CN115505539B - Kluyveromyces marxianus and application thereof in fermented food - Google Patents

Kluyveromyces marxianus and application thereof in fermented food Download PDF

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CN115505539B
CN115505539B CN202211217136.9A CN202211217136A CN115505539B CN 115505539 B CN115505539 B CN 115505539B CN 202211217136 A CN202211217136 A CN 202211217136A CN 115505539 B CN115505539 B CN 115505539B
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fermented
kluyveromyces marxianus
yjmk
milk
whey
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CN115505539A (en
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李妍
张列兵
李扬
杨茜茜
朱园花
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Beijing Technology and Business University
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Abstract

The invention relates to the technical field of food processing, in particular to Kluyveromyces marxianus and application thereof in fermented food. The invention discloses Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 which is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of 25655. The Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 is inoculated into cow milk after being subjected to expansion culture or is compounded with lactobacillus to be inoculated into cow milk, so that cow milk can be endowed with a brake taste and a unique flavor.

Description

Kluyveromyces marxianus and application thereof in fermented food
Technical Field
The invention relates to the technical field of food processing, in particular to Kluyveromyces marxianus and application thereof in fermented food.
Background
Kluyveromyces marxianus belongs to Kluyveromyces, and is listed as edible fungus, and can be added into fermented food. Patent CN201510527940.0 discloses a new kluyveromyces marxianus strain which can be used for preparing black tea fungus protein beverage and lactose hydrolysis. Research has also shown that when Kluyveromyces marxianus as a milk beer fermentation strain is used, the strain can partially or completely replace beer yeast or Saccharomyces cerevisiae to produce milk beer. Patent CN201810186371.1 discloses a freeze-dried milk beer starter compounded by Kluyveromyces marxianus and Saccharomyces cerevisiae. Patent CN201710355851.1 discloses a preparation method and application of Kluyveromyces marxianus freeze-dried bacterial agent. The above studies indicate that the use of Kluyveromyces marxianus and other species to compound post-fermented beverages is technically feasible, but the influence of Kluyveromyces marxianus on milk flavor and organoleptic properties is not disclosed in the above documents. The prior art lacks Kluyveromyces marxianus capable of improving flavor substances and organoleptic properties of fermented beverage.
Disclosure of Invention
The invention provides Kluyveromyces marxianus and application thereof in fermented food, which can be used for producing fermented beverage with unique taste and flavor characteristics and enriching fermented beverage products.
The invention provides Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 which is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of 25655. Preservation date: 2022, 9, 7; deposit unit address: beijing, chaoyang area, north Chen Xi Lu No. 1, 3, china academy of sciences microbiological institute, post code: 100101.
the invention also provides a microbial inoculum, which comprises the Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3.
According to the microbial inoculum provided by the invention, the microbial inoculum also comprises lactic acid bacteria.
In some embodiments, the ratio of live bacteria count of kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 to lactic acid bacteria is: 3:1 to 1:5.
according to the microbial inoculum provided by the invention, the lactobacillus is any one or more of lactobacillus bulgaricus, streptococcus thermophilus, lactococcus lactis and leuconostoc mesenteroides.
Preferably, the lactobacillus is lactobacillus bulgaricus or streptococcus thermophilus.
Further preferably, the ratio of the number of viable bacteria of the lactobacillus bulgaricus to the streptococcus thermophilus is 1: 5-5: 1.
the invention also provides a fermentation product which is prepared by fermenting and culturing the Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 or the microbial inoculum.
The fermentation product provided by the invention is fermented milk, fermented whey or gas-containing fermented beverage.
The gas-containing fermented beverage is prepared from fermented milk or fermented whey.
The invention also provides a culture medium of Kluyveromyces marxianus, which comprises the following components: glucose 35-45g/L, yeast extract 2.5-5.0g/L, soytone 10-15g/L. The culture medium can be used for the YJMK-3 amplification culture.
