CN113462585A - Salt-tolerant yeast for increasing content of ethyl ester compounds in soy sauce and application thereof - Google Patents
Salt-tolerant yeast for increasing content of ethyl ester compounds in soy sauce and application thereof Download PDFInfo
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- CN113462585A CN113462585A CN202110593290.5A CN202110593290A CN113462585A CN 113462585 A CN113462585 A CN 113462585A CN 202110593290 A CN202110593290 A CN 202110593290A CN 113462585 A CN113462585 A CN 113462585A
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- 235000013555 soy sauce Nutrition 0.000 title claims abstract description 146
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 100
- 125000004494 ethyl ester group Chemical group 0.000 title claims abstract description 55
- 238000000855 fermentation Methods 0.000 claims abstract description 24
- 230000004151 fermentation Effects 0.000 claims abstract description 24
- 238000000034 method Methods 0.000 claims abstract description 11
- 238000004321 preservation Methods 0.000 claims abstract description 4
- 238000002360 preparation method Methods 0.000 claims abstract 3
- 238000009629 microbiological culture Methods 0.000 claims abstract 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 98
- 150000003839 salts Chemical class 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 26
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 24
- 239000011780 sodium chloride Substances 0.000 claims description 20
- 244000068988 Glycine max Species 0.000 claims description 19
- 235000010469 Glycine max Nutrition 0.000 claims description 19
- 235000013312 flour Nutrition 0.000 claims description 19
- 238000011081 inoculation Methods 0.000 claims description 19
- 238000002156 mixing Methods 0.000 claims description 18
- 238000010563 solid-state fermentation Methods 0.000 claims description 14
- 238000010411 cooking Methods 0.000 claims description 12
- 238000002791 soaking Methods 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 11
- 229940041514 candida albicans extract Drugs 0.000 claims description 10
- 239000012138 yeast extract Substances 0.000 claims description 10
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 8
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 8
- 240000006439 Aspergillus oryzae Species 0.000 claims description 6
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 6
- 238000010025 steaming Methods 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 238000011160 research Methods 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 2
- 230000002906 microbiologic effect Effects 0.000 claims 1
- 239000000796 flavoring agent Substances 0.000 abstract description 45
- 235000019634 flavors Nutrition 0.000 abstract description 45
- LZCLXQDLBQLTDK-UHFFFAOYSA-N ethyl 2-hydroxypropanoate Chemical compound CCOC(=O)C(C)O LZCLXQDLBQLTDK-UHFFFAOYSA-N 0.000 abstract description 16
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 abstract description 12
- -1 ester compounds Chemical class 0.000 abstract description 8
- 229940116333 ethyl lactate Drugs 0.000 abstract description 8
- MDHYEMXUFSJLGV-UHFFFAOYSA-N phenethyl acetate Chemical compound CC(=O)OCCC1=CC=CC=C1 MDHYEMXUFSJLGV-UHFFFAOYSA-N 0.000 abstract description 8
- JLIDRDJNLAWIKT-UHFFFAOYSA-N 1,2-dimethyl-3h-benzo[e]indole Chemical compound C1=CC=CC2=C(C(=C(C)N3)C)C3=CC=C21 JLIDRDJNLAWIKT-UHFFFAOYSA-N 0.000 abstract description 6
- DULCUDSUACXJJC-UHFFFAOYSA-N benzeneacetic acid ethyl ester Natural products CCOC(=O)CC1=CC=CC=C1 DULCUDSUACXJJC-UHFFFAOYSA-N 0.000 abstract description 6
- 230000015784 hyperosmotic salinity response Effects 0.000 abstract description 5
- 230000008569 process Effects 0.000 abstract description 2
- PWLNAUNEAKQYLH-UHFFFAOYSA-N butyric acid octyl ester Natural products CCCCCCCCOC(=O)CCC PWLNAUNEAKQYLH-UHFFFAOYSA-N 0.000 abstract 1
- 229940017219 methyl propionate Drugs 0.000 abstract 1
- UUIQMZJEGPQKFD-UHFFFAOYSA-N n-butyric acid methyl ester Natural products CCCC(=O)OC UUIQMZJEGPQKFD-UHFFFAOYSA-N 0.000 abstract 1
- AIILTVHCLAEMDA-UHFFFAOYSA-N thiopropionic acid S-methyl ester Natural products CCC(=O)SC AIILTVHCLAEMDA-UHFFFAOYSA-N 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 28
- 230000001953 sensory effect Effects 0.000 description 18
- 238000012258 culturing Methods 0.000 description 13
- 238000011156 evaluation Methods 0.000 description 13
- 239000000126 substance Substances 0.000 description 13
- 238000000605 extraction Methods 0.000 description 10
- YSNWHRKJEKWJNY-UHFFFAOYSA-N ethyl 3-(methylthio)propionate Chemical compound CCOC(=O)CCSC YSNWHRKJEKWJNY-UHFFFAOYSA-N 0.000 description 8
- 150000002333 glycines Chemical class 0.000 description 7
