CN117535182A - Low-post-acid direct-throw type yoghurt starter and preparation method thereof - Google Patents

Low-post-acid direct-throw type yoghurt starter and preparation method thereof Download PDF

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Publication number
CN117535182A
CN117535182A CN202311440890.3A CN202311440890A CN117535182A CN 117535182 A CN117535182 A CN 117535182A CN 202311440890 A CN202311440890 A CN 202311440890A CN 117535182 A CN117535182 A CN 117535182A
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streptococcus thermophilus
lactobacillus helveticus
acid direct
low
yoghurt
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马新
喻扬
郁雪平
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Thankcome Biotechnology Suzhou Co ltd
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Thankcome Biotechnology Suzhou Co ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/127—Fermented milk preparations; Treatment using microorganisms or enzymes using microorganisms of the genus lactobacteriaceae and other microorganisms or enzymes, e.g. kefir, koumiss
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/46—Streptococcus ; Enterococcus; Lactococcus

Abstract

The invention belongs to the field of food fermentation, and particularly relates to a low-post-acid direct-vat-set yoghurt starter and a preparation method thereof. The invention provides a low-post-acid direct-vat-set starter, which comprises streptococcus thermophilus NJ21 and lactobacillus helveticus LH-08B; the preservation number of the streptococcus thermophilus NJ21 is CCTCC No. M2014250, and the preservation number of the lactobacillus helveticus LH-08B is CCTCC No. M2019013. The invention prepares the direct-throwing yoghurt starter by compounding the two bacteria according to a certain proportion, is used as the starter to prepare the yoghurt, has high fermentation speed, and has weak acid and good quality after the prepared yoghurt.

