CN110115366A - A method of improving Cantonese high-salt diluted state fermentation soy fragrance - Google Patents
A method of improving Cantonese high-salt diluted state fermentation soy fragrance Download PDFInfo
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- CN110115366A CN110115366A CN201910295577.2A CN201910295577A CN110115366A CN 110115366 A CN110115366 A CN 110115366A CN 201910295577 A CN201910295577 A CN 201910295577A CN 110115366 A CN110115366 A CN 110115366A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 51
- 230000004151 fermentation Effects 0.000 title claims abstract description 51
- 238000000034 method Methods 0.000 title claims abstract description 38
- 239000003205 fragrance Substances 0.000 title claims abstract description 30
- 239000001963 growth medium Substances 0.000 claims abstract description 63
- 150000003839 salts Chemical class 0.000 claims abstract description 51
- 235000013555 soy sauce Nutrition 0.000 claims abstract description 36
- 238000004519 manufacturing process Methods 0.000 claims abstract description 15
- 238000005057 refrigeration Methods 0.000 claims abstract description 7
- 238000009423 ventilation Methods 0.000 claims abstract description 5
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical group CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 60
- 241000894006 Bacteria Species 0.000 claims description 39
- 239000004310 lactic acid Substances 0.000 claims description 30
- 235000014655 lactic acid Nutrition 0.000 claims description 30
- 241000894007 species Species 0.000 claims description 30
- 241000235342 Saccharomycetes Species 0.000 claims description 28
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 25
- 239000008103 glucose Substances 0.000 claims description 25
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 239000002054 inoculum Substances 0.000 claims description 8
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 7
- 239000002253 acid Substances 0.000 claims description 7
- 238000010899 nucleation Methods 0.000 claims description 7
- 235000005979 Citrus limon Nutrition 0.000 claims description 6
- 244000131522 Citrus pyriformis Species 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- 235000015278 beef Nutrition 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 239000010779 crude oil Substances 0.000 claims description 6
- 150000004985 diamines Chemical class 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 238000011534 incubation Methods 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 229940099596 manganese sulfate Drugs 0.000 claims description 6
- 239000011702 manganese sulphate Substances 0.000 claims description 6
- 235000007079 manganese sulphate Nutrition 0.000 claims description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 6
- 229920000053 polysorbate 80 Polymers 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 239000001632 sodium acetate Substances 0.000 claims description 6
- 235000017281 sodium acetate Nutrition 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 239000000654 additive Substances 0.000 claims description 5
- 230000000996 additive effect Effects 0.000 claims description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 230000003519 ventilatory effect Effects 0.000 claims description 4
- 230000000694 effects Effects 0.000 abstract description 20
- 235000015067 sauces Nutrition 0.000 abstract description 9
- 239000000243 solution Substances 0.000 description 35
- 230000007423 decrease Effects 0.000 description 7
- 238000001514 detection method Methods 0.000 description 5
- 235000019441 ethanol Nutrition 0.000 description 5
- 239000012531 culture fluid Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 150000001298 alcohols Chemical class 0.000 description 3
- 230000007812 deficiency Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- NBBJYMSMWIIQGU-UHFFFAOYSA-N Propionic aldehyde Chemical compound CCC=O NBBJYMSMWIIQGU-UHFFFAOYSA-N 0.000 description 2
- 150000001299 aldehydes Chemical class 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000004099 anaerobic respiration Effects 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 150000002241 furanones Chemical class 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229940063896 nitrogen 60 % Drugs 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 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
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
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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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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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
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- 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
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- Biochemistry (AREA)
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- Soy Sauces And Products Related Thereto (AREA)
Abstract
The invention discloses a kind of methods for improving Cantonese high-salt diluted state fermentation soy fragrance, it is cultivated the following steps are included: the salt tolerant aroma former parent species of refrigeration are forwarded in parent species triangular flask culture medium, is then switched to culture in production triangular flask culture medium and obtains salt tolerant aroma former original seed；Will through S1, treated that salt tolerant aroma former original seed is forwarded in seed tank culture base cultivates, then by strain transfer into fermentation tank culture medium, cultivate so that culture solution viable count reaches 108More than；The pH for adjusting and keep S2 treated culture solution, stands, ventilation is kept during standing；S4: it will be added in the soy sauce to ferment by the strain in S3 treated culture solution.This method may insure the salt tolerant aroma former spawn activity with higher added during sauce fermentation, to effectively improve the fragrance of obtained soy sauce.