- 210000000582 semen Anatomy 0.000 description 7
- 230000008859 change Effects 0.000 description 6
- 235000009508 confectionery Nutrition 0.000 description 6
- 150000002148 esters Chemical class 0.000 description 6
- 230000012010 growth Effects 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- HCRBXQFHJMCTLF-ZCFIWIBFSA-N ethyl (2r)-2-methylbutanoate Chemical compound CCOC(=O)[C@H](C)CC HCRBXQFHJMCTLF-ZCFIWIBFSA-N 0.000 description 4
- 239000003205 fragrance Substances 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 238000005070 sampling Methods 0.000 description 4
- 235000015067 sauces Nutrition 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- WDAXFOBOLVPGLV-UHFFFAOYSA-N ethyl isobutyrate Chemical compound CCOC(=O)C(C)C WDAXFOBOLVPGLV-UHFFFAOYSA-N 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 108020004463 18S ribosomal RNA Proteins 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- QUMXDOLUJCHOAY-UHFFFAOYSA-N alpha-methylbenzyl acetate Natural products CC(=O)OC(C)C1=CC=CC=C1 QUMXDOLUJCHOAY-UHFFFAOYSA-N 0.000 description 2
- 235000013405 beer Nutrition 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000001813 ethyl (2R)-2-methylbutanoate Substances 0.000 description 2
- 229940090910 ethyl 2-methylbutyrate Drugs 0.000 description 2
- PPXUHEORWJQRHJ-UHFFFAOYSA-N ethyl isovalerate Chemical compound CCOC(=O)CC(C)C PPXUHEORWJQRHJ-UHFFFAOYSA-N 0.000 description 2
- FKRCODPIKNYEAC-UHFFFAOYSA-N ethyl propionate Chemical compound CCOC(=O)CC FKRCODPIKNYEAC-UHFFFAOYSA-N 0.000 description 2
- 238000001319 headspace solid-phase micro-extraction Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 241000228212 Aspergillus Species 0.000 description 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 244000285963 Kluyveromyces fragilis Species 0.000 description 1
- 235000014663 Kluyveromyces fragilis Nutrition 0.000 description 1
- 241000235033 Zygosaccharomyces rouxii Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
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- 238000013124 brewing process Methods 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
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- 239000003085 diluting agent Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 150000002241 furanones Chemical class 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
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- 229910052757 nitrogen Inorganic materials 0.000 description 1
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/50—Soya sauce
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
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- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Soy Sauces And Products Related Thereto (AREA)
Abstract
The invention discloses salt-tolerant yeast for increasing the content of ethyl ester compounds in soy sauce and application thereof. The strain has a preservation number of CGMCC No.21635, and is preserved in China general microbiological culture Collection center (CGMCC) at 1 month and 15 days 2021. The strain can be applied to the preparation of fermented soy sauce. The strain has good salt tolerance, and if the strain is added in the process of soy sauce mash fermentation, the content of ester compounds, particularly ethyl ester compounds in soy sauce can be remarkably improved, such as ethyl acetate, 2-ethyl methyl propionate, 3-ethyl methylthiopropionate, 2-phenylethyl acetate, ethyl lactate, 2-ethyl methyl butyrate and ethyl phenylacetate, so that the fruity and floral characteristics of the soy sauce are obviously improved, and the strain can be used for fermentation of high-salt dilute soy sauce and improving the flavor and quality of soy sauce products.
Description
Technical Field
The invention belongs to the technical field of food fermentation processing, and particularly relates to salt-tolerant yeast for increasing the content of ethyl ester compounds in soy sauce and application thereof.
Background
Soy sauce is one of traditional fermented foods and has become a necessary seasoning for Chinese households. The unique color, smell and taste of the health-care beer can not separate from the interaction of different microorganisms in the brewing process, and simultaneously, the health-care beer also contains rich nutrient components and has certain health-care effect. In the 20 s of the 19 th century, basic indexes such as ammonia-base nitrogen in soy sauce were emphasized, but as the living standard of the soy sauce was improved, consumers had higher requirements for color, flavor and taste of the soy sauce, and high-quality soy sauce with good flavor and taste was pursued, so that more and more researchers focused on flavor research of the soy sauce.