Description

Low-post-acid direct-throw type yoghurt starter and preparation method thereof
Technical Field
The invention belongs to the field of food fermentation, and particularly relates to a low-post-acid direct-vat-set yoghurt starter and a preparation method thereof.
Background
The yoghurt is a sour and sweet milk beverage, and is a milk product which is prepared by taking milk as a raw material, pasteurizing, adding beneficial bacteria (a starter) into the milk, fermenting, and cooling and filling. Yogurt products in the market are mostly in the form of solidification, stirring and fruit flavor with various fruit jam and other auxiliary materials. The longer the yogurt is placed, the more sour the mouthfeel will be, but the lactic acid bacteria activity will be rather reduced. The phenomenon is acidification after the yoghurt, and the acidification after the yoghurt (post-acidification of yogurt) refers to the phenomenon that the residual lactobacillus in the yoghurt continuously generates acid by using lactose in the storage and transportation process after the yoghurt is fermented, so that the taste of the yoghurt is deteriorated. After the acidophilus milk is acidified, after the acidophilus milk is fermented normally, the thalli can still be fermented slowly by utilizing the residual lactose in the storage, transportation and sales processes, so that the lactic acid is continuously produced, and the acidity of the acidophilus milk is continuously increased. After accumulating to a certain extent, the generated peracid taste is unacceptable to consumers and loses value as a commodity.
In response to this problem, many scholars have proposed various measures to inhibit post-acidification, including refrigeration, use of additives, heat treatment, and altering the composition of the yogurt starter, among others. The two methods of rapid cooling after fermentation and low-temperature storage are widely accepted, and the problem of post acidification of the yoghourt in actual production of factories is solved to a certain extent, wherein the rapid cooling after fermentation is used for inhibiting the growth of strains, rapidly reducing the activity and reducing the acid production, thereby slowing down the acidification after the yoghourt, and the low-temperature storage is also the same reason, which is one of the most critical factors for ensuring the weak post acidification of the yoghourt. However, these measures have only been changed from the production process, and the post-acidification problem cannot be solved from the source and the production cost is increased.
Chinese patent CN 115948290A relates to a normal temperature weak post-acidification lactobacillus fermentum grx501 and application thereof, wherein the lactobacillus fermentum grx501 is preserved in the common microorganism center of the chinese microbiological bacterial culture collection center in 2022, 7 months and 20 days, and is classified and named as: lactobacillus fermentum (Lactobacillus fermentum) with a collection number of CGMCC No.25349. The lactobacillus fermentum grx501 was isolated from conventional fermented kimchi. By the invention, lactobacillus with weak growth ability and storage resistance in the milk environment is screened from non-dairy product environment (pickle); after the bacterial strain is identified, the bacterial strain is added into the sterilized yoghourt, so that the product still has weak post acidification under the normal temperature condition on the premise of maintaining higher viable count. After the yogurt is sterilized, lactobacillus fermentum grx501 is added, and the yogurt is stored for 21 days under the storage condition of 4 ℃ and 25 ℃, the pH of the sample is reduced by 0.02-4.08 and 0.03-4.10 respectively, the acidity is increased by 18.98 DEG T-103.19 DEG T and 20.33 DEG T-103.06 DEG T respectively, and the yogurt still has weak post acidification at normal temperature. Can be applied to the production of the acidulated viable bacteria type fermented milk after weak at normal temperature.
Chinese patent CN 116694539A discloses a lactobacillus plantarum J26 as a direct vat set starter, and a preparation method and application thereof, wherein the preparation method comprises the following steps: lactobacillus plantarum J26 is activated, inoculated into 100mL of optimized carbon-nitrogen source culture medium according to the inoculation amount of 4 percent, subjected to high-density fermentation by adopting a 5L fermentation tank for 24 hours at the temperature of 33 ℃ and the pH value of 5.6, fed at a constant speed of 4-24 hours, wherein the feeding rate is 15mL/h, and the fed culture medium is the optimized carbon source: optimized nitrogen source = 5:2; and after the high-density fermentation is finished, the bacterial liquid is harvested and freeze-dried to obtain freeze-dried bacterial powder. The combination of the added compound prebiotics and the lactobacillus plantarum J26 serving as the direct vat set starter can promote the fermentation of the soybean milk, remarkably improve the viable count of the fermented soybean milk and slow down the post acidification of the fermented milk.
In order to meet the increasing quality requirements of yogurt and dairy products, more low-post-acid direct-vat-set yogurt starter needs to be developed, the yogurt fermentation step is simplified, the production efficiency is improved, the production cost is controlled, the yogurt has better taste and high probiotics content.
Disclosure of Invention
In order to solve the problems, the invention provides a low-post-acid direct-vat-set starter, which comprises streptococcus thermophilus NJ21 and lactobacillus helveticus LH-08B, wherein the ratio of the viable bacteria number of the streptococcus thermophilus NJ21 to the lactobacillus helveticus LH-08B is 5-14:15-6.