Preferably, the pH of the medium is adjusted to 6.5.+ -. 0.2.
In some embodiments, after the Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 is amplified and cultured, the viable count of the obtained culture solution is 1-7 multiplied by 10 8 CFU/mL. The organoleptic properties and the like of cow milk can be improved by utilizing the strain or the strain and lactic acid bacteria to ferment cow milk together.
The conditions for the preferred expansion culture are: culturing at 30+/-2 deg.c (30 deg.c preferably) for 10-16 hr; further preferably, air culture is performed.
Preferably, after the expansion culture, the number of YJMK-3 viable bacteria is 1-7×10 8 CFU/mL。
Preferably, kluyveromyces marxianus is cultured until the viable count is 5×10 8 CFU/mL, the number of viable bacteria cultivated by the culture medium of the invention is increased by more than two times compared with YPD culture medium.
The invention also provides a preparation method of the fermented milk, which comprises the following steps: after the raw milk is sterilized, bacteria are added for fermentation, wherein the bacteria are YJMK-3 yeast or YJMK-3 yeast and lactobacillus (preferably lactobacillus bulgaricus and streptococcus thermophilus are compounded).
Preferably, the preparation method of the fermented milk comprises the following steps: sterilizing raw milk, and adding the microbial inoculum for fermentation.
In some embodiments, the inoculum size of the bacterium is 2% -6%.
In some embodiments, preparing the kefir fermented milk comprises the steps of:
1) Preparing raw milk:
raw milk may be defatted, partially defatted or non-defatted using fresh cow milk;
the raw milk can be reconstituted milk with a solid content of 9.5% -11%. Preparing reconstituted milk: adding water into milk powder raw materials such as skimmed milk powder or whole milk or milk protein powder, and stirring thoroughly for dissolving; the hydration condition is that the hydration is carried out for 0.5 to 1 hour at 45 to 50 ℃ and/or 8 to 12 hours below 6 ℃;
2) Sterilizing the raw milk at 85-95deg.C for 5-15min.
3) Adding YJKM-3 yeast into the sterilized raw milk obtained in the step 2), wherein the inoculation amount is 2% -6%; or YJKM-3 yeast and lactobacillus (one or more of Lactobacillus bulgaricus, streptococcus thermophilus, lactococcus lactis and Leuconostoc mesenteroides) are inoculated, wherein the inoculation ratio of the yeast and the lactobacillus is 3:1 to 1:5 (preferably 1:1).
4) Fermenting the sample prepared in the step 3) at 30+/-5 ℃ for 12-36 hours, or fermenting at 40-42 ℃ for 4-6 hours, cooling to 30+/-5 ℃ and continuing fermenting for 12-24 hours. Stirring is stopped during fermentation, and then cooling and storing are carried out.
Preferably, the preparation method of the fermented milk comprises the following steps:
1) Preparing raw milk: taking reconstituted cow milk with the solid content of 9.5% -11%, and shearing for 0.5-5min at 5000-10000 rpm; the hydration condition is 45-50 ℃, the hydration is carried out for 0.5-1h, or the hydration is carried out for 8-12 h below 6 ℃; or directly taking fresh cow milk as raw milk.
2) Sterilizing the raw milk at 85-95deg.C for 5-15min.
3) Adding Kluyveromyces marxianus into the sterilized raw milk obtained in the step 2), wherein the inoculation amount is 2% -6%; or the Kluyveromyces marxianus is inoculated with lactobacillus (lactobacillus bulgaricus and streptococcus thermophilus) in a compounding way, and the inoculation ratio of the saccharomycetes to the lactobacillus is 3:1 to 1:5 (preferably 1:1).
4) And (3) fermenting the sample prepared in the step (3) at 40-42 ℃ for 4-6 hours, cooling to 30-35 ℃, continuing fermenting for 12-30 hours, and cooling and storing after finishing fermentation.