Description
Technical field
The invention belongs to soy sauce brewing fields, and in particular to a kind of side for improving Cantonese high-salt diluted state fermentation soy fragrance
Method.
Background technique
Soy sauce fragrance is derived mainly from the volatile component in soy sauce, the types of these volatile components up to hundreds of,
Following a few classes can be substantially divided into: 1. alcohols: referring mainly to methanol, ethyl alcohol, propyl alcohol, butanol etc., can generate esters with organic acid reaction；
2. aldehydes: mainly acetaldehyde, propionic aldehyde, vanillic aldehyde etc. are aoxidized during sauce fermentation by corresponding alcohol, and it is special to have
Fragrance；3. organic acid: being formed during the fermentation by alcohols, Oxidation of aldehydes, there is unique fragranced；4. esters: being to have
Machine acid is esterified with corresponding alcohols, has unique aromatic odor；5. phenols: the phenolic precursors substance mainly in wheat
It acts on and generating through saccharomycete；6. Furanones: being that saccharomycete is metabolized during the fermentation, it has soft saline taste, and can rise
To the effect for increasing taste effect.
Cantonese high-salt fermentation technique is improved on China's traditional zymotic Process ba- sis, while drawing Japan again
Many advanced sauce fermentation technologies, fermentation period are 3~6 months, and soy sauce amino-acid nitrogen production rate is high, accounts for about full nitrogen
60% or so.
Compared with Japanese soy sauce, the disadvantage of the generally existing fragrance deficiency of most domestic soy sauce: traditional Cantonese is with high salt dilute
State zymotechnique is the zymotechnique of weather exposure under natural conditions, and no added producing fragrant strains, rely primarily on shine it is natural in tank
Phase effect generates soy sauce fragrance to bacterial strain in boundary after fermentation, and the fermented product of this zymotechnique, quality fluctuation is greatly and by season
Section factor influences, it is possible to soy sauce made of finally fermenting be caused to there is a problem of fragrance deficiency.Therefore, in order to improve Cantonese height
The stability of salt lean state zymotechnique needs on the basis of the technique, and salt tolerant aroma former is added during sauce fermentation, with
Increase the fragrance of soy sauce.
Before salt tolerant aroma former is added to the soy sauce of fermentation, need first to expand culture it.Traditional salt tolerant produces fragrant
Bacterium expands culture process: test tube strains (parent species) are transferred triangular flask kind (original seed), and original seed is forwarded to kind with 10% inoculum concentration
Sub- tank is then directly used among sauce fermentation by seeding tank with 10% inoculum concentration switching fermentation tank culture.It is this traditional
Salt tolerant aroma former expands culture and combines China's Technology of Brewing Soy Sauce feature, and salt tolerant aroma former often can not be after expanding culture
It is added immediately in the soy sauce to ferment.Its spawn activity will appear the feelings of decline during the storage of salt tolerant aroma former
Condition.And Various Seasonal salt tolerant aroma former is different by the degree of environmental influence, so that salt tolerant aroma former spawn activity changes
The case where it is different.It is added in the case where spawn activity is low in the soy sauce of fermentation, the effect of salt tolerant aroma former can be made significantly
It reduces, Cantonese soy sauce obtained is caused the problem of fragrance deficiency occur.
Summary of the invention
The purpose of the present invention is to provide a kind of methods for improving Cantonese high-salt diluted state fermentation soy fragrance, and this method can be with
The salt tolerant aroma former spawn activity with higher for ensuring to add during sauce fermentation, to effectively improve obtained sauce
The fragrance of oil.
The purpose of the present invention is achieved through the following technical solutions:
A method of improving Cantonese high-salt diluted state fermentation soy fragrance, comprising the following steps:
S1: the salt tolerant aroma former parent species after refrigeration culture being forwarded in parent species triangular flask culture medium and are cultivated, then will be female
Culture solution and parent species in kind triangular flask are switched to culture in multiple production triangular flask culture mediums and obtain salt tolerant aroma former
Original seed；
S2: will through S1, treated that salt tolerant aroma former original seed is forwarded in seed tank culture base cultivates, then by strain transfer
Into fermentation tank culture medium, under conditions of 33~35 DEG C, cultivate 2~5 days, so that culture solution viable count reaches 108More than；
S3: being adjusted and keeps S2 treated the pH of culture solution in the range of 6~8, is stood under conditions of 0~10 DEG C
10~60 days, ventilation is kept during standing；
S4: it will be added in the soy sauce to ferment by the strain in S3 treated culture solution.
