized and cooled mixed liquor, followed by incubation, cooling, and homogenization to prepare a lactic acid bacterium culture solution prepared by adding glucose or white sugar instead of the cyclosporin.

Test Example  1: acidity of cultured lactic acid bacteria and pH  Measure

The pH and acidity of the culture broth of lactic acid bacteria obtained in Examples 1, 2 and 3 and Comparative Examples 1 and 2 were measured.

Specifically, the pH of fermented lactic acid bacteria was measured before fermentation, 6 hours after fermentation, and after homogenization using a pH meter (Methrom 780 pH meter, Switzerland).

The acidity was measured by taking 10 ml of each sample and 10 ml of distilled water from which CO ₂ had been removed. The pH of the solution was adjusted to 8.5 with 0.1 N NaOH. The acidity of lactic acid was calculated by the following equation.

[Calculation of acidity]

The results measured by the above formula are shown in Table 3 below.

The pH and acidity before and after the homogenization were measured to confirm the quality of the cultured lactic acid bacteria and to confirm changes in the culture and homogenization process before and after the lactic acid bacteria culturing and the homogeneity of pH and acidity .

Quality measurement Comparative Example 1 Example 1 Comparative Example 2 Example 2 Example 3 Pre-incubation pH 6.411 6.419 6.384 6.430 6.387 After completion of incubation (6 hours) pH 4.448 4.463 4.455 4.446 4.451 PH after homogenization 4.290 4.303 4.332 4.296 4.298 Pre-culture pH (%) 0.125 0.125 0.125 0.125 0.125 After the incubation (6 hours), the acidity (%) 0.752 0.752 0.752 0.755 0.752 Acidity after Homogenization (%) 0.743 0.740 0.743 0.752 0.740

As a result of the measurement of the pH and acidity, it was confirmed that the measurement items after the production of the lactic acid bacterium culture solution were all similar, and that the pH and acidity of the lactic acid bacterium culture solution according to the present invention remained at the same level as the conventional product during the storage period after the culture.

Test Example  2: acidity and pH  Measure

The pH and acidity of the culture broth of lactic acid bacteria obtained in Examples 1, 2 and 3 and Comparative Examples 1 and 2 were measured. The end point of the cultivation of the lactic acid bacteria is usually determined by pH and acidity. For example, the pH is in the range of 4.4 to 4.6 and the acidity is in the range of 0.7 to 0.8%, and may be slightly different depending on the producer and the product.

PH changes and acidity changes of the culture broth with the lapse of culture time were measured in the production process of the culture broth of lactic acid bacteria obtained in Examples 1, 2 and 2 and Comparative Examples 1 and 2.

The pH of the fermentation broth of the lactic acid bacteria was measured in the same manner as in Test Example 1, and the change with time was measured every 1 hour from 0, 1, 2, 3, 4, 5, 6, 7 and 8 hours from the start of fermentation Respectively. The measurement results are shown in Table 4 and FIG. 3 below.

division Comparative Example 1 Example 1 Comparative Example 2 Example 2 Example 3 0 hours 6.411 6.419 6.384 6.430 6.387 1 hours 6.195 6.210 6.124 6.267 6.204 2 hours 5.504 5.448 5.232 5.531 5.483 3 hours 4.800 4.833 4.727 4.820 4.818 4 hours 4.534 4.562 4.550 4.536 4.551 5 hours 4.480 4.490 4.485 4.497 4.497 6 hours 4.448 4.463 4.455 4.446 4.451 7 hours 4.253 4.272 4.316 4.372 4.348 8 hours 4.182 4.199 4.250 4.217 4.225

As shown in Table 4, it was confirmed that the pH change was similar in glucose (Comparative Example 1), Sugar (Comparative Example 2) and PsycOS syrup (Example) depending on the culture time. Table 3 shows the degree of cultivation of lactic acid bacteria when added with sicos containing sicos compared to glucose and white sugar. And the pH change was measured by the incubation time. As a result, it was found that the result did not decrease the production efficiency during cultivation.