The invention also provides a preparation method of the fermented whey, which comprises the following steps: after the raw material whey liquid is sterilized, bacteria are added for fermentation, wherein the bacteria are YJMK-3 Kluyveromyces marxianus or YJMM-3 Kluyveromyces marxianus and lactobacillus (preferably lactobacillus bulgaricus and streptococcus thermophilus are compounded).
Preferably, the preparation method of the fermented whey comprises the following steps: sterilizing raw whey, and adding the microbial inoculum for fermentation.
In some embodiments, the inoculum size of the bacterium is 2% -6%.
In some embodiments, preparing the kefir marxianus fermented whey comprises the steps of:
1) Preparation of raw whey:
fresh or concentrated whey can be used as raw whey;
the raw material whey can be recovered whey with the solid content of 10% -25%, and the recovered whey is prepared by the following steps: adding water into whey products such as whey powder, desalted whey powder and/or whey protein, and stirring thoroughly for dissolving; the hydration condition is that the hydration is carried out for 0.5 to 1 hour at 45 to 50 ℃ and/or 8 to 12 hours below 6 ℃;
2) Sterilizing the raw material whey at 85-95deg.C for 5-15min.
3) Adding YJKM-3 Kluyveromyces marxianus into the sterilized raw material whey in the step 2), wherein the inoculation amount is 2% -6%; or YJKM-3 Kluyveromyces marxianus and lactobacillus (one or more of Lactobacillus bulgaricus, streptococcus thermophilus, lactococcus lactis and Leuconostoc mesenteroides) are combined and inoculated, and the inoculation ratio of the saccharomycetes to the lactobacillus is 3:1 to 1:5 (preferably 1:1).
4) Fermenting the sample prepared in the step 3) at 30+/-2 ℃ for 12-36 hours, or fermenting at 40-42 ℃ for 4-6 hours, cooling to 30+/-2 ℃ and continuing fermenting for 12-24 hours, intermittently stirring during the fermentation period, and then cooling and storing.
In some embodiments, the method of preparing an aerated fermented beverage comprises: the fermented milk or fermented whey is used as raw materials, added with water, an acidulant, a thickening emulsifier, and optionally added with sweeteners, fruit and vegetable juice, essence and the like, and is prepared by blending, homogenizing and sterilizing.
Preferably, the fermented milk (5% -40%) or the fermented whey (5% -40%) is added, water, an acidulant, a thickening emulsifier and optionally a sweetener, fruit and vegetable juice, essence and the like are added, and the fermented milk is prepared by blending, homogenizing and sterilizing. Optionally filling or not filling carbon dioxide gas in the processing process, and sterilizing again after filling.
The sour agent is one or more of citric acid, malic acid, sodium citrate and lactic acid.
The thickening emulsifier is one or more of CMC, pectin, PGA, sodium tripolyphosphate, gellan gum, polyglycerol fatty acid ester and xanthan gum.
The sweetener is one or more of white granulated sugar, high fructose syrup, sodium cyclamate, aspartame, stevioside, sucralose, erythritol, sugar alcohols and the like.
The fermented product prepared by the preparation method has unique taste and flavor characteristics.
The application of the Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 or the microbial inoculum in any one of the following:
1) The taste of the fermented beverage is improved;
2) Adding flavor substances to the fermented product;
3) The mellow flavor of the fermented product is improved;
4) Improving the quality of fermented milk, fermented whey or gas-containing fermented beverage;
5) Promoting the metabolism of protein by lactic acid bacteria;
6) Improving the milk metabolism ability;
7) Provides a fermented beverage with unique flavor.
According to the application, the flavor substance comprises one or more of alcohols, methyl ketones, ethyl esters and acids.
According to the above application, the flavoring substances comprise one or more of ethanol, isoamyl alcohol, 2-heptanol, 2-nonanone, 2-undecanone, ethyl caproate, ethyl caprylate, ethyl caprate, acetic acid, caproic acid, caprylic acid, capric acid, benzaldehyde and delta decalactone.