The soy sauce aroma compounds mainly comprise a plurality of substances such as acids, alcohols, aldehydes, esters, ketones, phenols, furanones and the like, wherein the ethyl ester compounds play a sweet, rich and soft substrate role in the soy sauce and endow sweet aroma and fruit aroma (von clouds, the research on the change rule and the formation mechanism of key aroma substances of high-salt dilute soy sauce [ D ] of southern China university of science 2015 ]. Wherein the compounds imparting fruity flavor include ethyl acetate, ethyl propionate, ethyl isobutyrate, ethyl lactate, ethyl 2-methylbutyrate, and ethyl isovalerate, which have a relatively low flavor threshold in the range of 0.1-10 μ g/kg; whereas compounds imparting floral notes mainly comprise phenylethyl acetate (floral and honey notes) and phenylethyl acetate (floral notes), with scent thresholds of 249.6 and 155.5 μ g/kg, respectively. The ethyl ester compounds have important flavor contribution effect in high-salt dilute soy sauce, are mainly closely related to yeast enzyme catalysis effect, and are slightly derived from chemical reaction of substances. The salt-tolerant ester-producing saccharomyces cerevisiae can be bred, so that the contents of ethyl acetate and ethyl lactate can be effectively increased, and the flavor of sauce mash juice can be improved. Therefore, the functional yeast for screening the high-yield ethyl ester compounds and applying the functional yeast to a soy sauce brewing system are strengthened, and the method has important significance for improving the content of the ethyl ester compounds in the soy sauce and improving the flavor quality of the soy sauce.
Currently, many researchers focus on the main salt-tolerant aroma-producing yeasts in soy sauce, such as zygosaccharomyces rouxii and candida, which have better salt tolerance and aroma-producing characteristics, and pay relatively less attention to other yeast strains having aroma-producing advantages in soy sauce. These strains are relatively low in soy sauce content, but have a certain contribution to specific aroma compounds in soy sauce. Therefore, the selection of the aroma-producing yeast should be diversified, the aroma-producing characteristics of the aroma-producing yeast are researched, the optimal inoculation time and the inoculation amount are explored, the flavor of the soy sauce product is coordinated, and the soy sauce product is rich in layers, so that the effects of improving the flavor of the soy sauce and improving the quality of the soy sauce are realized.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide the salt-tolerant yeast for improving the content of the ethyl ester compounds in the soy sauce and the application thereof.
According to the scheme provided by the invention, the content of ethyl ester compounds in the soy sauce is improved in a targeted manner by knowing the characteristics of salt-tolerant yeasts and selecting the appropriate addition amount of the strain, the addition time and the culture temperature, so that the flavor and the quality of the soy sauce at the present stage are improved.
The purpose of the invention is realized by at least one of the following technical solutions.
The salt tolerant yeast SWJS W2009L 3 WJW for improving the content of the ethyl ester compounds in the soy sauce is a variable Vickers pseudostellaris (Wickerhamiellaverersatislis); it has the following morphological physiological characteristics: the colony is round, the diameter is 3mm, the colony is brown-white, the surface is wet, smooth and raised, and the microscopic examination shape is small round; the salt tolerance range (by sodium chloride NaCl tolerance concentration) is 0-20%, the optimum growth temperature is 30 ℃, and the optimum growth pH is 5.5.
The salt-tolerant yeast for improving the content of the ethyl ester compounds in the soy sauce is salt-tolerant variable pseudo-yeast (Wickerhamiellaverastis), is named as SWJS W2009L 3 WJW, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms in 1 month and 15 days 2021, and is addressed to the institute of microbiology of Zhongkou institute No. 3 of Zhonglu-xi-Lu-1 of the sunward area in Beijing, and the preservation number is CGMCC No. 21635.
The salt-tolerant yeast for improving the content of the ethyl ester compounds in the soy sauce can be applied to fermented soy sauce.
The invention provides an application of salt-tolerant yeast for improving the content of ethyl ester compounds in soy sauce in fermented soy sauce, which comprises the following steps:
(1) soaking and cooking: soaking soybean in water, heating for steaming, filtering to obtain residue, and collecting cooked soybean;
(2) mixing and yeast stirring: uniformly mixing the cooked beans, flour and yeast extract in the step (1) to obtain yeast;
(3) culturing the yeast for making hard liquor: carrying out solid state fermentation on the yeast in the step (2) to obtain finished yeast;
(4) fermentation of the soy sauce mash and inoculation of target bacteria: uniformly mixing the finished koji obtained in the step (3) with saline water to obtain soy sauce mash; and (2) carrying out primary fermentation treatment on the sauce mash, then inoculating the salt-tolerant yeast SWJS W2009L 3 WJW (salt-tolerant variable pseudo-vWicker yeast) for improving the content of ethyl ester compounds in the sauce, carrying out secondary fermentation treatment, sterilizing, filtering and taking filtrate to obtain the finished sauce.
Further, the soybeans in the step (1) are soaked in water for 6-10 h.