The acidity of the fermented lactic acid bacteria fermented was measured in the same manner as in Test Example 1, and the change over time was measured every 1 hour from 0, 1, 2, 3, 4, 5, 6, 7 and 8 hours from the start of fermentation Respectively. The measurement results are shown in Table 5 and Fig.

division Comparative Example 1 Example 1 Comparative Example 2 Example 2 Example 3 0 hours 0.125 0.125 0.125 0.125 0.125 1 hours 0.180 0.180 0.198 0.180 0.180 2 hours 0.360 0.330 0.450 0.360 0.360 3 hours 0.585 0.585 0.608 0.585 0.585 4 hours 0.675 0.653 0.680 0.672 0.653 5 hours 0.710 0.698 0.719 0.705 0.705 6 hours 0.716 0.723 0.743 0.743 0.723 7 hours 0.765 0.765 0.799 0.789 0.765 8 hours 0.855 0.833 0.825 0.810 0.825

As shown in Table 5, it was confirmed that the change of acidity was high in the order of glucose (Comparative Example 1), sugar (Comparative Example 2) and 25% (Example).

Test Example  3: Sensory evaluation of culture

The flavor components produced by lactic acid bacteria are diactyl, acetoin, volatile acids, carbon dioxide, acetaldehyde, ethanol and so on. Therefore, the degree of the occurrence of these flavor components after the culture was confirmed by sensory evaluation.

The number of sensory evaluation panels was 12, and the sensory evaluation method was given from 10 points to 7 items by 10 point scale method.

[Evaluation Items]

The degree of sharpness of the taste (not very sharp (1 point) - very sharp (10 points)),

The softness of the taste (not very soft (1 point) - very soft (10 points)),

(1 point) - No fermentation at all (10 points)), lactic acid fermentation (diacetyl alcohol, aldehyde alcohol, etc.)

The degree of acidity (no acidity (1 point) - very strong acidity (10 points)),

Degree of sweetness (there is no sweetness at all (1 point) - sweetness is very strong (10 points)),

(Not at all tangible (1 point) - very tough (10 points)),

Overall likelihood (very bad (1 point) - very good (10 points))

The results of the sensory evaluation are shown in Table 6 below.

Evaluation items Comparative Example 1 Example 1 Comparative Example 2 Example 2 Example 3 Sharpness 7.4 5.9 5.5 3.8 3.2 Softness 4.0 4.6 5.2 6.0 6.7 Fermented lactic acid 5.8 5.1 5.4 4.6 4.5 Degree of acidity 5.1 4.5 4.4 3.7 4.2 Degree of sweetness 2.8 3.4 5.2 5.3 2.7 Aubiousness of the tail 6.6 5.3 4.7 4.2 4.0 Overall likelihood 5.5 6.2 5.4 6.9 6.8

As shown in Table 6, the degree of sharpness, softness, and sourness of glucose (Comparative Example 1) and Sugar (Comparative Example 2) was high and that of Cypress Syrup (Examples 1 and 2) was improved Could. The degree of sweetness was similar to that of sugar (Comparative Example 2) and 25% (Example 2) of psicose syrup, and the overall acceptability was improved when psicose syrup was added (Examples 1 and 2).

Test Example  4: Storage period of culture liquid of lactic acid bacteria pH  And acidity change measurement

The pH change and acidity change of the culture broth of lactic acid bacteria obtained in Examples 1, 2 and 3 and Comparative Examples 1 and 2 according to the storage period were measured.