The invention has the beneficial effects that:
the invention screens a strain of Kluyveromyces marxianus from the kefir yoghurt which is naturally fermented, expands the culture and is used for fermenting dairy products, and the Kluyveromyces marxianus has obvious influence on the sensory and flavor compound composition of the fermented dairy products, and can be applied to the processing of the fermented dairy products.
The invention provides Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 and application thereof in fermented foods, and the yeast is inoculated into cow milk after being subjected to expansion culture or is compounded with lactobacillus to be inoculated into cow milk, so that cow milk can be endowed with a brake taste and a unique flavor.
The cow milk obtained by metabolism of saccharomycetes contains 8 flavor compounds including 3 alcohols (ethanol, isoamyl alcohol and 2-heptanol), 2 methyl ketones (2-nonone and 2-undecone) and 3 ethyl esters (ethyl caproate, ethyl caprylate and ethyl caprate), and simultaneously gives the cow milk a brake taste.
After being compounded with lactic acid bacteria, YJMK-3 ferments cow milk, and the obtained cow milk contains 14 flavor substances, and besides the flavor compounds, 4 kinds of acids (acetic acid, caproic acid, caprylic acid and capric acid), benzaldehyde and delta decalactone are produced, so that the cow milk has more abundant flavor substances and obvious stopping taste.
Drawings
In order to more clearly illustrate the invention or the technical solutions of the prior art, the following description will briefly explain the drawings used in the embodiments or the description of the prior art, and it is obvious that the drawings in the following description are some embodiments of the invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the effect of soybean peptone concentration on proliferation of yeast YJMK-3 in example 1 of the present invention;
FIG. 2 is a graph showing the effect of glucose concentration on proliferation of yeast YJMK-3 in example 1 of the present invention;
FIG. 3 is a graph showing the effect of yeast extract concentration on proliferation of yeast YJMK-3 in example 1 of the present invention;
FIG. 4 is a validation chart of the orthogonal experiment optimized medium of example 1 of the present invention, notes: 1-9 corresponds to 9 sets of orthogonal experiments, 10 is a preferred medium validation plot;
FIG. 5 is the sensory evaluation results of the fermented cow's milk of comparative examples 1, 2 and example 2 of the present invention;
FIG. 6 shows the results of sensory evaluation of the fermented cow's milk of comparative examples 3 and 4 and example 4.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings, and it is apparent that the described embodiments are some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The experimental methods and conditions in the examples of the present invention are conventional methods unless otherwise specified. The reagents or materials, unless otherwise specified, are commercially available.
Example 1: enlarged culture of Kluyveromyces marxianus YJMK-3
1.1 screening of Kluyveromyces marxianus YJMK-3 Medium
The effect of soybean peptone concentration on yeast YJMK-3 proliferation is shown in FIG. 1, the effect of glucose concentration on yeast YJMK-3 proliferation is shown in FIG. 2, and the effect of yeast extract concentration on yeast YJMK-3 proliferation is shown in FIG. 3.
As shown in FIG. 4, the optimal combination of soybean peptone, yeast extract and glucose obtained by optimized screening of three-factor three-level orthogonal experiments is that the addition amounts of soybean peptone, yeast extract and glucose are respectively 10g/L, 2.5g/L and 35g/L. The number of the preferred culture medium bacteria is proved to be significantly higher than that of 9 groups of orthogonal experiment results.
1.2 Effect of culture time on the culture Effect of Kluyveromyces marxianus YJMK-3
Preferred Medium compared with conventional YPD Medium
1.3 expanded culture of Kluyveromyces marxianus YJMK-3
The preferred medium and YPD medium were prepared and sterilized at 121℃for 20 min. And (3) absorbing a proper amount of frozen bacterial liquid, placing the frozen bacterial liquid into a YPD liquid culture medium, culturing the frozen bacterial liquid in a constant temperature incubator at 30 ℃ for 12h for activation, continuously carrying out passage activation for 2 generations, inoculating the frozen bacterial liquid into a preferred culture medium, and culturing the frozen bacterial liquid at the constant temperature of 30 ℃ for 14h by aeration culture to obtain the YJMK-3 enlarged culture liquid. The number of viable bacteria was measured by sampling with YPD agar medium.