The principle of the present invention is:
The growth of strain is divided into four-stage, is germination period, growth period, logarithmic phase and decline phase respectively, in logarithmic phase,
Nutriment is sufficient, and the quantity appearance of strain is significantly grown.In the process, it is influenced by environmental factors such as weathers,
Strain is easily accessible decline phase, and strain mortality causes salt tolerant aroma former overall activity to decline.If directly using in the case
In sauce fermentation, since the vigor of strain is insufficient, cause the fragrance of soy sauce made of fermentation insufficient.
The present invention and the prior art the difference is that, after being cultivated in strain and fermentation tank culture medium, first stand
One specific time, then participate in the fermentation of soy sauce.It is proved after inventor's long-term experiment, in condition of the present invention
The lower salt tolerant aroma former for having stood special time period, spawn activity is relatively high during entire strain is survived, and can guarantee hair
Soy sauce fragrance after ferment is significantly improved.
As one embodiment of the present invention, the salt tolerant aroma former is saccharomycete M24.
When the salt tolerant aroma former be saccharomycete M24 when, the present invention preferably:
The time that S3 is stood is 60 days, and glucose, the addition of the glucose were added into culture solution at interval of 10 days
Amount is 3%, and the percentage is mass percent, on the basis of the gross mass of culture solution.
As another embodiment of the invention, the salt tolerant aroma former is lactic acid bacteria M30.
When the salt tolerant aroma former be lactic acid bacteria M30 when, the present invention preferably:
The time that S3 is stood is 15 days；PH value is preferably 6.5~7.5.
Currently preferred static conditions are all that the best item for specific strain is being confirmed after experiment
Part, after standing with this condition, saccharomycete M24 and lactic acid bacteria M30 can respectively reach higher bacterial activity, subsequent experimental
Also confirm that the soy sauce fragrance made of the participation fermentation of these bacterium is stronger than the soy sauce of the prior art.
Unlike saccharomycete M24, lactic acid bacteria M30 will do it anaerobic respiration during growth and generate lactic acid,
PH value is caused to decline.Lactic acid build to a certain extent after, can lactic acid bacteria inhibiting M30 growth.Therefore, for lactic acid bacteria M30
For, the pH control during standing is more stringent.
Parent species triangular flask culture medium of the present invention, the recommendation of production triangular flask culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
9.5~10.5 parts of peptone
9.5~10.5 parts of beef extract powder
4.5~5.5 parts of yeast extract
18~22 parts of glucose
1.5~2.5 parts of lemon acid diamine
1.5~2.5 parts of dipotassium hydrogen phosphate
4.5~5.5 parts of sodium acetate
0.04~0.06 part of manganese sulfate
0.15~0.25 part of magnesium sulfate
0.5~1.5 part of Tween 80
80~120 parts of salt；
(2) pH to 6.8~7.2 of culture medium is adjusted with 20~40% sodium hydroxide solution, and at 121~125 DEG C
Under the conditions of handle 30~40min, be subsequently cooled to 30~35 DEG C.
The saccharomycete seeding tank culture medium and yeast fermentation tank culture medium is prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
200~250 parts of the soy sauce crude oil of fermentation
18~22 parts of glucose
80~120 parts of salt；
(2) 30min is handled under conditions of 125 DEG C, is subsequently cooled to 30 DEG C.
It, will be through S1 treated salt tolerant aroma former original seed during the S2 as a kind of preferred embodiment of the invention
It is forwarded in seed tank culture base with 5~10% inoculum concentration, under conditions of 33~35 DEG C, is cultivated 2~3 days, incubation
Middle holding ventilatory capacity and culture volume ratio are 1:1~1:1.5.
Compared with the prior art, the invention has the following beneficial effects:
The method of the invention stands the specific time by the salt tolerant aroma former after it will cultivate, to obtain spawn activity
The spawn activity of high salt tolerant aroma former culture solution, this method gained culture solution is stablized, and Cantonese high-salt dilute is applied to
In fermented sauce, the production perfume (or spice) effect of salt tolerant aroma former itself can be effectively played, the resulting soy sauce that ferments gives off a strong fragrance, quality
It is high.
