TR2023004151A2 - NANOENCAPSULATED YOGURT STARTER CULTURE PRODUCTION METHOD - Google Patents

Abstract

Meet; It is related to nanotechnology and yoghurt starter culture production method in yoghurt production in the dairy industry.

Description

DESCRIPTION: NANOENCAPSULATED YOGURT STARTER CULTURE PRODUCTION METHOD TECHNICAL FIELD: The invention relates to the nanotechnology-based yogurt starter culture production method in the dairy sector. KNOWN STATE OF THE TECHNOLOGY: Commercial yogurt produced in our country is fermented with imported culture. In rural areas, our people, having learned from their ancestors, recreate yogurt production by adding a portion of the previous yogurt to milk. However, with the recent migration from rural areas to urban centers, this tradition has begun to fade away. Consequently, almost all of our people are forced to continue consuming yogurt produced with imported starter cultures. The invention, titled "Production of Fermented Dairy Products from Natural Isolate Starter Cultures," published in the state of the art and numbered TR2020/03267, allows the isolation and purification of Streptococcus thermophilus and Lactobacillus bulgaricus, lactic acid bacteria naturally found in local village yogurts in Anatolia, and their commercialization in ready-to-use form. This process utilizes starter cultures containing different combinations of these bacteria to produce fermented dairy products such as creamy yogurt, homogenized yogurt, yogurt with various fat/dry matter contents, and ayran. This invention utilizes completely natural isolates isolated from yogurts collected from local villages in Anatolia, without any modifications. A sustainable method has been developed that is industrially applicable for mass production and meets the standard structure and taste desired by consumers in our country. The invention produces naturally fermented dairy products using starter cultures, commonly known as yeast/live yogurt cultures in Turkish Lira and the Turkish Food and Agriculture Organization (TGK) Nutrition and Health Declarations Regulation. The patent application states that the starter culture can be used in industrial mass production. No information is provided regarding its suitability for batch systems. The invention titled "Starter Culture Suitable for Yogurt Production Obtained from Sources" relates to pure strains of Streptecoccus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, which were obtained from traditional yogurts used in yogurt production and did not contain commercial cultures and were determined to have starter culture properties after technological characterization tests. In the 1st and 2nd patent applications, it is not stated whether any process was applied or not to the isolates obtained from local sources to create commercial cultures. In the state of the art, there is no publication or catalog that includes the differences such as the local yogurt starter culture used, the specific properties of the culture isolates, the type of nanotechnological process applied for the commercialization of the culture, the type and mixing ratios of the materials used in nanotechnology, the device conditions used in nanotechnology and the product trial of the created culture. In the state of the art, yogurt starter cultures are sold online. However, these cultures are not available in the local It is not. It consists of imported culture isolates produced by distributors of foreign companies in our country. On the other hand, there are also probiotic isolates (Lactobacillus acidophilus and Bifidobacterium sp.) added to the culture content to make it probiotic. For example, the Vivo brand is of Ukrainian origin. Recently: R&D studies have been carried out for local yogurt culture production in provinces such as Konya and Bursa. However, the products they produce generally use the frozen lyophilized method. In this method, the viability rate is low, and cold chain conditions must be ensured during transportation and storage. Due to the problems described above and the inadequacy of existing solutions on the subject, it has become necessary to make a development in the relevant technical field. DESCRIPTION OF THE INVENTION The present invention aims to eliminate the above-mentioned disadvantages and to bring new advantages to the relevant technical field, thus improving the yogurt industry in the dairy sector. Our goal in yogurt starter culture production, which has been developed in the production area and uses nanotechnology, is to create a commercial yogurt starter culture by nanoencapsulating isolates with the potential to be yogurt culture from Lactobacillus delbrueckii spp bulgaricus and Streptococcus thermophilus bacteria obtained from local sources using electrospinning. The starter culture obtained in our invention is suitable for use in both batch and batch systems. Our product is suitable for transportation and storage in different temperature environments. In our invention, a nanotechnological process has been applied that does not cause any change in the basic properties of the isolates obtained from local