The preferred medium formulation is as follows:
composition of the components Content (g/L)
Peptone 10.0
Glucose 35.0
Yeast extract powder 2.5
pH 6.5±0.2
Example 2: application of Kluyveromyces marxianus YJMK-3 in fresh cow milk fermentation
(1) Sterilizing whole milk at 95deg.C for 5min;
(2) Adding an enlarged culture solution of Kluyveromyces marxianus into the sterilized whole milk obtained in the step (1), wherein the inoculation amount is 4%;
(3) And (3) demulsification is carried out on the to-be-fermented cow milk obtained in the step (2) at the constant temperature of 30 ℃ for 16-24 h and the rotating speed of 200rpm for 3 minutes, and refrigeration is carried out.
Example 3: application of Kluyveromyces marxianus YJMK3 in fermentation of reconstituted cow milk
(1) Rehydrating the whole milk powder, and stirring and dissolving the whole milk powder with drinking water which is 9 times of the weight of the milk powder;
(2) Dispersing the incompletely hydrated reconstituted milk obtained in the step (1) for 1min under the condition of 10000 rpm;
(3) Hydrating the dispersed reconstituted milk obtained in the step (2) in a water bath at 50 ℃ for 30min;
(4) Sterilizing the reconstituted milk obtained in the step (3) at 90 ℃ for 5min;
(5) And (3) adding the Kluyveromyces marxianus culture solution after the expansion culture into the sterilized reconstituted milk in the step (4), wherein the inoculation amount is 3%.
(6) Fermenting the reconstituted milk to be fermented in the step (5) at a constant temperature of 32 ℃ for 24 hours, stirring to demulsify, and refrigerating for 12 hours to obtain the fermented milk.
Example 4: application of Kluyveromyces marxianus YJMK3 and lactobacillus compound in fermented cow milk
(1) Sterilizing whole milk at 90deg.C for 5min;
(2) Adding the Kluyveromyces marxianus culture solution after the expansion culture into the sterilized whole milk in the step (1), wherein the inoculation amount is 2%; adding mixed bacterial liquid (the ratio of viable bacteria number is 1:1) of lactobacillus bulgaricus (LB 1) and streptococcus thermophilus (S10) 2%
(3) And (3) demulsification is carried out on the to-be-fermented cow milk obtained in the step (2) at the constant temperature of 30 ℃ for 16h and the rotating speed of 200rpm for 3 minutes, and refrigeration is carried out.
Example 5: fermented whey
Rehydrating whey powder, and dissolving with 8 times of water under stirring
Placing the stirred and dissolved whey powder in 5 ℃ heat-preserving water for 1h to obtain reconstituted whey
2) Sterilizing the raw material whey at 95deg.C for 5min.
3) Adding YJKM-3 Kluyveromyces marxianus into the sterilized raw material whey in the step 2), wherein the inoculation amount is 2%; adding mixed bacterial liquid (the ratio of viable bacteria number is 1:1) of lactobacillus bulgaricus (LB 1) and streptococcus thermophilus (S10) 2%
4) Fermenting the sample prepared in the step 3) at 42 ℃ for 4 hours, cooling to 30 ℃ and continuing fermenting for 14 hours, and stirring intermittently during the fermentation period, and then cooling and storing.
Example 6: application of Kluyveromyces marxianus YJMK3 in gas-containing fermented beverage
20% of fermented milk, 10% of sucrose and citric acid are added to adjust the acidity to pH4.2, CMC and pectin accounting for 0.4% of the total amount are added, after blending, homogenizing and sterilizing, carbon dioxide gas is filled, the fermented milk is filled into a pop can package, and the primary gas-containing fermented beverage is obtained after the secondary sterilizing treatment.