Further, the cooking treatment time in the step (1) is 10-15 min.
Further, the pressure of the cooking treatment in the step (1) is 0.1-0.12 MPa.
Further, the temperature of the cooking treatment in the step (1) is 121-125 ℃.
Further, the temperature of the cooked beans in the step (2) is 32-45 ℃.
Further, the mass ratio of the flour in the step (2) to the soybeans in the step (1) is 1:3-1: 4.
Further, the aspergillus in the step (2) is aspergillus oryzae with good activity.
Further, the inoculation amount of the koji in the step (2) is 0.04-0.5 wt% of the total mass of the soybeans and the flour.
Preferably, the mass ratio of the flour in the step (2) to the soybeans in the step (1) is 1: 4.
further, the solid state fermentation in step (3) comprises: fermenting at 28-30 deg.C for 16-24h, and fermenting at 26-28 deg.C for 16-26 h;
further, the first yeast turning is completed within 14h-18h after the solid state fermentation in the step (3) is started, and the second yeast turning is completed within 4h-7h after the first yeast turning is completed.
Further, the mass percentage concentration of the saline water in the step (4) is 18-22%;
further, the volume ratio of the saline water in the step (4) to the Daqu in the step (3) is 1:1.8-1: 2.5;
further, the time of the first fermentation treatment in the step (4) is 15-60 d;
further, the inoculation amount of the salt tolerant yeast for increasing the content of the ethyl ester compounds in the soy sauce in the step (4) is 104-105CFU/mL；
Further, the time of the second fermentation treatment in the step (4) is 30-75 d;
further, the first fermentation treatment and the second fermentation treatment in the step (4) are performed at the optimum temperature of the sun-dried night dew at room temperature.
According to the fermented soy sauce provided by the invention, the flavor substances can be measured by analyzing the volatile substance condition in the soy sauce by adopting a headspace solid phase microextraction method, and the ethyl ester compounds in the soy sauce are mainly analyzed; the sensory evaluation can be carried out by quantitative sensory analysis (QDA) to determine flavor change of soy sauce.
Further, the headspace solid-phase microextraction method comprises the steps of putting 4mL of fermentation liquor (the finished soy sauce) into a headspace sample injection bottle, adjusting the salt concentration to be 180g/L, and completing aroma extraction work by using an extraction head.
The invention screens out salt-tolerant yeast (a variable Vickers pseudovinum) from soy sauce mash, researches the influence of the salt-tolerant yeast on the flavor components of high-salt dilute soy sauce, and has certain innovation. And the method is applied to the soy sauce brewing production process, and has certain potential and value for improving the flavor of the soy sauce and improving the quality of the soy sauce.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the salt-tolerant yeast SWJS W2009L 3 WJW (Wickerhamiellaverastis) for improving the content of the ethyl ester compounds in the soy sauce, which is provided by the invention, is applied to soy sauce fermentation, can obviously improve the content of the ester compounds in the soy sauce, particularly the content of the ethyl ester compounds, such as ethyl acetate, ethyl 2-methylpropionate, ethyl 3-methylthiopropionate, 2-phenylethyl acetate, ethyl lactate, ethyl 2-methylbutyrate and ethyl phenylacetate, obviously improve the fruity and floral characteristics of the soy sauce, and the prepared soy sauce has more harmonious and lasting flavor.
Drawings
FIG. 1 is a plate morphology diagram of salt tolerant yeast strain SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in soy sauce in the example;
FIG. 2 is a microscope morphology diagram of a salt tolerant yeast strain SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in soy sauce in the example;
FIG. 3 is a growth curve of the salt tolerant yeast strain SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in soy sauce in the example;
fig. 4 is a salt tolerance detection result diagram of the salt tolerant yeast strain SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in soy sauce in the example.
Detailed Description
The following description of the embodiments of the present invention is provided in connection with the accompanying drawings and examples, but the invention is not limited thereto. It is noted that the processes described below, if not specifically described in detail, are all realizable or understandable by those skilled in the art with reference to the prior art. The reagents or apparatus used are not indicated to the manufacturer, and are considered to be conventional products available by commercial purchase.
1. Isolation and screening of strains
Under the aseptic condition, 2.5g of mashed high-salt dilute soy sauce mash sample is placed in an aseptic sealed bag with 25mL of aseptic normal saline to be uniformly mixed, 0.1mL of soy sauce mash sample diluent is placed in a PDA flat plate to be uniformly coated, the PDA flat plate is placed in an incubator at 28 ℃ to be cultured, and the salt-tolerant yeast for improving the content of ethyl ester compounds in the soy sauce is obtained by separation and purification and is named as SWJS W2009L 3 WJW.
