CN107488591A - A kind of high enzyme activity, high salt tolerant soy sauce bacterial strain selection and its application process - Google Patents
A kind of high enzyme activity, high salt tolerant soy sauce bacterial strain selection and its application process Download PDFInfo
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Abstract
The present invention relates to a kind of high enzyme activity, the aspergillus oryzae strain of high salt tolerant, selection and its application process in high saline diluting Technology of Brewing Soy Sauce, aspergillus oryzae strain deposit number is CCTCC NO:M2017077.The selection of the present invention is for starting strain with aspergillus oryzae 3.951, is passed through60Co gamma-ray irradiations and chemical reagent joint mutagenesis, high salt coating primary dcreening operation culture, preferably culture obtain high enzyme activity, the aspergillus oryzae strain of high salt tolerant.The resistance to high salt of bacterial strain of seed selection of the present invention, koji spore count are more, germination percentage is high, enzyme activity is high, heritability is stable, can effectively improve protein utilization, starch sugar rate, and then improve soy sauce oil yield and reach the purpose for making full use of brewing materials；And the soy sauce flavor components content prepared using bacterial strain of the present invention is significantly improved, and has typical paste flavor and distinctive flavor component, so that flavor of soy sauce is more abundant, pure and sweet.
Description
Technical field
It is resistance to more particularly, to a kind of high enzyme activity, height the present invention relates to the technical field of microorganism fungus kind selection
Aspergillus oryzae strain, selection and its application process of salt.
Background technology
Soy sauce is one of with a long history, standby ace-high traditional condiment in China.China's soy sauce annual production is up to more than 500 ten thousand
Ton, accounts for world's annual production more than 60%.Sauce fermentation mainly has two methods of low-salt solid method and high saline diluting.At present I
State's soy sauce is mainly based on low-salt solid method, but the soy sauce quality of method production is not as good as high-salt fermentation method.As people give birth to
Running water is flat to be improved, and the demand of high-quality soy sauce is also continuously increased.The soy sauce quality formed height, wind are brewed by high saline diluting
Taste is good, and the salt of high concentration can not only play anti-corrosion and antibacterial effect, moreover it is possible to assign soy sauce saline taste and delicate flavour.But equipment for making sauce
The metabolic activity and caused protease of crucial strain aspergillus oryzae used in oil, the enzyme activity of amylase and its stability also can be by
Reduced to the influence of high salt factor, so as to cause the speed of decomposing protein and starch materials and utilization rate to reduce, amino
Acid-state nitrogen yield rate is low, and starch sugar rate is low.Meanwhile yeast fermenting carbohydrate generates alcohol, lactobacillus-fermented in fermentation process
The change of soy sauce color, flavour that a series of carbohydrate generation biochemical reactions such as lactic acid and Maillard reaction are brought slows down or subtracted
Less, the problems such as fermenting-ripening is slower, and fermentation period is long is also following.Japanese soy sauce is contained with its free sugar, total nitrogen and organic acid
The advantages that amount is higher occupies the market share of a large amount of overseas markets and high-grade soy sauce, although its production method and China's tradition
Production method it is similar, but the protein utilization of its raw material is ranked first in the world with more than 90% height, and China's tradition high salt is dilute
Protein utilization only has 75%-80% in state soy sauce brewing, this microorganism fungus kind seed selection advanced with Japan, composite bacteria
The technology such as koji-making and multiple bacteria compound fermentation is inseparable.Therefore, by induced mutations breeding high-yield quality strains, so as to obtain
Hyperosmosis can be resistant in hypersaline environment by obtaining, and high proteinase yield, amylase and flavor substance are abundant, quality is high, heredity
The stable aspergillus oryzae strain of shape is significant.
The content of the invention
The present invention provides a kind of high enzyme activity, the aspergillus oryzae strain of high salt tolerant, selection and its application process, for gram
The technical problem that high saline diluting brews high-quality soy sauce can not be applicable by taking existing microorganism fungus kind.