In one aspect, the invention provides a low post-acid direct-vat-set starter comprising streptococcus thermophilus NJ21 and lactobacillus helveticus LH-08B; the preservation number of the streptococcus thermophilus NJ21 is CCTCCNO. M2014250; the preservation number of the Lactobacillus helveticus LH-08B is CCTCC NO. M2019013.
Specifically, the ratio of the viable count of streptococcus thermophilus NJ21 to the viable count of lactobacillus helveticus LH-08B is 5-14:15-6.
In one embodiment, the ratio of the number of viable bacteria of Streptococcus thermophilus NJ21 to Lactobacillus helveticus LH-08B is 5:15.
In another embodiment, the ratio of the number of viable bacteria of Streptococcus thermophilus NJ21 to the number of viable bacteria of Lactobacillus helveticus LH-08B is 1:1.
In another example, the ratio of viable bacteria of Streptococcus thermophilus NJ21 to Lactobacillus helveticus LH-08B is 14:6.
Specifically, the total number of added live bacteria of streptococcus thermophilus NJ21 and lactobacillus helveticus LH-08B can be 10 5 -10 6 cfu/mL。
In yet another aspect, the present invention provides a method for preparing the low post acid direct vat set starter, comprising the steps of:
(1) Culturing streptococcus thermophilus NJ21 and lactobacillus helveticus LH-08B respectively to obtain seed liquid;
(2) Inoculating the seed liquid obtained in the step (2) into a fermentation tank for fermentation respectively according to the inoculum size of 5-10%, so as to obtain fermentation liquid;
(3) Respectively centrifuging the fermentation liquor obtained in the step (2) to obtain bacterial sludge;
(4) Uniformly mixing the bacterial mud obtained in the step (3) with a freeze-drying protective agent respectively, and freeze-drying to obtain freeze-dried powder;
(5) And (3) mixing the freeze-dried powder obtained in the step (4) to obtain the low-post-acid direct-vat set yoghurt starter.
Specifically, the lyoprotectant in the step (3) is: according to weight percentage, trehalose 5-15%, skim milk powder 10-15%, lactose 4-8%, inulin 4-8%, glycerin 2-5%, mannose 2-5%, sodium glutamate 3-6%, polyethylene glycol 1-3%, and the balance water.
Preferably, the lyoprotectant of step (3) is: according to weight percentage, trehalose is 10%, skim milk powder is 15%, lactose is 4%, inulin is 6%, glycerin is 2%, mannose is 5%, sodium glutamate is 6%, polyethylene glycol is 3%, and the balance is water.
Specifically, the bacterial sludge in the step (3) and the protective agent solution are mixed in a mass ratio of 1:3-1:5.
Specifically, the mixing proportion of the freeze-dried powder in the step (4) is that the viable count ratio of streptococcus thermophilus NJ21 to lactobacillus helveticus LH-08B is 5-14:15-6.
In one embodiment, the ratio of the number of viable bacteria of Streptococcus thermophilus NJ21 to Lactobacillus helveticus LH-08B is 5:15.
In another embodiment, the ratio of the number of viable bacteria of Streptococcus thermophilus NJ21 to the number of viable bacteria of Lactobacillus helveticus LH-08B is 1:1.
In another embodiment, the ratio of viable bacteria of Streptococcus thermophilus NJ21 to Lactobacillus helveticus LH-08B is 14:6.
In yet another aspect, the invention provides a yogurt production method comprising using the low post acid direct vat set yogurt starter.
Specifically, the total number of added viable bacteria of the low-post-acid direct-vat-set starter is 10 5 -10 6 cfu/mL。
Specifically, the fermentation temperature is 35-40 ℃, and the fermentation is carried out for 8-15 hours.
Preferably, the fermentation temperature is 37 ℃, and the fermentation is carried out for 10 hours.
In particular, fermentation bases, sweeteners and/or stabilizers are also included.
Specifically, the sweeteners include, but are not limited to: white granulated sugar B, glucose, fructose syrup, erythritol, mannitol, maltitol, xylitol, isomalt, isomaltulose, lactitol, sorbitol, galacto-oligosaccharides, isomalt-oligosaccharides, polydextrose, fructo-oligosaccharides, xylo-oligosaccharides, soy oligosaccharides, aspartame, acesulfame potassium, sucralose, stevioside, lo Han Guo extract and thaumatin.
Specifically, the stabilizers include, but are not limited to: starch, pectin, soybean polysaccharide, gelatin, agar, etc.
In yet another aspect, the invention provides the use of the low post acid direct vat set starter described above in the production of fermented dairy products.
Specifically, the fermented dairy product includes, but is not limited to: soy milk, normal temperature yoghurt, low temperature yoghurt, stirred yoghurt, set yoghurt, drinkable yoghurt, cheese or lactobacillus beverage.
The invention has the technical effects that:
(1) The viable count of lactobacillus in the yogurt is high, and after 21d of refrigeration, the viable count is 10 6 The above;
(2) The direct-vat-set yoghurt starter can effectively inhibit acidification after the yoghurt so that the acidity change of the yoghurt in the refrigeration 21d is small and is always maintained within 119 DEG T.
Detailed Description
The present invention will be described in further detail with reference to the following examples, which are not intended to limit the present invention, but are merely illustrative of the present invention. The experimental methods used in the following examples are not specifically described, but the experimental methods in which specific conditions are not specified in the examples are generally carried out under conventional conditions, and the materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.