For the cultured lactic acid bacteria obtained in Examples 1, 2, and 3 and Comparative Examples 1 and 2, the storage period was stored for 20 days in a refrigerator, and the storage was started on day 1, day 2, day 5, day 15, The pH of the culture medium was measured during the storage period using a 5-times pH meter (Methrom 780 pH meter, Switzerland), and the results are shown in Table 7 and FIG.

storage duration Comparative Example 1 Example 1 Comparative Example 2 Example 2 Example 3 1 day 4.290 4.303 4.332 4.296 4.294 2 days 4.285 4.303 4.329 4.320 4.302 5 days 4.187 4.200 4.243 4.248 4.277 10 days 4.140 4.167 4.215 4.213 4.216 15th 4.109 4.115 4.177 4.189 4.165

As shown in Table 7, it was confirmed that the pH changes during storage were all similar. For the cultured lactic acid bacteria obtained in Examples 1, 2, and 3 and Comparative Examples 1 and 2, the storage period was stored for 20 days in a refrigerator, and the storage was started on day 1, day 2, day 5, day 15, 5 times, add 90 ml of distilled water to 10 ml of sample, dilute 10-fold, add 2 ~ 3 drops of 1% phenolphthalein, titrate with 1 / 10N NaOH until the pH becomes 8.5, Respectively. Shown in Table 8 and FIG. 6 below.

[Calculation of acidity]

storage duration Comparative Example 1 Example 1 Comparative Example 2 Example 2 Example 3 1 day 0.743 0.740 0.743 0.752 0.743 2 days 0.807 0.784 0.782 0.765 0.782 5 days 0.855 0.824 0.844 0.810 0.800 10 days 0.878 0.833 0.855 0.833 0.825 15th 0.923 0.889 0.900 0.855 0.855

As shown in Table 8, it was confirmed that the acidity change in the storage period was the largest in glucose (Comparative Example 1), and the acidity change was small in the order of white sugar (Comparative Example 2) and 25% . It was confirmed that the rate of increase of acidity of Comparative Example 1 (glucose) and Comparative Example 2 (sugar) in the addition of D-psicose syrup was relaxed and inhibited increase of acidity during the storage period, And confirmed the fire resistance.

From the above results, it was confirmed that the increase of the acidity of the culture solution of lactic acid bacteria during the storage period was inhibited, and as a result, it was confirmed that the culture medium of lactic acid bacteria decreased the quality deterioration during the storage period Respectively.

Test Example  5: culture solution During storage  Lee Soo Hyun phase measurement

5-1: Evaluation of the qualities of this liquid

Day, 2 days, 5 days, 15 days, 20 days, and 20 days, respectively, based on the storage start time, while maintaining the fermented broth of lactic acid bacteria obtained in Examples 1 and 2 and Comparative Examples 1 and 2 in a refrigerator At the first day, the presence or absence of the occurrence of the Lactobacillus cultures was visually confirmed and photographed five times. The results are shown in Table 9 and Figs. 7 to 9. This is a phenomenon caused by the change of isoelectric point in the culture medium. As a result of visual observation, whey was separated into three stages, and three points, two points and one point Respectively. That is, 3 points are preferable results because there is almost no separation and isoform phase, and 1 point is a bad result when the sulfur and separation phenomenon is severe during the storage period.

storage duration Comparative Example 1 Example 1 Comparative Example 2 Example 2 Example 3 1 day 3 3 3 3 3 2 days 3 3 3 3 3 5 days One 3 2 3 3 10 days One 2 One 2 2 15th One 2 One 2 2

As can be seen from the above Table 9, the biodegradation of the culture broth occurred at the bottom of storage at the 5th day when glucose was applied (Comparative Example 1), and the biodegradation was also very severe. When the sugar was applied (Comparative Example 2), water was collected on the 10th day of storage, and it was confirmed that the water phase occurred especially in the upper part. It was confirmed that only slight dehydrated phase occurred in the upper layer on the 10th day of storage at the time of application of the psicose syrup (Example 1 and Example 2).

As a result, it was confirmed that when the glucose or sucrose was applied to the case of the sicos syrup, the resulting syrupy phase could be remarkably improved.