Specific embodiment
Below by way of specific embodiment, the present invention is further illustrated.
Embodiment is all made of method of dilution butteron on plate detection living cells quantity.For soy sauce brewing, in order to enable after fermentation
Soy sauce fragrance can occur obviously being promoted, the viable count of salt tolerant aroma former at least needs to reach 108More than.
Embodiment 1
The influence of time of repose and temperature to saccharomycete M24 spawn activity
S1: parent species triangular flask culture medium, production triangular flask culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
9.5 parts of peptone；10.5 parts of beef extract powder；5 parts of yeast extract；18 parts of glucose；1.5 parts of lemon acid diamine；
1.5 parts of dipotassium hydrogen phosphate；4.5 parts of sodium acetate；0.06 part of manganese sulfate；0.15 part of magnesium sulfate；1.5 parts of Tween 80；80 parts of salt；
(2) pH to 7.0 of culture medium is adjusted with 20% sodium hydroxide solution, and is handled under conditions of 121 DEG C
30min is subsequently cooled to 30 DEG C；
S2: saccharomycete seeding tank culture medium and yeast fermentation tank culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
200 parts of the soy sauce crude oil of fermentation；18 parts of glucose；80 parts of salt；
(2) 30min is handled under conditions of 125 DEG C, is subsequently cooled to 30 DEG C；
S3: the saccharomycete M24 parent species after refrigeration culture are forwarded in parent species triangular flask culture medium made from S1 and are cultivated, so
Afterwards by parent species triangular flask culture solution and saccharomycete M24 parent species be switched to multiple productions and cultivated in triangular flask culture medium
Obtain saccharomycete M24 original seed；
S4: by through S3, treated that saccharomycete M24 original seed is forwarded in seed tank culture base with 5% inoculum concentration cultivates,
It under conditions of 35 DEG C, cultivates 2 days, ventilation is kept in incubation, keep ventilatory capacity and culture volume ratio is 1:1, then will
Strain transfer is into fermentation tank culture medium, under conditions of 35 DEG C, cultivates 3 days, so that bacterium cell number reaches 108More than；
S5: by S4, treated that saccharomycete M24 stands 60 under conditions of 0~10 DEG C, 10~20 DEG C, 20~30 DEG C respectively
It carries out one-time detection every 10 days cell numbers to strain, and experimental result is as shown in following table -1:
The viable count of -1 saccharomycete M24 culture solution of table standing different temperatures and time
| Time of repose | 0~10 DEG C | 10~20 DEG C | 20~30 DEG C |
| 10d | 5.2×108 | 7.1×108 | 4.2×108 |
| 20d | 2.1×108 | 2.6×108 | 1.2×108 |
| 30d | 4.2×108 | 7.8×107 | 4.6×107 |
| 40d | 5.2×108 | 4.5×107 | 2.2×107 |
| 50d | 4.8×108 | 2.6×107 | 1.2×107 |
| 60d | 7.5×108 | 1.3×107 | 2.6×106 |
By table -1 as it can be seen that saccharomycete M24 culture solution is stored in respectively in 0~10 DEG C, 10~20 DEG C, 20~30 DEG C of condition
Lower to stand 60 days, the viable count in yeast bacteria culture fluid constantly declines with the extension of time of repose；But 0~10 DEG C storage
The decline of culture solution spawn activity it is relatively slow, upon standing between when reaching 60 days, viable count is 7.5 × 108, it is able to satisfy soy sauce brewing
10 are reached for salt tolerant aroma former viable count8Requirement.Therefore, 0~10 DEG C of M24 yeast bacteria culture fluid storage condition selection, it is quiet
The culture solution for setting 60 days saccharomycete M24 can ensure that the fragrance of soy sauce is significantly improved.