3rd, to achieve the above object, the present invention provides a kind of high enzyme activity, the aspergillus oryzae strain of high salt tolerant, and deposit number is
CCTCC NO:M2017077。
A kind of described high enzyme activity, the form of aspergillus oryzae (Aspergillus oryzae) bacterial strain of high salt tolerant are as follows：Bacterium
Fall rounded, a diameter of 35~40mm, quality is loose, and mycelia is slightly shorter compared with starting strain, but grows vigorous, and color is yellow green,
The back side is colourless.Conidial head is radial, 200~300 μm of diameter, 2~3mm of sporophore, the μ of diameter 10~20 at nearly top capsule
M, wall is thin and coarse, and top capsule is spherical, about 25 μm.
The technical solution adopted by the present invention is：
A kind of high enzyme activity, high salt tolerant aspergillus oryzae strain selection, comprise the following steps：(1) starting strain is connect
Kind is selected the flat board to grow fine and carried out in cultivating 60~90h on casein agar plating medium60Co- gamma-ray irradiations lure
Become, Induced dosage chooses 1~5kGy, puts after having radiated and is separated into desinfection chamber, continues 60~90h of culture, according to mycelia
Growing state and bacterium colony transparent circle size, eugonic bacterial strain is filtered out by control group of starting strain.
(2) by the inoculation that step (1) screens on PDA slant mediums, in 25~30 DEG C of cultures to sprouting
After phase, with normal saline flushing slant medium, thalline is collected by centrifugation, adds physiological saline and obtains spore count as 106~107Individual/
Ml spore suspension；
(3) take mutagens stoste to add and mutagens mother liquor is configured into buffer solution, hanged in the spore described in step (1)
With volume ratio 1 in supernatant liquid：3~5 add mutagens mother liquor, are centrifuged after handling 60~120min at 25~30 DEG C, with buffering
Liquid rinses to reaction terminating repeatedly；
(4) spore suspension after step (3) processing is respectively coated and put down in the casein agar for adding sodium chloride solution
Cultivated on plate culture medium, according to mycelial growth situation and bacterium colony transparent circle size, growth is filtered out by control group of starting strain
Vigorous bacterium colony, and be forwarded to immediately on wort agar inclined-plane culture medium, obtain primary dcreening operation bacterium to ripe in 25~30 DEG C of cultures
Strain, it is standby to be subsequently placed in 4 DEG C of refrigerations；
(5) preferred culture medium is prepared by a certain percentage with analysis for soybean powder, wheat-middlings, wheat bran, water, the preferred culture that will be prepared
Base is dispensed to triangular flask, 121 DEG C of 25~30min of sterilizing, the primary dcreening operation inoculation described in the ring step (4) of picking 1 to triangular flask
It is interior, mix after 25~30 DEG C of accumulation 60~90h of culture, determine its spore count, germination percentage and enzymatic activity, spore count highest, hair
Bud rate and enzyme activity highest bacterial strain are high enzyme activity, the aspergillus oryzae strain of high salt tolerant.
Preferably, the starting strain described in step (1) is aspergillus oryzae 3.951.
Preferably, the mutagens described in step (3) are ethylmethane sulfonates, and described buffer solution is that pH is 7.0~8.0
Phosphate buffer.
Preferably, the initial mass fraction of the sodium chloride solution described in step (4) is 10~14%, and incremental gradient is
2%, final mass fraction is 22~26%.
Preferably, step (1), the recipe ingredient of casein agar plating medium described in (4) are 15 parts by weight
Pancreatic digest of casein, 5 parts of soy meal papain digestion things, 5 parts of sodium chloride, 15 parts of agar, 1000 parts of distilled water；
The recipe ingredient of described wort agar inclined-plane culture medium by weight for 130 parts of fructus hordei germinatus leaching powders, 0.1 part of chloramphenicol, 5 parts
Sodium chloride, 15 parts of agar, 1000 parts of distilled water.
Preferably, the recipe ingredient of the preferred culture medium described in step (5) be by weight 7.2 parts of analysis for soybean powder, 2 parts times
Powder, 0.8 part of wheat bran, 30 parts of distilled water.
A kind of high enzyme activity, the aspergillus oryzae strain of high salt tolerant are applied to high-salt diluted state fermentation soy brewage process.