sources, but preserves their viability for a long time by protecting them from adverse environmental conditions such as transportation and storage. The product obtained in our invention can be stored under refrigerator conditions like other products, as well as having both transportation and storage properties at room temperature. This difference is an advantage of the nanotechnological process applied to culture. This is shown in Table 1. Drawings The applications of the present invention, briefly summarized above and discussed in more detail below, can be understood by referring to the example applications of the invention depicted in the attached drawings. However, it should be noted that the attached drawings only depict typical applications of the invention and are not to be considered limiting its scope, as the invention may therefore allow for other equally effective applications. Figure 1: SEM analysis image of the diameter size measurement of the nanoencapsulated material. Figure 2: TEM analysis image of the nanoencapsulated material at 50 nm. Figure 3: FT1R measurement graph of salep, WPl and nanomaterial (The top graph belongs to salep, the middle graph belongs to nanomaterial and the bottom graph belongs to WPl). Figure 4: This is the result of TGA analysis of nanoencapsulated material. To facilitate understanding, identical reference numbers have been used where possible to indicate identical elements common to the figures. The figures are not drawn to scale and may be simplified for clarity. It is believed that the elements and features of one application can be usefully incorporated into other applications without further explanation. DETAILED DESCRIPTION OF THE INVENTION In this detailed description, preferred alternatives to the nanoencapsulated yogurt starter culture production method of the invention are explained solely for the purpose of better understanding the subject and in a way that does not create any limiting effects. The yogurt starter culture produced as a result of the invention is suitable for use in the dairy industry. It can be used in both industrial and home yogurt production. The application is carried out by adding directly to pretreated milk at a ratio of 0.5-1.0 g culture/1 L milk. is made. It is also suitable for use by using an intermediate culture to shorten the incubation period on an industrial scale. Our invention is a yogurt starter culture product created with a new method. 1) The traditional yogurt starter culture bacteria used in our invention are Lactobacillus delbrueckeii subsp. bulgaricus and Streptococcus thermophilus. The bacteria in question were isolated from local yogurt products around Erzurum. 2) Whey powder was used in our invention. The relevant product is called 90% demineralized whey powder (90g protein/100g). 3) Salep powder was used in our invention. Studies have shown that the composition of salep powder generally consists of 7-61% glucomannan, 1-36% starch, 0.5-1% nitrogenous matter and 0.2-6% ash. 4) The electrospinning device is designed with a sensitivity of 0-30 kV and 0-1 mA. The device operates according to the desired specifications. ) The milk used in yogurt production was taken after being standardized for yogurt production. The pH value of the milk was 6.65, the fat content was 3.6%, and the dry matter content was 3.6%. As the first step in the invention, a 4% salep solution was prepared. The solution was stirred at room temperature at 500 rpm for 2 hours to ensure complete dissolution of the glucomannan in the salep. As the second step, centrifugation was carried out at 1500 rpm for 20 minutes to separate the dissolved glucomannan. After centrifugation, a pellet was added to obtain a solution containing 2% glucomannan. 40% whey protein isolate was added to the resulting glucomannan solution. (WPl) is added. WPl is denaturated by stirring at 600 rpm for 30 minutes at 85-90°C (Salep Glucomannani; 0.1-5.0%, WPl was mixed at 7.0-50.0%. The most suitable concentration ratios were 2.0% salep Glucomannani and 7.0-50.0%. As a result of these processes, nanoencapsulation material was formed. The ideal electrospinning device conditions for the formation of nanoencapsulation material were determined (11 kV high power application, 0.2 ml/h flow rate and 12 cm distance between the needle and the collector plate). The structure and function characterization of the nanoencapsulated material; SEM, TEM, FT1R and TGA analysis results were evaluated, and the formed material was In the nanoencapsulation of yogurt bacterial isolates, yogurt bacterial isolates (Lactobacillus delbrueckeii subsp. bulgaricus and Streptococcus thermophilus) with a bacterial count of 107-109 cfu/ml were added to the nanoencapsulation solution cooled to room temperature and mixed at 200 rpm for another 30 minutes to ensure homogeneous distribution of the bacteria. The nanoencapsulation process was carried out in the electrospinning device that provided the most ideal conditions. The viability stability and solubility efficiency of the nanoencapsulated culture in different environments (bile and gastric fluid, low