Embodiment 2
Influence of the glucose additive amount to saccharomycete M24 spawn activity
S1: parent species triangular flask culture medium, production triangular flask culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
10.5 parts of peptone；9.5 parts of beef extract powder；4.5 parts of yeast extract；22 parts of glucose；Lemon acid diamine 2.5
Part；2.5 parts of dipotassium hydrogen phosphate；5.5 parts of sodium acetate；0.04 part of manganese sulfate；0.25 part of magnesium sulfate；0.5 part of Tween 80；Salt 120
Part；
(2) 40min is handled under conditions of 125 DEG C, is subsequently cooled to 35 DEG C；
S2: saccharomycete seeding tank culture medium and yeast fermentation tank culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
250 parts of the soy sauce crude oil of fermentation；22 parts of glucose；120 parts of salt；
40min is handled under conditions of 121 DEG C, is subsequently cooled to 35 DEG C；
S3: the saccharomycete M24 parent species after refrigeration culture are forwarded in parent species triangular flask culture medium made from S1 and are cultivated, so
Afterwards by parent species triangular flask culture solution and saccharomycete M24 parent species be switched to multiple productions and cultivated in triangular flask culture medium
Obtain saccharomycete M24 original seed；
S4: by through S3, treated that saccharomycete M24 original seed is forwarded in seed tank culture base with 10% inoculum concentration cultivates,
It under conditions of 33 DEG C, cultivates 2 days, ventilation is kept in incubation, keep ventilatory capacity and culture volume ratio is 1:1.5, then
By strain transfer into fermentation tank culture medium, under conditions of 35 DEG C, cultivate 3 days, so that bacterium cell number reaches 108More than；
S5: by S4, treated that saccharomycete M24 stands 60 days under the conditions of 0~10 DEG C, and every 10d adds a glucose,
Glucose additive amount is respectively 0%, 1%, 2%, 3%, 4%, carries out one-time detection, experiment every 10 days cell numbers to strain
As a result as shown in following table -2:
The saccharomycete M24 culture solution that table -2 adds different amounts of glucose stands the viable count of different time
| Time of repose | 0% | 1% | 2% | 3% | 4% |
| 10d | 5.2×108 | 5.5×109 | 6.0×109 | 7.5×108 | 7.3×109 |
| 20d | 2.1×108 | 2.6×109 | 3.2×109 | 6.3×108 | 4.4×109 |
| 30d | 4.2×108 | 1.3×109 | 1.4×109 | 5.2×108 | 2.3×109 |
| 40d | 5.2×108 | 7.8×108 | 8.0×108 | 3.6×109 | 1.0×109 |
| 50d | 4.8×108 | 5.2×108 | 6.0×108 | 6.9×108 | 7.5×108 |
| 60d | 7.5×108 | 3.6×108 | 6.7×108 | 1.2×109 | 6.9×108 |
By table -2 as it can be seen that every 10d adds a glucose, then when M24 yeast bacteria culture fluid dwell temperature is 0~10 DEG C
Vigor increases compared to the case where not adding glucose during saccharomycete storage, when glucose additive amount is 3%, ferment
Vigor highest during female bacterium storage, is to compare with the culture solution viable count for not adding glucose during storage.Therefore, ferment
When female bacteria culture fluid is stored under the conditions of 0~10 DEG C, every 10d adds the glucose of 3% additive amount, so that during storage
Culture solution vigor improve.
Embodiment 3
The influence of time of repose and temperature to lactic acid bacteria M30 spawn activity
S1: parent species triangular flask culture medium, production triangular flask culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
10 parts of peptone；10 parts of beef extract powder；5.5 parts of yeast extract；20 parts of glucose；2 parts of lemon acid diamine；Phosphoric acid
2 parts of hydrogen dipotassium；5 parts of sodium acetate；0.05 part of manganese sulfate；0.2 part of magnesium sulfate；1 part of Tween 80；100 parts of salt；
(2) pH to 7 of culture medium is adjusted with 30% sodium hydroxide solution, and handles 35min under conditions of 122 DEG C,
It is subsequently cooled to 32 DEG C；
S2: lactic acid bacteria seeding tank culture medium and lactobacillus-fermented tank culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
220 parts of the soy sauce crude oil of fermentation；20 parts of glucose；100 parts of salt；
(2) pH to 6.8 of culture medium is adjusted with 25% sodium hydroxide solution, and is handled under conditions of 122 DEG C
40min is subsequently cooled to 32 DEG C.