Comparative example 1
Comparative example 1 was a fermented milk obtained in the procedure of example 2 using conventional Kluyveromyces marxianus YJMK1 (YJMK 1 is a different strain of Kluyveromyces marxianus isolated from kefir).
Comparative example 2
Comparative example 2 was fermented cow's milk by the procedure of example 2 using Saccharomyces cerevisiae NJM1 (NJM 1 is a strain of Saccharomyces cerevisiae isolated from fermented horse milk).
Comparative example 3
Comparative example 3 was prepared by co-fermenting milk in accordance with example 4 using LB1 and S10 (the ratio of viable bacteria to each other is 1:1).
Comparative example 4
Comparative example 4 was prepared by co-fermenting cow' S milk in accordance with example 4 using conventional Kluyveromyces marxianus (YJMK 1) and LB1, S10 (viable count ratio of 1:1).
Table 1 comparative examples and examples 2, 4 viscosity, alcohol content, pH and total free amino acid concentration of fermented cow milk
As can be seen from Table 1, the total free amino acids of the fermented cow's milk of example 2 (YJMK-3 group) was 1.23mmoL/L, and the viscosity was 3.0cP, which were higher than those of comparative examples 1 and 2 under the same conditions. The alcohol content of example 2 was 0.46% higher than that of comparative example 1 but lower than that of comparative example 2. The above data indicate that the ability of example 2 (YJMK-3) to metabolize cow's milk is significantly higher than that of YJM 1 and NJM1 under the same conditions.
The total free amino acids of the fermented milk of the LB1+S10 group (comparative example 3) were 1.96mmoL/L, the alcohol content was 0.00%, and the viscosity was 694cP. The total free amino acids of the YJMK1+LB1+S10 group (comparative example 4) were 1.83mmoL/L similarly to the LB1+S10 group. The alcohol content was 0.27%, lower than that at the time of single-bacteria fermentation (0.35%), and the viscosity was 682cP. It can be seen that when YJMK1 is fermented together with two strains of lactic acid bacteria, the alcohol production capacity is reduced, and the total free amino acid concentration is not obviously increased or even reduced, which indicates that YJMK1 cannot promote the capacity of lactic acid bacteria to metabolize proteins. The total free amino acids of the fermented milk of YJMK-3+LB1+S10 group (example 4) were higher than those of the Lb1+S10 group and were 2.12mmoL/L. The alcohol content was 0.46%, consistent with that of the single-bacteria fermentation (0.46%), the viscosity was 758cP, and slightly higher than that of the LB1+S10 group. It can be seen that there is an interaction between YJMK-3 and two strains of lactic acid bacteria during co-fermentation, resulting in an increased degree of cow's milk proteolysis.
Table 2 comparative example flavor compound composition of fermented cow milk with examples 2 and 4
As can be seen from Table 2, the flavor compounds of comparative example 1 (YJMK 1) and example 2 (YJMM 3) are similar in composition after metabolizing cow's milk, and mainly include alcohols, methyl ketones and fatty acid ethyl esters. Example 2 (YJMK-3) produced a flavor compound with a total peak area of 2.3X10 7 About 2 times the total amount of flavor compounds produced in comparative example 1 (YJMK 1). Wherein the alcohols, ethyl esters and methyl ketones are about 1.9, 5.3 and 2.4 times that of comparative example 1, respectively. The alcohol content produced in example 2 is lower than that in comparative example 2, but the flavor substances contained in the alcohol-type milk are more abundant, and the ethyl ester compounds are more contained, so that the alcohol-type milk has fresh fruit and cream aroma and has great contribution potential to the flavor of the fermented milk.