5-2: Quantitative evaluation of this liquid

For the culture of the lactic acid bacteria obtained in Examples 1, 2 and 3 and Comparative Examples 1 and 2, the culture of the lactic acid bacteria of each sample was accurately placed on a graduated scale in a 100 mL measuring cylinder, The degree of the generation of the liquid was measured at intervals of one day, that is, when the culture broth of the lactic acid bacteria was stored at 4 to 8 ° C, 1 day, 5 days, 10 days, and 15 days after storage. During the storage period, this liquid was calculated according to the following formula, and the amount of this liquid during the storage period was measured and shown in Table 10.

[Lee, Soo Hyun, Mathematical Expression]

 (%) Of the culture liquid = volume of the liquid generated / total volume of culture liquid of lactic acid bacteria × 100

In Table 10, the experimental result is v / v%.

storage duration Comparative Example 1 Example 1 Comparative Example 2 Example 2 Example 3 1 day 0 0 0 0 0 2 days 2 0 0 0 0 5 days 5 0 2 0 0 10 days 10 2 6 One One 15th 10 2 10 2 2

It was confirmed that the effect of adding D-psicose syrup was remarkably inhibited when compared with Comparative Example 1 (glucose) and Comparative Example 2 (sugar). 5 days after the application of the glucose of Comparative Example 1, water was collected at the lower end, and the water content was also very severe, and 10 days after the application of the sugar of Comparative Example 2, water was generated. In the case of the application of the psicose syrup, a slight dehydration occurred in the upper layer after 10 days. As a result, it was possible to improve Lee Soo Hyun 's image when applied with.

Usually, the lipid phase of lactic acid bacteria culture is a phenomenon which occurs when the pH is lowered and the isoelectric point (the pH of the milk isoelectric point is about 4.6) is changed by the charge of the cation or the anion, which is a typical phenomenon which causes deterioration of the quality of the culture liquid.

As a result of the above-mentioned various experimental examples, it was found that the addition of D-psicose syrup had an effect on the lipophilicity in comparison with that of Comparative Example 1 (glucose) and Comparative Example 2 (sugar) It could be a factor of the quality index and it was found that the rate of increase of the acidity was relaxed as compared with Comparative Example 1 (glucose) and Comparative Example 2 (sugar) when D-psicose was added.