S3: the lactic acid bacteria M30 parent species after refrigeration culture are forwarded in parent species triangular flask culture medium made from S1 and are cultivated, so
Afterwards by parent species triangular flask culture solution and lactic acid bacteria M30 parent species be switched to multiple productions and cultivated in triangular flask culture medium
Obtain lactic acid bacteria M30 original seed；
S4: by through S3, treated that lactic acid bacteria M30 original seed is forwarded in seed tank culture base with 7% inoculum concentration cultivates,
Under conditions of 34 DEG C, cultivate 5 days, static in incubation, closed culture, then by strain transfer into fermentation tank culture medium,
Under conditions of 35 DEG C, cultivate 6 days, so that bacterium cell number reaches 108More than；
S5: by S4, treated that lactic acid bacteria M30 stands 20 under conditions of 0~10 DEG C, 10~20 DEG C, 20~30 DEG C respectively
It carries out one-time detection every 5 days cell numbers to strain, and experimental result is as shown in following table -3:
The viable count of -3 lactic acid bacteria M30 culture solution of table standing different temperatures and time
| Time of repose | 0~10 DEG C | 10~20 DEG C | 20~30 DEG C |
| 5d | 4.6×108 | 7.3×108 | 3.7×108 |
| 10d | 2.1×108 | 2.7×108 | 1.3×108 |
| 15d | 6.4×108 | 7.3×107 | 3.7×107 |
| 20d | 5.2×108 | 4.3×107 | 2.2×107 |
By table -3 as it can be seen that lactic acid bacteria M30 culture solution is stored in respectively in 0~10 DEG C, 10~20 DEG C, 20~30 DEG C of condition
Lower to stand 20 days, the viable count in lactic acid bacteria culture solution constantly declines with the extension of time of repose；But 0~10 DEG C storage
The decline of culture solution spawn activity it is relatively slow, upon standing between when reaching 15 days, viable count reaches highest 6.4 × 108, Er Qieneng
Meet soy sauce brewing and 10 are reached for salt tolerant aroma former viable count8Requirement.Therefore, M30 lactic acid bacteria culture solution storage condition selects
With 0~10 DEG C, standing 15 days lactic acid bacteria culture solutions can ensure that the fragrance of soy sauce is significantly improved.
Embodiment 4
Influence of the pH value to lactic acid bacteria M30 spawn activity
S1: parent species triangular flask culture medium, production triangular flask culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
10.5 parts of peptone；10 parts of beef extract powder；5 parts of yeast extract；19 parts of glucose；1.5 parts of lemon acid diamine；Phosphorus
2.5 parts of sour hydrogen dipotassium；4.5 parts of sodium acetate；0.05 part of manganese sulfate；0.2 part of magnesium sulfate；1 part of Tween 80；90 parts of salt；
(2) pH to 7.2 of culture medium is adjusted with 20% sodium hydroxide solution, and is handled under conditions of 123 DEG C
30min is subsequently cooled to 30 DEG C.
The lactic acid bacteria seeding tank culture medium and lactobacillus-fermented tank culture medium is prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
210 parts of the soy sauce crude oil of fermentation；20 parts of glucose；90 parts of salt；
(2) pH to 6.5 of culture medium is adjusted with 20% sodium hydroxide solution, and is handled under conditions of 123 DEG C
30min is subsequently cooled to 35 DEG C.
S3: the lactic acid bacteria M30 parent species after refrigeration culture are forwarded in parent species triangular flask culture medium made from S1 and are cultivated, so
Afterwards by parent species triangular flask culture solution and lactic acid bacteria M30 parent species be switched to multiple productions and cultivated in triangular flask culture medium
Obtain lactic acid bacteria M30 original seed；
S4: by through S3, treated that lactic acid bacteria M30 original seed is forwarded in seed tank culture base with 8% inoculum concentration cultivates,
Under conditions of 33 DEG C, cultivates 5 days, remain stationary in incubation, anaerobic state, then by strain transfer to fermentation tank culture medium
In, under conditions of 35 DEG C, cultivate 5 days, so that bacterium cell number reaches 108More than；
S5: S4 treated lactic acid bacteria M30 is stood 20 under conditions of pH is 6.0,6.5,7.0,7.5 and 8.0 respectively
It, temperature is maintained between 0~10 DEG C, carries out one-time detection, experimental result such as following table -4 every 5 days viable counts to culture solution
It is shown:
- 4 lactic acid bacteria M30 culture solution of table stands the viable count of different pH value and time
| Time of repose | pH 6.0 | pH 6.5 | pH 7.0 | pH 7.5 | pH 8.0 |
| 5d | 4.6×108 | 5.0×109 | 5.8×109 | 5.2×109 | 4.3×109 |
| 10d | 2.1×108 | 2.6×109 | 3.2×109 | 2.8×109 | 1.2×109 |
| 15d | 6.4×108 | 1.2×109 | 2.2×109 | 1.0×109 | 8.7×108 |
| 20d | 5.2×108 | 7.8×108 | 1.5×109 | 7.3×108 | 6.2×108 |
By the data of table -4 as it can be seen that when pH value is between 6.5~7.5 when standing 15 days, lactic acid bacteria M30's
Viable count reaches maximum, and is able to satisfy soy sauce brewing and reaches 10 for salt tolerant aroma former viable count8Requirement.