2. Identification of strains
2.1 morphological physiological characteristics of the Strain
The colony is round, has a diameter of 3mm, is brownish white, has a wet, smooth and raised surface, and has a plate shape as shown in FIG. 1. The cell morphology under a microscope is shown in FIG. 2. The strain can grow when the NaCl concentration is 0-20%, and the salt tolerance is shown in figure 3; the optimum growth pH is 5.5; the optimal growth temperature is 30 ℃; the strain reached logarithmic growth phase at 55h, and the growth curve is shown in FIG. 4.
2.2 molecular biological identification
And (3) carrying out 18S rDNA sequencing on the separated salt-tolerant yeast SWJS W2009L 3 WJW for improving the content of the ethyl ester compounds in the soy sauce, and comparing the sequencing result to determine that the strain is variable Vickers pseudostellariella (Wickerhamiellaevis), wherein the 18S rDNA sequence of the strain is shown in SEQ ID No. 1. The salt-tolerant yeast SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in the soy sauce has the preservation number of CGMCC No.21635, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms at 1 month and 15 days in 2021, and is addressed to the microorganism research institute of Zhongkoyao No. 3 Hospital of West Lu No.1 Hospital in the morning area of Beijing.
Example 1
An application of salt-tolerant yeast for increasing the content of ethyl ester compounds in soy sauce in preparing fermented soy sauce comprises the following steps:
(1) soaking and cooking: adding semen glycines into water, soaking for 6 hr, and steaming at 0.1MPa for 15min at 121 deg.C under high temperature and high pressure, cooling to 32 deg.C, filtering to obtain residue, and collecting cooked semen glycines;
(2) mixing and yeast stirring: uniformly mixing the cooked beans, flour and yeast extract in the step (1), wherein the mass ratio of the flour to the soybeans in the step (1) is 1:4, aspergillus oryzae is used as yeast extract, and the inoculation amount is 0.04 wt% of the total mass of the soybeans and the flour, so that the yeast is obtained;
(3) culturing the yeast for making hard liquor: carrying out solid state fermentation on the Daqu obtained in the step (2), firstly culturing for 16h at 30 ℃, and then culturing for 28h at 28 ℃; wherein, the first yeast turning is completed within 15h after the solid state fermentation is started, and the second yeast turning is completed within 7h after the first yeast turning is completed, so as to obtain finished yeast;
(4) fermentation of the soy sauce mash and inoculation of target bacteria: uniformly mixing the finished koji in the step (3) with saline water, wherein the mass percentage concentration of the saline water is 18 wt%, and the volume ratio of the Daqu to the saline water in the step (3) is 1:2.5 to obtain soy sauce mash; fermenting the mash to 15d according to 104Inoculating the salt-tolerant yeast SWJS W2009L 3 WJW with the increased ethyl ester compound content in the soy sauce by the CFU/mL inoculation amount, continuing fermenting for 90d, and sterilizing and filtering to obtain filtrate to obtain the finished soy sauce.
(5) Determination of flavor substances: putting 4mL of fermentation liquor (the finished soy sauce) into a headspace sampling bottle, adjusting the salt concentration to 180g/L, and finishing the fragrance extraction work by using an extraction head.
(6) Sensory evaluation: the flavor change of the finished soy sauce was determined by sensory evaluation panel (male 3, female 5) and was performed by quantitative sensory descriptive method (QDA).
Comparative example 1: comparative example 1 is substantially the same as example 1, except that the salt tolerant yeast SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in the soy sauce is not added in step (4), and the rest conditions are the same, to obtain a soy sauce. The soy sauce of comparative example 1 was also subjected to the tests of the above-mentioned step (5) and step (6).
Analysis of results
Compared with the comparative example 1, the total amount of the ethyl ester compounds in the finished soy sauce is remarkably increased to 2.44 times of that in the comparative example 1 after the salt tolerant yeast SWJS W2009L 3 WJW for increasing the content of the ethyl ester compounds in the soy sauce is inoculated in the example 1, wherein the contents of ethyl lactate (fruity flavor) and ethyl phenylacetate (flowery flavor) are respectively 3.77 times and 5.43 times of that in the comparative example 1, and meanwhile, ester substances such as ethyl 3-methylthiopropionate (fruity flavor and sweet flavor) can be newly generated, and the table 1 can be referred.
Also, the sensory evaluation scores of the fruity and floral notes of the finished soy sauce prepared in example 1 were 2.38 times and 2.60 times, respectively, as compared with those of comparative example 1, as shown in table 2.