A kind of high enzyme activity, the application process of aspergillus oryzae strain of high salt tolerant are：
A, 7.2 portions of soybean, 2 parts of wheat-middlings, 0.8 part of wheat bran are weighed in parts by weight, after soybean is soaked into 3~5 hours
Water is put only, takes out soya bean, adds the wheat bran weighed up, loads boiling in normal pressure steamer, to steam to well-done without rotten, bean cotyledon
It is suitable to be split up into, then is mixed with wheat-middlings, spreads out and carries out being cooled to 25~30 DEG C, accesses high enzyme activity, the aspergillus oryzae of high salt tolerant
Bacterial strain, inoculum concentration 0.2%, mix, in 30 DEG C of accumulation culture 72h, then song is made into.
B, by the Cheng Quyu salt solution of making according to 1：2~5 mass ratio is added in sauce fermentation technique, enters one
Preferable mass ratio is walked as 1：3.
The guarantor that biological material specimens of the present invention have been specified according to the regulation of Patent Law detailed rules for the implementation in State Intellectual Property Office
Hide unit --- China typical culture collection center preservation, address：Chinese Wuhan Wuhan Universitys, preservation date 2017 3
The moon 1；Deposit number is CCTCC NO:M2017077；Classification And Nomenclature：Aspergillus oryzae HNCC 6.189, Latin name：
Asporgillus orgzoe HNCC6.189；Biological specimen viability is detected on March 11st, 2017 and completed, and result is to deposit
It is living.
The beneficial effects of the present invention are：
1st, the present invention uses induced mutation breeding method, passes through60Co- gamma-ray irradiations and chemical reagent ethylmethane sulfonate joint
Mutagenesis, carries out induced mutations breeding high-yield quality strains to the bacterial strain of aspergillus oryzae 3.951, successful incubation is a kind of have resistance to high salt,
Koji spore count is more, germination percentage is high, enzyme activity is high, heritability is stable and is easily enlarged the rice of numerous good characteristics such as culture
Aspergillus strain (Aspergillus oryzae) HNCC 6.189.
2nd, aspergillus oryzae strain (Aspergillus oryzae) HNCC 6.189 of seed selection of the present invention is by the egg of starting strain
White matter utilization rate brings up to 89% from 80%, improves positive effect, then by improving protein utilization, starch sugar rate, enters
And improve soy sauce oil yield and reach the purpose for making full use of brewing materials；Secondly, the aspergillus oryzae strain of seed selection of the present invention
The high-salt dilute soy production cycle was shortened to 5 by (Aspergillus oryzae) HNCC 6.189 from original 6 months
Month, large-scale industrial production is adapted to, it has a extensive future.
3rd, soy sauce is made with setting out in aspergillus oryzae (Aspergillus oryzae) bacterial strain HNCC 6.189 of seed selection of the present invention
Soy sauce is compared made from strain fermentation, and the content of its amino-acid nitrogen, total acid and flavor substance is all significantly larger than starting strain
In terms of the content of soy sauce made from fermentation, wherein flavor substance alcohols material relative amount add 1.17% citronellol and
0.63% ethanol, aldehydes matter add 0.19% 4-ethyl guaiacol, and acid adds the 2,4- of 0.25%
Adipic acid, 1.11% caproic acid and 1.57% syringic acid, Ester add 3.75% ethyl acetate, 0.06%
Methyl stearate, aldoketones material add 1.53% 5- methyl -2- furfurals, 5.66%4- hydroxyl -2- ethyl -5- methyl -
3 (2H)-furanones, 1.09% 2,5- dimethyl-pyrazins and 0.23% 2- furfurals, above flavor substance mostly have allusion quotation
The paste flavor of type and distinctive flavor component, make flavor of soy sauce more abundant, pure and sweet.
Embodiment
Below in conjunction with the table 1-4 in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out it is clear,
It is fully described by, it is clear that described embodiment is only part of the embodiment of the present invention, rather than whole embodiments.Base
Embodiment in the present invention, ordinary skill people obtained under the premise of creative work is not made it is all its
His embodiment, belongs to the scope of protection of the invention.