<110> SAMYANG GENEX CORPORATION <120> composition for increasing storage stability of culture          product of lactic acid bacteria <130> DPP20145683KR <160> 9 <170> Kopatentin 1.71 <210> 1 <211> 356 <212> DNA <213> Artificial Sequence <220> The sod promoter (6) <400> 1 aagcgcctca tcagcggtaa ccatcacggg ttcgggtgcg aaaaaccatg ccataacagg 60 aatgttcctt tcgaaaattg aggaagcctt atgcccttca accctactta gctgccaatt 120 attccgggct tgtgacccgc tacccgataa ataggtcggc tgaaaaattt cgttgcaata 180 tcaacaaaaa ggcctatcat tgggaggtgt cgcaccaagt acttttgcga agcgccatct 240 gacggatttt caaaagatgt atatgctcgg tgcggaaacc tacgaaagga ttttttaccc 300 atggctgtat acgaactccc agaactcgac tacgcatacg acgaaaggat tacaaa 356 <210> 2 <211> 93 <212> DNA <213> Artificial Sequence <220> <223> The Tac1 promoter (4) <400> 2 tgacaattaa tcatcggctc gtatattgtg tggaattgtg agcggataac aatttcacac 60 aggaaacaga attcccgggg aaaggattac aaa 93 <210> 3 <211> 112 <212> DNA <213> Artificial Sequence <220> <223> The Tac2 promoter (4) <400> 3 tgacaattaa tcatccggct cgtataatgt taacaatttg tggaattgtg agcggacaca 60 caggaaacag accatggaat tcgagctcgg tacccgggga aaggattaca aa 112 <210> 4 <211> 30 <212> DNA <213> Artificial Sequence <220> The Trc promoter (1) <400> 4 tgacaattaa tcatcggcct cgtataatgt 30 <210> 5 <211> 7 <212> DNA <213> Artificial Sequence <220> <223> Ribosome binding region <400> 5 gaaagga 7 <210> 6 <211> 7 <212> DNA <213> Artificial Sequence <220> <223> Spacer sequence <400> 6 ttacaaa 7 <210> 7 <211> 289 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of an enzyme protein originated from          Clostridium scindens <400> 7 Met Lys His Gly Ile Tyr Tyr Ala Tyr Trp Glu Gln Glu Trp Ala Ala   1 5 10 15 Asp Tyr Lys Arg Tyr Val Glu Lys Ala Ala Lys Leu Gly Phe Asp Ile              20 25 30 Leu Glu Val Gly Ala Ala Pro Leu Pro Asp Tyr Ser Ala Gln Glu Val          35 40 45 Lys Glu Leu Lys Lys Cys Ala Asp Asp Asn Gly Ile Gln Leu Thr Ala      50 55 60 Gly Tyr Gly Pro Ala Phe Asn His Asn Met Gly Ser Ser Asp Pro Lys  65 70 75 80 Ile Arg Glu Glu Ala Leu Gln Trp Tyr Lys Arg Leu Phe Glu Val Met                  85 90 95 Ala Gly Leu Asp Ile His Leu Ile Gly Gly Ala Leu Tyr Ser Tyr Trp             100 105 110 Pro Val Asp Phe Ala Thr Ala Asn Lys Glu Glu Asp Trp Lys His Ser         115 120 125 Val Glu Gly Met Gln Ile Leu Ala Pro Ile Ala Ser Gln Tyr Gly Ile     130 135 140 Asn Leu Gly Met Glu Val Leu Asn Arg Phe Glu Ser His Ile Leu Asn 145 150 155 160 Thr Ser Glu Glu Gly Val Lys Phe Val Thr Glu Val Gly Met Asp Asn                 165 170 175 Val Lys Val Met Leu Asp Thr Phe His Met Asn Ile Glu Glu Ser Ser             180 185 190 Ile Gly Asp Ala Ile Arg His Ala Gly Lys Leu Leu Gly His Phe His         195 200 205 Thr Gly Glu Cys Asn Arg Met Val Pro Gly Lys Gly Arg Thr Pro Trp     210 215 220 Arg Glu Ile Gly Asp Ala Leu Arg Glu Ile Glu Tyr Asp Gly Thr Val 225 230 235 240 Val Met Glu Pro Phe Val Arg Met Gly Gly Gln Val Gly Ser Asp Ile                 245 250 255 Lys Val Trp Arg Asp Ile Ser Lys Gly Ala Gly Glu Asp Arg Leu Asp             260 265 270 Glu Asp Ala Arg Arg Ala Val Glu Phe Gln Arg Tyr Met Leu Glu Trp         275 280 285 Lys     <210> 8 <211> 870 <212> DNA <213> Artificial Sequence <220> Modified nucleic acid sequence (1) of the enzyme protein of SEQ          ID NO: 7 <400> 8 atgaaacacg gtatctacta cgcgtactgg gaacaggaat gggcggcgga ctacaaacgt 60 tacgttgaaa aagcggcgaa actgggtttc gacatcctgg aagttggtgc ggcgccgctg 120 ccggactact ctgcgcagga agttaaagaa ctgaaaaaat gcgcggacga caacggtatc 180 cagctgaccg cgggttacgg tccggcgttc aaccacaaca tgggttcttc tgacccgaaa 240 atccgtgaag aagcgctgca gtggtacaaa cgtctgttcg aagttatggc gggtctggac 300 atccacctga tcggtggtgc gctgtactct tactggccgg ttgacttcgc gaccgcgaac 360 aaagaagaag actggaaaca ctctgttgaa ggtatgcaga tcctggcgcc gatcgcgtct 420 cagtacggta tcaacctggg tatggaagtt ctgaaccgtt tcgaatctca catcctgaac 480 acctctgaag aaggtgttaa attcgttacc gaagttggta tggacaacgt taaagttatg 540 ctggacacct tccacatgaa catcgaagaa tcttctatcg gtgacgcgat ccgtcacgcg 600 ggtaaactgc tgggtcactt 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Claims (21)