pH, different temperature environments (-20, +4 and °C) were determined (Tables 1, 2, 3 and 4). Finally, yogurt was produced using a nanoencapsulated starter culture, and physicochemical, microbiological, and sensory analyses of the yogurt were performed. The results were compared with the literature (Tables 5-11). Our findings: o Nanoencapsulation was used for the first time in yogurt starter culture production, o Native yogurt bacteria were encapsulated at nanoscale for the first time as a starter culture, o Salep glucomannan was used for the first time in electrospinning, o Electrospinning conditions for WPL and salep glucomannan were optimized, o Nanoencapsulation significantly preserved bacterial viability under gastrointestinal conditions, o Nanoencapsulation did not delay the incubation time of the starter culture, o The nanoencapsulated starter culture increased acetaldehyde production in yogurt samples by fourfold, o The fact that nanoencapsulation preserved bacterial viability best at room temperature, and that the nanoencapsulated culture maintained approximately 1.5-2 log more bacterial viability compared to the control group after 28 days of storage, constitutes a novel and inventive step. Since the bacterial isolates in our invention were obtained from local products, this ensured that the final product was local (national). Whey protein isolate (WPl) has a high protein content, increasing the applicability of the method used. Furthermore, its high protein content improved the physicochemical properties of the product. Salep glucomannan formed a homogeneous polymer with WPl, assisting in the electrospinning of WPl. On the other hand, salep glucomannan, which is said to be a prebiotic, provided higher viability rates of the bacterial isolates. The nanoencapsulation The material significantly enriched the aroma profile of the final product, yogurt. The electrospinning device used in the invention enabled bacterial isolates to be encapsulated at nanoscale with the resulting solution. Finally, yogurt production was carried out with the produced nanoencapsulated starter culture, and physicochemical, microbiological, and sensory analyses of the yogurt were conducted. In our study, yogurt starter culture bacteria were nanoencapsulated with the electrospinning device. In developing the method, both the mixing ratios of the encapsulation materials and the application conditions of the electrospinning device must be optimized together. Changes in the mixing ratios of the materials or any parameter of the electrospinning device cause changes in the physicochemical and morphological properties of the final product. In the method we used in our study, yogurt starter culture bacteria were nanoencapsulated together at nanoscale for the first time, and salep glucomannan, known to have prebiotic properties, was also used for the first time. was used in the electrospinning method. The high viability of the nanoencapsulated culture created as a result of the studies performed; both the physicochemical and functional properties of the substances used in nanoencapsulation and the positive results of the product trials conducted with the culture indicate that the culture can be a local commercial yogurt starter culture. The component ratios in 100ml of milk used in yogurt production are; - Nanoencapsulated starter culture ratio is 0.5-2%, v/v (5ml-20ml), - % total dry matter of milk is 13.5% (g/ml), - % water ratio of milk is 85.98-86.10% (ml). Since nanoencapsulated starter culture varies between 0.025-0.1% by mass in 1 liter milk sample, it was ignored in the calculation of total dry matter. Tables and charts related to the subject of the invention are given below. Table 1. Viability stability of nanoencapsulated starters in different environments. Storage Abl AsZ Time Storage Temperatures A-B: Letters indicate that there is a statistically significant difference between the number of microorganisms in the sample at different storage temperatures in the same storage period (P<0.05). a-c: Letters indicate that there is a statistically significant difference between the number of microorganisms in the samples at different storage temperatures in the same storage periods (P<0.05). Table 2: Viability stability of nanoencapsulated starters in low pH environment. A-B: Letters indicate that there is a statistically significant difference (P<0.05) between the number of microorganisms in the samples at different pH environments in the same storage periods (P<0.05). Table 3: Viability stability of nanoencapsulated starters in bile salt environment. A-B: Letters indicate that there is a statistically significant difference in the same sample in different concentrations of bile salts at the same time. Table 4: Viability stability of nanoencapsulated starters in simulated gastric fluid. Time 03% Pepsin 03% Pepsin A-B: Letters indicate that there is a statistically significant difference (P<0.05) in the same sample in different concentrations of bile salts at the same time. Data regarding the physicochemical, microbiological and sensory analysis results of yogurt made