It should be pointed out that above-described embodiment be only to further explanation of the invention, rather than limit, art technology
Any adjustment or change of the personnel in the comparable meaning and scope with technical solution of the present invention are all considered as being included in this
In the protection scope of invention.
Claims (8)
1. a kind of method for improving Cantonese high-salt diluted state fermentation soy fragrance, which comprises the following steps:
S1: the salt tolerant aroma former parent species of refrigeration being forwarded in parent species triangular flask culture medium and are cultivated, then will be in parent species triangular flask
Culture solution and parent species be switched in multiple production triangular flask culture mediums culture and obtain salt tolerant aroma former original seed；
S2: will through S1, treated that salt tolerant aroma former original seed is forwarded in seed tank culture base cultivates, then by strain transfer to sending out
In fermentation tank culture medium, under conditions of 33~35 DEG C, cultivate 2~5 days, so that culture solution viable count reaches 108More than；
S3: being adjusted and keeps S2 treated the pH of culture solution in the range of 6~8, stand 10 under conditions of 0~10 DEG C~
60 days, ventilation is kept during standing；
S4: it will be added in the soy sauce to ferment by the strain in S3 treated culture solution.
2. the method according to claim 1 for improving Cantonese high-salt diluted state fermentation soy fragrance, which is characterized in that described
Salt tolerant aroma former is saccharomycete M24.
3. the method according to claim 2 for improving Cantonese high-salt diluted state fermentation soy fragrance, which is characterized in that S3 is stood
Time be 60 days, and at interval of glucose is added within 10 days into culture solution, the additive amount of the glucose is 3%, described hundred
Divide than being mass percent, on the basis of the gross mass of culture solution.
4. the method according to claim 1 for improving Cantonese high-salt diluted state fermentation soy fragrance, which is characterized in that described
Salt tolerant aroma former is lactic acid bacteria M30.
5. the method according to claim 4 for improving Cantonese high-salt diluted state fermentation soy fragrance, which is characterized in that S3 is stood
Time be 15 days；PH value is preferably 6.5~7.5.
6. described in any item methods for improving Cantonese high-salt diluted state fermentation soy fragrance, feature exist according to claim 1~5
In the parent species triangular flask culture medium, the recommendation of production triangular flask culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
9.5~10.5 parts of peptone
9.5~10.5 parts of beef extract powder
4.5~5.5 parts of yeast extract
18~22 parts of glucose
1.5~2.5 parts of lemon acid diamine
1.5~2.5 parts of dipotassium hydrogen phosphate
4.5~5.5 parts of sodium acetate
0.04~0.06 part of manganese sulfate
0.15~0.25 part of magnesium sulfate
0.5~1.5 part of Tween 80
80~120 parts of salt；
(2) pH to 6.8~7.2 of culture medium is adjusted with 20~40% sodium hydroxide solution, and under conditions of 121~125 DEG C
30~40min is handled, is subsequently cooled to 30~35 DEG C.
7. the method according to claim 6 for improving Cantonese high-salt diluted state fermentation soy fragrance, which is characterized in that described
Saccharomycete seeding tank culture medium and yeast fermentation tank culture medium are prepared by the following method:
(1) culture medium is prepared according to following components and proportion:
200~250 parts of the soy sauce crude oil of fermentation
18~22 parts of glucose
80~120 parts of salt；
(2) 30min is handled under conditions of 125 DEG C, is subsequently cooled to 30 DEG C.
8. the method according to claim 6 for improving Cantonese high-salt diluted state fermentation soy fragrance, which is characterized in that the S2
It will be forwarded in seed tank culture base through S1 treated salt tolerant aroma former original seed with 5~10% inoculum concentration in the process, 33~
It under conditions of 35 DEG C, cultivates 2~3 days, ventilatory capacity is kept in incubation and culture volume ratio is 1:1~1:1.5.
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