Embodiment 1：
A kind of high enzyme activity, the aspergillus oryzae strain of high salt tolerant, depositary institution are China typical culture collection center preservation,
Address：Chinese Wuhan Wuhan Universitys, preservation date on March 1st, 2017；Deposit number is CCTCC NO:M2017077；Point
Class is named：Aspergillus oryzae HNCC 6.189, Latin name：Asporgillus orgzoeHNCC 6.189；Biological specimen is survived
Property on March 11st, 2017 detect complete, result be survival.
A kind of high enzyme activity, high salt tolerant aspergillus oryzae strain selection, comprise the following steps：
(1) starting strain is inoculated on casein agar plating medium and cultivates 72h, select the flat board to grow fine,
Send and carried out in Hunan Province's nuclear agricultural science and space breeding research institute60Co- gamma-ray irradiation mutagenesis, choose 1kGy, 2kGy, 3kGy and make
For Induced dosage, each 10 flat boards of dose delivery, totally 30 flat boards., will be through with black cloth after having radiated60Co- gamma-ray irradiations
Flat board after mutagenic treatment, which is wrapped, to be brought back, and is put and is separated into desinfection chamber, continues to cultivate 72h, according to mycelial growth situation
With bacterium colony transparent circle size, eugonic bacterial strain is filtered out by control group of starting strain.
(2) by the inoculation that step (1) screens on PDA slant mediums, in 25~30 DEG C of cultures to sprouting
After phase, with normal saline flushing slant medium, thalline is collected by centrifugation, adds physiological saline and obtains spore count as 106~107Individual/
Ml spore suspension；
(3) take ethylmethane sulfonate stoste to add into the phosphate buffer that pH is 7.2 and be configured to ethylmethane sulfonate mother
Liquid, 8ml ethylmethane sulfonate mother liquor is added in 2ml described spore suspension, after 25~30 DEG C handle 90min
Centrifuge, rinsed repeatedly to reaction terminating with phosphate buffer；
(4) by step (3) processing after spore suspension be respectively coated sodium chloride solution mass fraction be 12%,
14%th, cultivated on 16%, 18%, 20%, 22%, 24% casein agar plating medium, and strain be numbered,
Numbering is respectively Lp001, Lp002, Lp003, Lp004, Lp005, Lp006, Lp007 (wherein, casein agar flat board culture
The recipe ingredient of base is by weight 15 parts of pancreatic digest of casein, 5 parts of soy meal papain digestion things, 5 parts of chlorine
Change sodium, 15 parts of agar, 1000 parts of distilled water)；Then according to mycelial growth situation and bacterium colony transparent circle size, with starting strain
Aspergillus oryzae 3.951 is that control group filters out eugonic bacterium colony, and is forwarded to immediately on wort agar inclined-plane culture medium,
After ripening in 4 days is cultivated in 25~30 DEG C and obtains primary dcreening operation bacterial strain, and it is standby to be subsequently placed in 4 DEG C of refrigerations.(wherein, wort agar inclined-plane
The recipe ingredient of culture medium is by weight 130 parts of fructus hordei germinatus leaching powders, 0.1 part of chloramphenicol, 5 parts of sodium chloride, 5.0,15 parts of agar
15.0th, 1000 parts of distilled water)
(5) 7.2 parts of analysis for soybean powder, 2 parts of wheat-middlings, 0.8 part of wheat bran, 30 parts of distilled water are weighed by weight prepares preferred culture
Base, the culture medium prepared is dispensed to 7 triangular flasks, 121 DEG C of sterilizing 30min, the respectively different numberings of the ring of picking 1 primary dcreening operation
In inoculation to triangular flask, mix after 25~30 DEG C accumulation culture 72h, determine each number spore count, germination percentage and
Enzymatic activity (basic protein enzyme activity, acid protease activity, neutral protease vigor and amylase activity), the result of measure
It is shown in Table 1.