The terms "include" or "comprising" as used herein mean "including but not limited to". The term is intended to be open ended to specify the presence of any stated features, elements, integers, steps, or components, but does not preclude the presence or addition of one or more other features, elements, integers, steps, components, or groups thereof. Thus, the term "comprising" includes the more limiting terms "consisting of … …" and "consisting essentially of … …". In one embodiment, the term "comprising" as used throughout the application, and in particular in the claims, may be replaced by the term "consisting of … …".
The terms "optional," "any," or "any" used herein mean that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "optionally comprising 1 antibody heavy chain variable region" means that an antibody heavy chain variable region of a particular sequence may be, but is not required to be, present.
The term "and/or" as used herein is understood to mean any one of the selectable items or a combination of any two or more of the selectable items.
Example 1
1.1 Strain
Streptococcus thermophilus (Streptococcus thermophilus) NJ21 with a preservation number of CCTCCNO. M2014250;
lactobacillus helveticus (Lactobacillus helveticus) LH-08B with the preservation number of CCTCC No. M2019013;
streptococcus thermophilus JCCC 0033 with the preservation number of CGMCC NO.22560;
lactobacillus helveticus CCFM1034 with deposit number GDMCC No.60451.
1.2 preparation of direct-throw yoghurt starter
(1) Culturing the strain. Respectively inoculating NJ21 and LH-08B into liquid culture medium in a triangular flask for culturing at 35-40deg.C for 24-48 hr to obtain seed solution, and respectively inoculating into a fermenter for fermentation culture at 35-40deg.C for 24-48 hr with 5-10% of inoculum size.
The formula of the streptococcus thermophilus culture medium is as follows: 5.0g/L of soybean peptone, 2.5g/L of casein peptone, 2.5g/L of yeast extract powder, 5.0g/L of beef extract powder, 5.0g/L of lactose, 0.5g/L of sodium ascorbate, 19.0g/L of sodium B-glycerophosphate, 0.25g/L of magnesium sulfate and pH of 6.3.
The formula of the lactobacillus helveticus culture medium is as follows: 10g/L casein peptone, 5g/L yeast extract powder, 10g/L beef extract, 20g/L glucose, 2g/L diamine citrate, 2g/L dipotassium hydrogen phosphate, 5g/L sodium acetate and MgSO 4 ·7HO0.58g/L，MnSO 4 ·4H 2 O0.25 g/L, tween-80 1mL/L, L-cysteine 0.5g/L, pH 7.0.
(2) And (5) preparing freeze-dried powder. Respectively centrifuging the fermentation liquor obtained in the step (1) to obtain bacterial sludge; and (3) uniformly mixing the bacterial sludge with a freeze-drying protective agent 1:1 (w/w), and freeze-drying to obtain NJ21 and LH-08B freeze-dried powder, and respectively detecting the viable count of the bacterial sludge.
The formula of the freeze-drying protective agent is as follows: according to weight percentage, trehalose is 10%, skim milk powder is 15%, lactose is 4%, inulin is 6%, glycerin is 2%, mannose is 5%, sodium glutamate is 6%, polyethylene glycol is 3%, and the balance is water.
(3) And (3) preparing the direct-throwing yoghurt starter. Mixing the two freeze-dried powders in the step (2) according to a proportion, wherein the ratio of the viable bacteria number of the NJ21 to the viable bacteria number of the LH-08B is 25:75.
1.3 yogurt fermentation Process
Sterilizing fresh milk at 95deg.C for 20min or at 140deg.C for 2s, cooling to 30deg.C, adding direct-feeding yogurt starter, and adding 10% of the additive 5 -10 6 cfu/mL (preferably 10) 6 cfu/mL), fermentation at 37℃for 8-15h, pH 4.4, and termination. And (5) refrigerating the fermented yoghourt at 4 ℃.
Example 2
The difference from example 1 is that the ratio of the total number of viable bacteria of NJ21 and LH-08B is 50:50.
Example 3
The difference from example 1 is that the ratio of the total number of viable bacteria of NJ21 and LH-08B is 70:30.
Comparative example 1
The difference from example 1 is that NJ21 was replaced with JMCC0033.
Comparative example 2
The difference from example 1 is that LH-08B is replaced with CCFM1034.
Comparative example 3
The difference from example 1 is that only NJ21 lyophilized powder was used.
Comparative example 4
The difference from example 1 is that only LH-08B lyophilized powder is used.
Example 5 evaluation of preparation of yogurt by direct-throw yogurt starter
5.1 total number of viable lactic acid bacteria in yogurt
The total number of viable lactic acid bacteria in the yogurt of examples 1 to 4 and comparative examples 1 to 4 was examined according to GB 4789.35-2010 food microbiology test lactic acid bacteria test standard, and the examination results are shown in Table 1.
TABLE 1
As is clear from Table 1, the total viable count of lactic acid bacteria in examples 1 to 4 remains 10 after 21d of cold storage at 4 ℃ 6 The above meets the specification of the national standard GB 19302-2010; comparative examples 1-4 the total viable count of lactic acid bacteria after 21d was one order of magnitude lower than the national standard.
5.2 post-yoghurt acidification detection
Examples 1-4 the yoghurt of comparative examples 1-4 was refrigerated at 4 ℃ for 21d and the acidity change was recorded and the results of the measurements are shown in table 2.
TABLE 2
As is clear from Table 2, the acidity of examples 1-4 in the refrigerated storage 21d is not greatly changed and is always maintained within 119 DEG T, so that the yogurt keeps good taste, and the ratio of the viable count of Streptococcus thermophilus NJ21 and Lactobacillus helveticus LH-08B in the examples can effectively inhibit post-acidification phenomenon of the yogurt when the yogurt is used for fermentation, thereby remarkably improving the shelf life and sensory characteristics of the yogurt product and improving the quality of the yogurt product.