Example 2
An application of salt-tolerant yeast for increasing the content of ethyl ester compounds in soy sauce in preparing fermented soy sauce comprises the following steps:
(1) soaking and cooking: adding semen glycines into water, soaking for 7 hr, and steaming at 123 deg.C under 0.1MPa for 13min at high temperature and high pressure, cooling to 35 deg.C, filtering to obtain residue, and collecting cooked semen glycines;
(2) mixing and yeast stirring: uniformly mixing the cooked beans, flour and yeast extract in the step (1), wherein the mass ratio of the flour to the soybeans in the step (1) is 1:4, aspergillus oryzae is used as yeast extract, and the inoculation amount is 0.2 wt% of the total mass of the soybeans and the flour, so that the yeast is obtained;
(3) culturing the yeast for making hard liquor: performing solid state fermentation on the Daqu obtained in the step (2), culturing for 22h at 30 ℃, and then culturing for 22h at 27 ℃; wherein, the first yeast turning is finished at 16h after the solid state fermentation is started, and the second yeast turning is finished after 6h after the first yeast turning is finished;
(4) fermentation of the soy sauce mash and inoculation of target bacteria: uniformly mixing the finished koji in the step (3) with saline water, wherein the mass percentage concentration of the saline water is 19 wt%, and the volume ratio of the Daqu to the saline water in the step (3) is 1:2.2 to obtain soy sauce mash; fermenting the mash to 30d according to 104Inoculating the salt-tolerant yeast SWJS W2009L 3 WJW with the increased ethyl ester compound content in the soy sauce by the CFU/mL inoculation amount, continuing fermenting for 90d, and sterilizing and filtering to obtain filtrate to obtain the finished soy sauce.
(5) Determination of flavor substances: putting 4mL of fermentation liquor (the finished soy sauce) into a headspace sampling bottle, adjusting the salt concentration to 180g/L, and finishing the fragrance extraction work by using an extraction head.
(6) Sensory evaluation: the flavor change of the finished soy sauce was determined by sensory evaluation panel (male 3, female 5) and was performed by quantitative sensory descriptive method (QDA).
Comparative example 2: comparative example 2 is substantially the same as example 2, except that the salt tolerant yeast SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in the soy sauce is not added in step (4), and the rest conditions are the same, so that a soy sauce is obtained. The soy sauce of comparative example 2 was also subjected to the tests of the above-mentioned step (5) and step (6).
Analysis of results
Compared with the comparative example 2, the total amount of the ethyl ester compounds in the finished soy sauce is remarkably increased to 2.10 times of that of the comparative example 2 after the salt tolerant yeast SWJS W2009L 3 WJW for increasing the content of the ethyl ester compounds in the soy sauce is inoculated in the example 2, wherein the content of ethyl lactate (fruity flavor) ethyl phenylacetate (flower flavor) is 3.46 times and 5.30 times of that of the comparative example 2 respectively, and meanwhile ester substances such as ethyl 3-methylthiopropionate (fruity flavor and sweet flavor) can be newly generated, which is shown in table 1.
Also, the sensory evaluation scores of the fruity flavor and the floral flavor of the finished soy sauce prepared in example 2 were 1.75 times and 2.00 times, respectively, as compared with those of comparative example 2, as shown in table 2.
Example 3
An application of salt-tolerant yeast for increasing the content of ethyl ester compounds in soy sauce in preparing fermented soy sauce comprises the following steps:
(1) soaking and cooking: adding semen glycines into water, soaking for 10 hr, steaming at 125 deg.C under 0.12MPa for 15min, cooling to 45 deg.C, and filtering to obtain residue.
(2) Mixing and yeast stirring: uniformly mixing the cooked beans, flour and yeast extract in the step (1), wherein the mass ratio of the flour to the soybeans in the step (1) is 1:4, aspergillus oryzae is used as yeast extract, and the inoculation amount is 0.5 wt% of the total mass of the soybeans and the flour, so that the yeast is obtained;
(3) culturing the yeast for making hard liquor: performing solid state fermentation on the Daqu obtained in the step (2), firstly culturing for 24h at 30 ℃, and then culturing for 20h at 28 ℃; wherein, the first yeast turning is completed within 16h after the solid state fermentation is started, and the second yeast turning is completed within 7h after the first yeast turning is completed, so as to obtain finished yeast;
(4) fermentation of the soy sauce mash and inoculation of target bacteria: uniformly mixing the finished koji in the step (3) with saline water, wherein the mass percentage concentration of the saline water is 20 wt%, and the volume ratio of the Daqu to the saline water in the step (3) is 1:2.3 to obtain soy sauce mash; fermenting the mash to 30d according to 105Inoculating the salt-tolerant yeast SWJS W2009L 3 WJW with the increased ethyl ester compound content in the soy sauce by the CFU/mL inoculation amount, continuing fermenting for 90d, and sterilizing and filtering to obtain filtrate to obtain the finished soy sauce.