The each numbering koji measurement result of table 1
As it can be seen from table 1 spore count highest, germination percentage and enzyme activity highest bacterial strain are the bacterial strains that numbering is Lp001,
By its freezing to China typical culture collection center preservation, address：Chinese Wuhan Wuhan Universitys, preservation date
On March 1st, 2017；Deposit number is CCTCC NO:M2017077；Classification And Nomenclature：Aspergillus oryzae HNCC 6.189, Latin literature
Name： Asporgillus orgzoeHNCC 6.189；Biological specimen viability is detected on March 11st, 2017 and completed, as a result
It is survival.
Embodiment 2：
Analysis for soybean powder, wheat-middlings, wheat bran, water are pressed 7.2:2:0.8:30 ratio is prepared, and adds the chlorine of gradient mass concentration
Change sodium solution, be prepared into the culture medium of the different mass fractions of salinity 12%, 14%, 16%, 18%, 20%, 22%, 24%,
Sterilize 30min in 121 DEG C, and the LP001 bacterial strains in embodiment 1 and aspergillus oryzae 3.951 are distinguished into the ring spore inoculating of picking 1 in training
Support in base, 72h is cultivated at 30 DEG C.The koji spore count of LP001 bacterial strains and aspergillus oryzae 3.951 in the present embodiment, germination percentage
And the contrast of enzymatic activity, the testing result of osmotic pressure is shown in Table 2.
Table 2：The contrasting detection result of LP001 bacterial strains and aspergillus oryzae 3.951 under different salinity
As can be seen that LP001 bacterial strains highest produced by the present invention is resistant to salinity up to 20%, when salt is dense from upper table 2
When degree increases to 22%, its koji spore count, germination percentage, enzyme activity are begun to decline, but are all significantly larger than starting strain aspergillus oryzae
3.951 measurement result.As a result show, the salt resistant character of LP001 bacterial strains of the present invention is much larger than starting strain aspergillus oryzae 3.951
Salt tolerance.
Embodiment 3：
By the LP001 bacterial strains in embodiment 1 on brewer's wort slant medium the generation of continuous passage culture 10, and by the 1st
Generation, the 3rd generation, the 5th generation, the 8th generation, the slant strains in the 10th generation make Triangle-botde koji, determine its spore count, germination percentage and enzyme
Activity, judges its genetic stability, and specific data are shown in Table 3.
Table 3：LP001 strain passage stability test results
From table 3 it can be seen that from the point of view of koji spore count, germination percentage and Enzyme assay result, the something lost of LP001 bacterial strains
Biography has good stability.
Embodiment 4：
A kind of high enzyme activity, the application process of aspergillus oryzae strain of high salt tolerant are：
7.2 portions of soybean, 2 parts of wheat-middlings, 0.8 part of wheat bran are weighed in parts by weight, by water after soybean is soaked 4 hours
Put only, take out soya bean, add the wheat bran weighed up, load boiling in normal pressure steamer, to steam to well-done without rotten, bean cotyledon is separated
To be suitable, then mixed with wheat-middlings, spread out and carry out being cooled to 25~30 DEG C, access high enzyme activity, the aspergillus oryzae of high salt tolerant
LP001 bacterial strains, inoculum concentration 0.2%, mix, in 30 DEG C of accumulation culture 72h, then song is made into.By the Cheng Quyu salt of making
Water is according to 1：3 mass ratio is added in sauce fermentation technique, and zymotechnique thereafter is consistent with former fermentation method, fermentation
150 days time, hair oil is filtered, soy sample, the detection of gas chromatography-mass spectrum (SDE/GC-MS) method are handled using distillation extraction
Every flavor of soy sauce changes of contents of its soy sauce made from starting strain aspergillus oryzae 3.951, while determine raw material and residue of soya
Total nitrogen content, calculate protein utilization, the results are shown in Table 4.