Comparative example 3 (Lb1+S1) cow' S milk, when fermented, can detect 4 flavor compounds, 2-nonone, acetic acid, benzaldehyde and octanoic acid, respectively, with a total peak area of 0.7X10 7 .7 and 17 flavor compounds were detected in comparative example 3 (YJMK1+LB1+S10) and example 4 (YJMK3+LB1+S10), respectively, and the total peak areas were 1.6X10, respectively 7 And 2.7X10 7 . The alcohols, ethyl esters and methyl ketones of example 4 were 1.5, 7.6 and 2.3 times that of comparative example 3, respectively. As can be seen, the flavor compound composition of the cow milk co-fermented by YJMK-3 and lactic acid bacteria is richer and the content is higher than that of YJMK 1.
Table 3 comparative and example 2, 4 sensory evaluation tables
Sensory evaluation was performed in table 3 for comparative examples and examples 2 and 4, and the results are shown in fig. 5 and 6. Except for the degree of fineness, there was no significant difference between example 2 (YJMK-3) and comparative example 1 (YJMK 1), and the milk flavor, alcohol and taste of the fermented cow milk of example 2 were prominent as compared with comparative example 1. The wine taste of comparative example 2 is most obvious, but the milk flavor and taste of the fermented cow milk are covered up by the excessive wine taste.
Compared with comparative example 3 (LB 1+S10) and comparative example 4 (YJMK-3+LB1+S10), the milk flavor, taste and vinosity of example 4 (YJMK-3+LB1+S10) are more prominent, the taste is obvious, and the tissue state is good.
After the mother bacteria are subjected to expansion culture, 8 flavor compounds comprising 3 alcohols (ethanol, isoamyl alcohol and 2-heptanol), 2 methyl ketones (2-nonanone and 2-undecanone) and 3 ethyl esters (ethyl caproate, ethyl caprylate and ethyl caprate) can be independently fermented to obtain the cow milk, wherein the 3 major compounds account for 83.8%, 7.1% and 9.1%, and meanwhile, the cow milk has a brake taste. After being compounded with lactobacillus, YJMK3 is fermented to obtain cow milk, the obtained cow milk contains 14 flavor substances, and besides the flavor compounds, 4 kinds of acids (acetic acid, caproic acid, caprylic acid and capric acid), benzaldehyde and delta decalactone are produced, so that the cow milk has more abundant flavor substances and obvious stopping taste.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims (20)

1. Kluyveromyces marxianus (Kluyveromyces marxianus) YJMK-3 is characterized in that the Kluyveromyces marxianus is preserved in the China general microbiological culture Collection center (CGMCC) with the preservation number of 25655.
2. The kluyveromyces marxianus YJMK-3 of claim 1, wherein the medium of kluyveromyces marxianus YJMK-3 comprises the following components: glucose 35-45g/L, yeast extract 2.5-5.0g/L, soytone 10-15g/L.
3. The kluyveromyces marxianus YJMK-3 of claim 2, wherein the pH of the medium is adjusted to 6.5±0.2.
4. A kluyveromyces marxianus YJMK-3 as set forth in claim 2 or 3, wherein the conditions for the expanded culture of kluyveromyces marxianus YJMK-3 include: culturing for 10-16 h at 30+/-2 ℃.
5. The kluyveromyces marxianus YJMK-3 of claim 4, wherein the expanded culture conditions of kluyveromyces marxianus YJMK-3 include: culturing for 10-16 h at 30 ℃.
6. A microbial agent comprising the kluyveromyces marxianus YJMK-3 strain of any one of claims 1-5.
7. The microbial agent of claim 6, further comprising lactic acid bacteria;
the lactobacillus is one or more of Lactobacillus bulgaricus, streptococcus thermophilus, lactococcus lactis and Leuconostoc mesenteroides.
8. The microbial agent of claim 7, wherein the lactic acid bacteria are lactobacillus bulgaricus and streptococcus thermophilus.
9. The microbial agent according to claim 7 or 8, wherein the ratio of the number of viable bacteria of kluyveromyces marxianus YJMK-3 to the number of viable bacteria of the lactic acid bacteria is: 3: 1-1: 5.