A composition for enhancing storage stability of a culture of a lactic acid bacterium, The composition of claim 1, wherein the storage stability of the lactic acid bacterium culture is maintained during pH and acidity conditions during storage and the phase is prevented. The composition according to claim 1, wherein the lactic acid bacterium culture product has a completion rate of 5 v / v% or less expressed as a percentage of the volume of the liquid generated per volume of the whole lactic acid bacteria culture culture during the storage period. The composition according to claim 1, wherein the lactic acid bacterium culture product has a completion rate of 5 v / v% or less expressed as a percentage of the volume of the liquid generated per volume of the whole lactic acid bacterium culture culture during the storage period. The composition of claim 1, wherein the psicose is in powder or liquid form. 2. The composition of claim 1 comprising from about 2% to about 55% by weight, based on 100% by weight of the total solids content of the composition, of Saikos. The composition according to claim 1, wherein the Saicos is a mixed sugar comprising 2 to 55 parts by weight of Psicos, 30 to 80 parts by weight of fructose, and 2 to 60 parts by weight of glucose based on 100 parts by weight of the solid content. The composition of claim 1, wherein said lactic acid bacterium culture is a culture produced by culturing a lactic acid bacterium in a milk-containing raw material. Using Saikos,
A method for increasing the storage stability of cultured lactic acid bacteria using lactic acid bacteria and milk-containing raw materials. [10] The method of claim 9, wherein the lactic acid bacterium culture product has a completion rate of 5 v / v% or less expressed as a percentage of the volume of the liquid generated per volume of the whole lactic acid bacteria culture during the storage period. 10. The method according to claim 9, wherein the cyclosporine is added to the raw material composition for the production of the lactic acid bacteria culture, or is added to the lactic acid bacteria culture. 10. The method of claim 9, wherein the psicose is in powder or liquid phase. 10. The method of claim 9, wherein the psicose comprises from about 2% to about 55% by weight of psicose based on 100% by weight of the total composition. 10. The method according to claim 9, wherein the cyclosporine is mixed with 2 to 55 parts by weight of sucrose, 30 to 80 parts by weight of fructose and 2 to 60 parts by weight of glucose based on 100 parts by weight of the total solid content of the mixed sugar. 10. The method according to claim 9, wherein the milk-containing raw material comprises at least one selected from the group consisting of raw milk, low fat milk, non-fat milk, reduced milk, reduced low fat milk and skim milk powder. A fermented dairy product comprising a composition for enhancing storage stability of a culture of a lactic acid bacterium according to any one of claims 1 to 8 and a culture of a lactic acid bacterium produced by culturing a lactic acid bacterium in a milk-containing raw material. 17. The fermented milk product according to claim 16, wherein the fermented milk product does not contain pectin. The fermented milk product according to claim 16, wherein the dairy product further comprises at least one selected from the group consisting of modified starch and pectin. The fermented milk product according to claim 16, wherein the milk-containing raw material comprises at least one selected from the group consisting of raw milk, low fat milk, non-fat milk, reduced milk, reduced low fat milk and skim milk powder. 16. The fermented milk product according to claim 16, wherein the fermented milk product is prepared by culturing lactic acid bacteria in a raw material composition containing 2 to 97 parts by weight of a milk-containing raw material and 2 to 20 parts by weight of a sucrose based on 100 parts by weight of the total solid content of the fermented milk product Fermented dairy products. The fermented milk product according to claim 16, wherein the fermented milk product comprises 2 to 97 parts by weight of milk-containing raw material, 2 to 20 parts by weight of sucrose and 0.1 to 2 parts by weight of modified starch, based on 100 parts by weight of the total solid content of the fermented milk product A fermented dairy product produced by culturing a lactic acid bacterium in a raw material composition.

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