Table 4：The Indexs measure result of LP001 bacterial strains and soy sample made from aspergillus oryzae 3.951
From table 4, it can be seen that soy sauce and starting strain aspergillus oryzae is made in aspergillus oryzae LP001 bacterial strains produced by the present invention
Soy sauce made from 3.951 fermentations is compared, under conditions of fermentation period shortens one month, its amino-acid nitrogen, total acid, albumen
Matter utilization rate brings up to 89% from 80.02%, improves positive effect；The content of flavor substance is far above starting strain aspergillus oryzae
Soy sauce made from 3.951 fermentations, wherein alcohols material relative amount add 1.17% citronellol and 0.63% ethanol,
Aldehydes matter adds 0.19% 4-ethyl guaiacol, acid add 0.25% 2,4- adipic acids, 1.11%
Caproic acid and 1.57% syringic acid, Ester adds 3.75% ethyl acetate, 0.06% methyl stearate, aldehyde
Letones add 1.53% 5- methyl-2-furfurals, (the 2H)-furanone of 5.66%4- hydroxyl -2- ethyl -5- methyl -3,
1.09% 2,5- dimethyl-pyrazins and 0.23% 2- furfurals, above flavor substance mostly have typical paste flavor and peculiar
Flavor component, make flavor of soy sauce more abundant, pure and sweet.
Finally it should be noted that：The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；To the greatest extent
The present invention is described in detail with reference to the foregoing embodiments for pipe, it will be understood by those within the art that：It is still
Technical scheme described in foregoing embodiments can be modified, or which part technical characteristic is equally replaced
Change；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention technical scheme
Spirit and scope.
Claims (9)
1. aspergillus oryzae (Asporgillus orgzoe) bacterial strain of a kind of high enzyme activity, high salt tolerant, it is characterised in that deposit number is
CCTCC NO:M2017077。
2. the selection of the aspergillus oryzae strain of a kind of high enzyme activity, high salt tolerant, it is characterised in that comprise the following steps：
(1) starting strain is inoculated in 60~90h of culture on casein agar plating medium, selects the flat board to grow fine and enter
OK60Co- gamma-ray irradiation mutagenesis, Induced dosage choose 1~5kGy, put after having radiated and separated into desinfection chamber, continue to train
60~90h is supported, according to mycelial growth situation and bacterium colony transparent circle size, is filtered out using starting strain as control group eugonic
Bacterial strain.
(2) by the inoculation that step (1) screens on PDA slant mediums, after 25~30 DEG C of culture to sprouting periods,
With normal saline flushing slant medium, thalline is collected by centrifugation, adds physiological saline and obtains spore count as 106~107Individual/ml spore
Sub- suspension；
(3) take mutagens stoste to add and mutagens mother liquor is configured into buffer solution, in the spore suspension described in step (2)
With volume ratio 1：3~5 add mutagens mother liquor, are centrifuged after handling 60~120min at 25~30 DEG C, with buffer solution repeatedly
Rinse to reaction terminating；
(4) spore suspension after step (3) processing is respectively coated the casein agar flat board training in addition sodium chloride solution
Support and cultivated on base, according to mycelial growth situation and bacterium colony transparent circle size, it is vigorous to filter out growth using starting strain as control group
Bacterium colony, and be forwarded to immediately on wort agar inclined-plane culture medium, in 25~30 DEG C of cultures to ripe primary dcreening operation bacterial strain, then
It is standby to be placed in 4 DEG C of refrigerations；
(5) preferred culture medium is prepared by a certain percentage with analysis for soybean powder, wheat-middlings, wheat bran, water, the culture medium prepared is dispensed to three
Angle bottle, 121 DEG C of 25~30min of sterilizing, in primary dcreening operation inoculation to the triangular flask described in the ring step (4) of picking 1, is mixed after 25
~30 DEG C of accumulation 60~90h of culture, determine its spore count, germination percentage and enzymatic activity, spore count highest, germination percentage and enzyme activity highest
Bacterial strain be high enzyme activity, the aspergillus oryzae strain of high salt tolerant.
3. the selection of the aspergillus oryzae strain of a kind of high enzyme activity according to claim 2, high salt tolerant, it is characterised in that
Starting strain described in step (1) is aspergillus oryzae 3.951.
4. the selection of the aspergillus oryzae strain of a kind of high enzyme activity according to claim 2, high salt tolerant, it is characterised in that
Mutagens described in step (3) are ethylmethane sulfonates, and described buffer solution is the phosphate buffer that pH is 7.0~8.0.