(5) Determination of flavor substances: putting 4mL of fermentation liquor (the finished soy sauce) into a headspace sampling bottle, adjusting the salt concentration to 180g/L, and finishing the fragrance extraction work by using an extraction head.
(6) Sensory evaluation: the flavor change of the finished soy sauce was determined by sensory evaluation panel (male 3, female 5) and was performed by quantitative sensory descriptive method (QDA).
Comparative example 3: comparative example 3 is substantially the same as example 3, except that the salt tolerant yeast SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in the soy sauce is not added in step (4), and the rest conditions are the same, to obtain a soy sauce. The soy sauce of comparative example 3 was also subjected to the tests of the above-mentioned step (5) and step (6).
Analysis of results
Compared with the comparative example 3, in the example 3, after the salt tolerant yeast SWJS W2009L 3 WJW for increasing the content of the ethyl ester compounds in the soy sauce is inoculated, the total amount of the ethyl ester compounds in the finished soy sauce is remarkably increased to be 2.92 times of that of the comparative example 3, wherein the content of ethyl lactate (fruity flavor) and the content of ethyl phenylacetate (flowery flavor) are respectively 5.08 times and 7.42 times of that of the comparative example 3, meanwhile, ester substances such as 3-methylthioethyl propionate (fruity flavor and sweet flavor) can be newly generated, the fruity flavor and flowery flavor characteristics of the soy sauce are remarkably improved, and the table 1 shows.
Also, the sensory evaluation scores of the fruity and floral notes of the finished soy sauce prepared in example 3 were 3.33 times and 3.38 times, respectively, as compared with those of comparative example 3, as shown in Table 2.
Example 4
An application of salt-tolerant yeast for increasing the content of ethyl ester compounds in soy sauce in preparing fermented soy sauce comprises the following steps:
(1) soaking and cooking: adding semen glycines into water, soaking for 8 hr, and steaming at high temperature and high pressure under 0.12MPa for 14min at 124 deg.C, and cooling to 38 deg.C to obtain cooked semen glycines;
(2) mixing and yeast stirring: uniformly mixing the cooked beans in the step (1) with flour and yeast extract, wherein the mass ratio of the flour to the soybeans in the step (1) is 1:3, aspergillus oryzae is used as yeast extract, and the inoculation amount is 0.15 wt% of the total mass of the soybeans and the flour, so that the yeast is obtained;
(3) culturing the yeast for making hard liquor: performing solid state fermentation on the Daqu obtained in the step (2), firstly culturing for 24h at 30 ℃, and then culturing for 20h at 28 ℃; wherein, the primary yeast turning is completed at 18h after the solid state fermentation is started, and the secondary yeast turning is completed after 8h to obtain finished yeast;
(4) fermentation of the soy sauce mash and inoculation of target bacteria: uniformly mixing the finished koji in the step (3) with saline water, wherein the mass percentage concentration of the saline water is 22 wt%, and the volume ratio of the Daqu to the saline water in the step (3) is 1:2.5 to obtain soy sauce mash; fermenting the mash to 60d according to 104Inoculating the salt-tolerant yeast SWJS W2009L 3 WJW with the increased ethyl ester compound content in the soy sauce by the CFU/mL inoculation amount, continuing fermenting for 90d, and sterilizing and filtering to obtain filtrate to obtain the finished soy sauce.
(5) Determination of flavor substances: putting 4mL of fermentation liquor (the finished soy sauce) into a headspace sampling bottle, adjusting the salt concentration to 180g/L, and finishing the fragrance extraction work by using an extraction head.
(6) Sensory evaluation: the flavor change of the finished soy sauce was determined by sensory evaluation panel (male 3, female 5) and was performed by quantitative sensory descriptive method (QDA).
Comparative example 4: comparative example 4 is substantially the same as example 4, except that the salt tolerant yeast SWJS W2009L 3 WJW for increasing the content of ethyl ester compounds in the soy sauce is not added in step (4), and the rest conditions are the same, to obtain a soy sauce. The soy sauce of comparative example 4 was also subjected to the tests of the above-mentioned step (5) and step (6).
Analysis of results
Compared with the comparative example 4, in the example 4, after the salt tolerant yeast SWJS W2009L 3 WJW for increasing the content of the ethyl ester compounds in the soy sauce is inoculated, the total amount of the ethyl ester compounds in the finished soy sauce is remarkably increased to be 1.93 times of that of the comparative example 4, wherein the content of ethyl lactate (fruity flavor) and the content of ethyl phenylacetate (flowery flavor) are respectively 2.26 times and 4.17 times of that of the comparative example 4, and meanwhile, ester substances such as ethyl 3-methylthiopropionate (fruity flavor and sweet flavor) can be newly generated, which can be shown in table 1.