Claims (10)

1. The low-post-acid direct-throw type yoghurt starter is characterized by comprising streptococcus thermophilus NJ21 and lactobacillus helveticus LH-08B; the preservation number of the streptococcus thermophilus NJ21 is CCTCCNO. M2014250; the preservation number of the Lactobacillus helveticus LH-08B is CCTCC NO. M2019013.
2. The low post acid direct vat set starter according to claim 1, wherein said live bacteria count ratio of streptococcus thermophilus NJ21 to lactobacillus helveticus LH-08B is 5-14:15-6.
3. The low post acid direct vat set starter according to claim 2, wherein said live bacteria count ratio of streptococcus thermophilus NJ21 to lactobacillus helveticus LH-08B is 5:15.
4. The low post acid direct vat set starter according to claim 2, wherein said live bacteria count ratio of streptococcus thermophilus NJ21 to lactobacillus helveticus LH-08B is 1:1.
5. The low post acid direct vat set starter according to claim 2, wherein said live bacteria count ratio of streptococcus thermophilus NJ21 to lactobacillus helveticus LH-08B is 14:6.
6. The low post acid direct vat set starter according to any one of claims 1 to 5, wherein said streptococcus thermophilus NJ21 and lactobacillus helveticus LH-08B add viable bacteria in total 10 5 -10 6 cfu/mL。
7. A method for preparing the low post acid direct vat set yoghurt starter according to any one of claims 1 to 6, comprising the steps of:
(1) Culturing streptococcus thermophilus NJ21 and lactobacillus helveticus LH-08B respectively to obtain seed liquid;
(2) Inoculating the seed liquid obtained in the step (2) into a fermentation tank for fermentation respectively according to the inoculum size of 5-10%, so as to obtain fermentation liquid;
(3) Respectively centrifuging the fermentation liquor obtained in the step (2) to obtain bacterial sludge;
(4) Uniformly mixing the bacterial mud obtained in the step (3) with a freeze-drying protective agent respectively, and freeze-drying to obtain freeze-dried powder;
(5) And (3) mixing the freeze-dried powder obtained in the step (4) to obtain the low-post-acid direct-vat set yoghurt starter.
8. The method of claim 7, wherein the lyoprotectant of step (3) is: according to weight percentage, trehalose 5-15%, skim milk powder 10-15%, lactose 4-8%, inulin 4-8%, glycerin 2-5%, mannose 2-5%, sodium glutamate 3-6%, polyethylene glycol 1-3%, and the balance water.
9. The method of claim 8, wherein the bacterial sludge of step (3) is mixed with the protectant solution in a mass ratio of 1:3 to 1:5.
10. A method for preparing yoghurt, comprising the use of a low post acid direct vat set yoghurt starter according to any one of claims 1 to 6.
CN202311440890.3A 2023-11-01 2023-11-01 Low-post-acid direct-throw type yoghurt starter and preparation method thereof Pending CN117535182A (en)

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