with nanoencapsulated culture; Table 5. Yogurt production and incubation times with different ratios of nanoencapsulated starter. Incubation Time (Hour) Control 9.5 4.64±0.00 Table 6. pH values of yogurt samples. Storage Time of Yogurt Samples (Day) pH Value 4.12±0.028Aa 4.07±0.000Aab 4.06±0.028Abc 4.05±0.000Abc 4.01±0.028Ac 4.10±0.014Aa 4.08±0.000Aa 4.02±0.014Ab 4.08±0.014Aa 4.02±0.028Ab 4.12±0.014Aa 4.06±0.000Aa 4.08±0.056Aa 4.07±0.007Aa 4.07±0.028Aa 3.89±0.014Ba 3.81±0.014Bb 3.76±0.014BC 3.90±0.028Ba 3.90±0.014Ba A-B: Letters indicate that there is a statistically significant difference between the pH value in samples with different concentrations in the same storage period (P<0.05). a-C: Letters indicate that there is a statistically significant difference between the pH value in samples with the same concentration in different storage periods (P <0.05). Table 7. Acidity (% Lactic Acid) values of yogurt samples. Yogurt Samples Storage Period (Day) Lactic acid values (%) 1.09±0.003Cd 1.15±0.006BC 1.20±0.003Bb 1.24±0.011Ba 1.19±0.006C'° 1.07±0.004De 1.08±0.003Dd 1.18±0.004Cb 1.12±0.000Dc 1.22±0.003Ba 1.12±0.003Bdc 1.13±0.003CC 1.14±0.004Db 1.19±0.008ca 1.11±0.003Ddl 1.16±0.003Ad 1.27±0.001Ab 1.25±0.006AC 1.31±0.008Aa 1.31±0.003Aai A-D: Letters indicate that there is a statistically significant difference between the acidity value in samples with different concentrations in the same storage period (P <0.05). a-e: Letters indicate that there is a statistically significant difference between the acidity value in samples with the same concentration in different storage periods (P <0.05). Table 8. Water holding capacity of yogurt samples. Storage Period (Days) Water Retention Capacity (%) 84.16±0.056Bd 85.09±0.098BC 85.91±0.155Bb 86.58±0.000ABa 84.21±0.141Bd 85.44±0.155ABC 86.01±0.000Bb 87.03±0.028ABa 85.32±0.254Ab 85.59±0.000Ab 86.83±0.120Aab 88.04±1.442Aa Yogurt Samples A 72.39±0.028De B 74.20±0.141Be C 74.78±0.113^C D 73.60±0.141Cd 84.02±0.014BC 84.35±0.282CC 84.81±0.014Cb 85.95±0.042ca A-D: Letters indicate that there is a statistically significant difference between the water holding capacity values in samples with different concentrations in the same storage period (P<0.05). a-e: Letters indicate that there is a statistically significant difference between the water holding capacity values in samples with the same concentration in different storage periods (P<0.05). Table 9. Serum separation values in yogurt samples. Storage Period (Day) Serum Separation (ml/25 g) 4.17±0.028Bb 3.21±0.042BC 2.94±0.056Bd 2.85±0.070Ad 3.91±0.084Cb 3.11±0.070BC 2.86±0.056Bd 2.52±0.028Be 2.90±0.070Db 1.88±0.028CC 1.66±0.084Cd 1.57±0.028'3d Yogurt Samples A 4.94±0.028Ba B 4.71±0.014Ba C 3.77±0.098ca D 9.61±0.155Aa 8.35±0.141Ab .40±0.141AC 3.70±0.070Ad 2.32±0.028ce indicates that there is a statistically significant difference between the numbers of microorganisms in the sample with different concentrations within the same storage period (P<0.05). A-D: Letters indicate that there is a statistically significant difference between the numbers of microorganisms in the sample with different concentrations within the same storage period a-e: Letters indicate that there is a statistically significant difference between the numbers of serum separations in the sample with different concentrations within the same storage period (P<0.05). Table 10. Microbiological analysis results of yogurt samples. Yogurt Storage Samples Duration (Day) Str. thermophilus Lb. bulgaricus TAMBS A-D: Letters indicate that there is a statistically significant difference between the numbers of microorganisms in the sample with different concentrations within the same storage period (P<0.05). a-e: Letters indicate that there is a statistically significant difference between the numbers of microorganisms in samples with the same concentration at different storage times (P <0.05). Table 11. Sensory evaluation results of yogurt samples.TR TR TR TR TR TR TR TR TR

Claims (2)

1. The invention is related to the nanoencapsulated yogurt starter culture production method and its feature is; To obtain nanoencapsulation, mixing 0.1-5% Salep Glucomannan with 7-50% WPl, keeping this solution at room temperature in the laboratory for 2 hours and then mixing it at 500 rpm, centrifuging it at 1500 rpm for 20 minutes to separate the Salep glucomannans. adding 40% whey protein isolate (WPl) to the glucomannan solution obtained, mixing at 85-90 °C and 600 rpm for 30 minutes to ensure the denaturation of WPl. The number of bacteria in this prepared solution is 107-109. cfu/ml, and Lactobacillus delbrueckeii subsp. by electrospinning device with 0-30 kV and 0-1 mA sensitivity. Bulgaricus and Streptococcus are mixed at rpm and inoculated into a homogenized solution and nanoencapsulated yogurt starter culture bacteria. 2. The invention is related to the milk contained in the nanoencapsulated yogurt starter culture production method, and its feature is; - Nanoencapsulated starter culture ratio in 100ml in the entire formulation is 0.5-2%, v/v, - % total dry matter of the milk is 13.5%, - % water ratio of the milk is 84.5-86%.