10. the microbial agent according to claim 7 or 8, wherein the ratio of the number of viable bacteria of lactobacillus bulgaricus to the number of viable bacteria of streptococcus thermophilus is 1: 5-5: 1.
11. a fermentation product prepared from the kluyveromyces marxianus YJMK-3 of any one of claims 1-5 or the microbial inoculum of any one of claims 6-10 by fermentation culture.
12. The fermented product according to claim 11, characterized in that the fermented product is fermented milk, fermented whey or an aerated fermented beverage;
the gas-containing fermented beverage is prepared from fermented milk or fermented whey.
13. A method for producing a fermented product, wherein the fermented product is a fermented milk, comprising: adding Kluyveromyces marxianus YJMK-3 as defined in claim 1 into sterilized raw milk, or combining and inoculating Kluyveromyces marxianus YJMK-3 as defined in claim 1 with lactobacillus to obtain a sample to be fermented; the inoculation amount is 2% -6%;
fermenting the prepared sample at 30+/-5 ℃ for 12-36 hours, or fermenting at 40-42 ℃ for 4-6 hours, cooling to 30+/-5 ℃ and continuously fermenting for 12-24 hours to obtain fermented milk;
or, the fermented product is fermented whey, and the preparation method comprises the following steps:
adding YJKM-3 Kluyveromyces marxianus or combining, compounding and inoculating YJKM-3 Kluyveromyces marxianus and lactobacillus into sterilized raw whey to obtain a sample to be fermented; the inoculation amount is 2% -6%;
fermenting the prepared sample at 30+/-2 ℃ for 12-36 hours, or fermenting at 40-42 ℃ for 4-6 hours, cooling to 30+/-2 ℃ and continuing fermenting for 12-24 hours to obtain fermented whey;
or, the fermented product is an aerated fermented beverage, and the preparation method comprises the following steps: the fermented milk or fermented whey is used as a raw material, and one or more of water, a sour agent, a thickening emulsifier, a sweetener, fruit and vegetable juice and essence are added or not added, and the fermented milk or fermented whey is prepared by blending, homogenizing and sterilizing.
14. The method for preparing a fermented product according to claim 13, wherein the inoculation ratio of kluyveromyces marxianus YJMK-3 to lactic acid bacteria is 3: 1-1: 5.
15. the method for preparing a fermented product according to claim 14, wherein the inoculation ratio of kluyveromyces marxianus YJMK-3 to lactic acid bacteria is 1:1.
16. the method for producing a fermented product according to any one of claims 13 to 15, wherein the lactic acid bacteria include lactobacillus bulgaricus and one or more of streptococcus thermophilus, lactococcus lactis and leuconostoc mesenteroides.
17. A method of preparing a fermented product according to any one of claims 13-15, characterized in that when the fermented product is fermented milk or fermented whey, stirring is stopped during fermentation.
18. Use of kluyveromyces marxianus YJMK-3 as defined in any one of claims 1-5 or the microbial inoculum as defined in any one of claims 6-10 in any one of the following:
1) The taste of the fermented beverage is improved;
2) Adding flavor substances to the fermented product;
3) The mellow flavor of the fermented product is improved;
4) Improving the quality of fermented milk, fermented whey or gas-containing fermented beverage;
5) Promoting the metabolism of protein by lactic acid bacteria;
6) Improving the milk metabolism ability;
7) Provides a fermented beverage with unique flavor.
19. The use according to claim 18, wherein the flavour substances comprise one or more of alcohols, ketones, esters, acids.
20. The use according to claim 18 or 19, wherein the flavour material comprises one or more of ethanol, isoamyl alcohol, 2-heptanol, 2-nonanone, 2-undecanone, ethyl caproate, ethyl caprylate, ethyl caprate, acetic acid, caproic acid, caprylic acid, capric acid, benzaldehyde, and delta decalactone.
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