5. the selection of the aspergillus oryzae strain of a kind of high enzyme activity according to claim 2, high salt tolerant, it is characterised in that
The initial mass fraction of sodium chloride solution described in step (4) is 10~14%, incremental gradient 2%, and final mass fraction is
22~26%.
6. the selection of the aspergillus oryzae strain of a kind of high enzyme activity according to claim 2, high salt tolerant, it is characterised in that
The recipe ingredient of casein agar plating medium described in step (1) (4) by weight for 15 parts of pancreatic digest of casein,
5 parts of soy meal papain digestion things, 5 parts of sodium chloride, 15 parts of agar, 1000 parts of distilled water；Described wort agar is oblique
The recipe ingredient of face culture medium is by weight 130 parts of fructus hordei germinatus leaching powders, 0.1 part of chloramphenicol, 5 parts of sodium chloride, 5.0,15 parts of agar
15.0th, 1000 parts of distilled water.
7. the selection of the aspergillus oryzae strain of a kind of high enzyme activity according to claim 2, high salt tolerant, it is characterised in that
The recipe ingredient of preferred culture medium described in step (5) is 7 parts of analysis for soybean powder, 2 parts of wheat-middlings, 1 part of wheat bran, 30 parts of distillations by weight
Water.
8. the selection of the aspergillus oryzae strain of a kind of high enzyme activity according to claim 2, high salt tolerant, it is characterised in that
Described high enzyme activity, the aspergillus oryzae strain of high salt tolerant are applied to high-salt diluted state fermentation soy brewage process.
A kind of 9. application process of the aspergillus oryzae strain of high enzyme activity, high salt tolerant, it is characterised in that including：
A, 7.2 portions of soybean, 2 parts of wheat-middlings, 0.8 part of wheat bran are weighed in parts by weight, by water after soybean is soaked 3~5 hours
Put only, take out soya bean, add the wheat bran weighed up, load boiling in normal pressure steamer, to steam to well-done without rotten, bean cotyledon is split up into
Suitably, then with wheat-middlings mixed, spread out and carry out being cooled to 25~30 DEG C, accessed high enzyme activity, the aspergillus oryzae strain of high salt tolerant, connect
Kind amount is 0.2%, is mixed, and cultivates 72h in 30 DEG C of accumulations, then song is made into.
B, by the Cheng Quyu salt solution of making according to 1：2~5 mass ratio is added in sauce fermentation technique, further preferably
Mass ratio be 1：3.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112574891A (en) * | 2020-12-01 | 2021-03-30 | 烟台欣和企业食品有限公司 | Aspergillus oryzae and application thereof |
| CN113373063A (en) * | 2021-06-29 | 2021-09-10 | 佛山市海天(高明)调味食品有限公司 | Aspergillus oryzae ZA175 and application thereof |
| CN117625422A (en) * | 2024-01-26 | 2024-03-01 | 龙牌食品股份有限公司 | Pichia burton strain L9-1 and application thereof |
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| CN104605308A (en) * | 2013-11-05 | 2015-05-13 | 孙德善 | Preparation method of soy sauce koji suitable for microalgal health liquid-state fermentation |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112574891A (en) * | 2020-12-01 | 2021-03-30 | 烟台欣和企业食品有限公司 | Aspergillus oryzae and application thereof |
| CN113373063A (en) * | 2021-06-29 | 2021-09-10 | 佛山市海天(高明)调味食品有限公司 | Aspergillus oryzae ZA175 and application thereof |
| CN117625422A (en) * | 2024-01-26 | 2024-03-01 | 龙牌食品股份有限公司 | Pichia burton strain L9-1 and application thereof |
| CN117625422B (en) * | 2024-01-26 | 2024-05-28 | 龙牌食品股份有限公司 | Pichia burton strain L9-1 and application thereof |
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Denomination of invention: Selection and application of a soy sauce strain with high enzyme activity and high salt tolerance Effective date of registration: 20221128 Granted publication date: 20210402 Pledgee: Hunan Tancheng Financing Guarantee Group Co.,Ltd. Pledgor: LONGPAI FOOD CO.,LTD. Registration number: Y2022980023965 |
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