Also, the sensory evaluation scores of the fruity and floral notes of the finished soy sauce prepared in example 4 were 1.43 times and 1.75 times, respectively, as compared with those of comparative example 4, as shown in Table 2.
TABLE 1
Note: in Table 1, it indicates that no detection was made
TABLE 2
The above examples are only preferred embodiments of the present invention, which are intended to be illustrative and not limiting, and those skilled in the art should understand that they can make various changes, substitutions and alterations without departing from the spirit and scope of the invention.
Sequence listing
<110> university of southern China's science
GUANGZHOU INSTITUTE OF MODERN INDUSTRIAL TECHNOLOGY
<120> salt-tolerant yeast for increasing content of ethyl ester compounds in soy sauce and application thereof
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<213> salt tolerant versatile yeast pseudowilcoxiellavia strains (Wickerhamiellaverastis)
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tgctgacgct atgcagagtg tcgaaaagaa tggagatgta cacgttagtg aaacttgtac 360
accaagcttt ctttaaaaga cctcgtatca ggcaagacta cccgctgaac ttaagcatat 420
caataagcg 429
Claims (10)
1. The salt-tolerant yeast for increasing the content of ethyl ester compounds in soy sauce is characterized by being named as SWJS W2009L 3 WJW, being preserved in China general microbiological culture Collection center at 1 month and 15 days of 2021, being the microbiological research institute of Zhongkou institute of Navy No.1 Hospital No. 3 of North West Lu province in the Chaoyang district of Beijing, and having the preservation number of CGMCC No. 21635.
2. The use of the salt tolerant yeast according to claim 1 for increasing the content of ethyl esters in soy sauce in the preparation of fermented soy sauce.
3. The application of the salt tolerant yeast for increasing the content of ethyl ester compounds in soy sauce according to claim 2 in preparing fermented soy sauce is characterized by comprising the following steps:
(1) soaking and cooking: soaking soybean in water, heating for steaming, filtering to obtain residue, and collecting cooked soybean;
(2) mixing and yeast stirring: uniformly mixing the cooked beans, flour and yeast extract in the step (1) to obtain yeast;
(3) carrying out solid state fermentation on the yeast in the step (2) to obtain finished yeast;
(4) uniformly mixing the finished koji obtained in the step (3) with saline water to obtain soy sauce mash; and (3) performing primary fermentation treatment on the soy sauce mash, then inoculating the salt-tolerant yeast for improving the content of ethyl ester compounds in the soy sauce, performing secondary fermentation treatment, sterilizing, filtering and taking filtrate to obtain the finished soy sauce.
4. The application of the salt tolerant yeast for increasing the content of ethyl ester compounds in soy sauce in preparing fermented soy sauce according to claim 3, wherein the soybeans in the step (1) are soaked in water for 6-10 h; the time of the cooking treatment in the step (1) is 10-15min, the pressure of the cooking treatment is 0.1-0.12MPa, and the temperature of the cooking treatment is 121-125 ℃.
5. The use of the salt tolerant yeast for increasing the content of ethyl esters in soy sauce according to claim 3, wherein the temperature of the cooked beans in step (2) is 32-45 ℃.
6. The application of the salt tolerant yeast for increasing the content of ethyl ester compounds in soy sauce in preparing fermented soy sauce according to claim 3, wherein the mass ratio of the flour in the step (2) to the soybeans in the step (1) is 1:3-1: 4; the koji in the step (2) is aspergillus oryzae, and the inoculation amount of the koji is 0.04-0.5 wt% of the total mass of the soybeans and the flour.
7. The use of the halotolerant yeast for increasing the content of ethyl esters in soy sauce according to claim 3 in the preparation of fermented soy sauce, wherein the solid-state fermentation in step (3) comprises: fermenting at 28-30 deg.C for 16-24h, and fermenting at 26-28 deg.C for 16-26 h; and finishing the first yeast turning within 14-18h after the solid state fermentation is started, and finishing the second yeast turning within 4-7h after the first yeast turning is finished.
8. The application of the salt tolerant yeast for increasing the content of ethyl ester compounds in soy sauce in preparing fermented soy sauce according to claim 3, wherein the salt water in the step (4) has a mass percentage concentration of 18-22%; the volume ratio of the saline water in the step (4) to the Daqu in the step (3) is 1:1.8-1: 2.5.
9. The use of the halotolerant yeast for increasing the content of ethyl esters in soy sauce according to claim 3, wherein the time of the first fermentation treatment in step (4) is 15-60 days; the inoculation amount of the salt tolerant yeast for increasing the content of the ethyl ester compounds in the soy sauce is 104-105CFU/mL; the time of the second fermentation treatment in the step (4) is 30-75 days.
10. A fermented soy sauce produced by the method of use according to any one of claims 3 to 9.
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