CN109234186A - A kind of novel thermophilic lactic acid bacteria and its application in wet process corn soaking - Google Patents
A kind of novel thermophilic lactic acid bacteria and its application in wet process corn soaking Download PDFInfo
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- CN109234186A CN109234186A CN201810659554.0A CN201810659554A CN109234186A CN 109234186 A CN109234186 A CN 109234186A CN 201810659554 A CN201810659554 A CN 201810659554A CN 109234186 A CN109234186 A CN 109234186A
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- Prior art keywords
- lactic acid
- bacterial agent
- microbial bacterial
- freeze
- acid bacteria
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- 241000894006 Bacteria Species 0.000 title claims abstract description 88
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 66
- 240000008042 Zea mays Species 0.000 title claims abstract description 39
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 39
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 35
- 235000005822 corn Nutrition 0.000 title claims abstract description 35
- 239000004310 lactic acid Substances 0.000 title claims abstract description 33
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 33
- 238000002791 soaking Methods 0.000 title claims abstract description 18
- 238000000034 method Methods 0.000 title claims abstract description 16
- 244000005700 microbiome Species 0.000 claims abstract description 17
- 238000004321 preservation Methods 0.000 claims abstract description 7
- 241000193749 Bacillus coagulans Species 0.000 claims abstract description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 46
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 45
- 230000001580 bacterial effect Effects 0.000 claims description 43
- 230000000813 microbial effect Effects 0.000 claims description 43
- 239000000047 product Substances 0.000 claims description 42
- 239000000203 mixture Substances 0.000 claims description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 34
- 238000004108 freeze drying Methods 0.000 claims description 32
- 238000002360 preparation method Methods 0.000 claims description 32
- 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 30
- 238000000855 fermentation Methods 0.000 claims description 30
- 230000004151 fermentation Effects 0.000 claims description 30
- 239000008103 glucose Substances 0.000 claims description 30
- 239000012530 fluid Substances 0.000 claims description 21
- 235000015097 nutrients Nutrition 0.000 claims description 21
- 229920002261 Corn starch Polymers 0.000 claims description 19
- 239000008120 corn starch Substances 0.000 claims description 19
- 229940099112 cornstarch Drugs 0.000 claims description 19
- 229940041514 candida albicans extract Drugs 0.000 claims description 15
- 235000020183 skimmed milk Nutrition 0.000 claims description 15
- 239000012138 yeast extract Substances 0.000 claims description 15
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 14
- 239000001888 Peptone Substances 0.000 claims description 14
- 108010080698 Peptones Proteins 0.000 claims description 14
- 238000005119 centrifugation Methods 0.000 claims description 14
- 239000012153 distilled water Substances 0.000 claims description 14
- 235000019319 peptone Nutrition 0.000 claims description 14
- 230000001681 protective effect Effects 0.000 claims description 14
- FCBUKWWQSZQDDI-UHFFFAOYSA-N rhamnolipid Chemical compound CCCCCCCC(CC(O)=O)OC(=O)CC(CCCCCCC)OC1OC(C)C(O)C(O)C1OC1C(O)C(O)C(O)C(C)O1 FCBUKWWQSZQDDI-UHFFFAOYSA-N 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 12
- 239000000725 suspension Substances 0.000 claims description 11
- 239000000284 extract Substances 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 8
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 7
- 239000012267 brine Substances 0.000 claims description 7
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 7
- 239000002054 inoculum Substances 0.000 claims description 7
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 7
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 7
- 229940099596 manganese sulfate Drugs 0.000 claims description 7
- 239000011702 manganese sulphate Substances 0.000 claims description 7
- 235000007079 manganese sulphate Nutrition 0.000 claims description 7
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 7
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 7
- 229920000053 polysorbate 80 Polymers 0.000 claims description 7
- 239000001632 sodium acetate Substances 0.000 claims description 7
- 235000017281 sodium acetate Nutrition 0.000 claims description 7
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 235000015278 beef Nutrition 0.000 claims description 6
- 230000000630 rising effect Effects 0.000 claims description 5
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 5
- 239000001393 triammonium citrate Substances 0.000 claims description 5
- 235000011046 triammonium citrate Nutrition 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 3
- 230000008520 organization Effects 0.000 claims 1
- 239000002609 medium Substances 0.000 description 38
- 238000007654 immersion Methods 0.000 description 22
- 229920002472 Starch Polymers 0.000 description 17
- 239000008107 starch Substances 0.000 description 17
- 235000019698 starch Nutrition 0.000 description 17
- 239000000706 filtrate Substances 0.000 description 14
- 239000002068 microbial inoculum Substances 0.000 description 9
- 230000004083 survival effect Effects 0.000 description 8
- 238000007710 freezing Methods 0.000 description 7
- 230000008014 freezing Effects 0.000 description 7
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- 239000002002 slurry Substances 0.000 description 6
- RAHZWNYVWXNFOC-UHFFFAOYSA-N Sulphur dioxide Chemical compound O=S=O RAHZWNYVWXNFOC-UHFFFAOYSA-N 0.000 description 4
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 235000009973 maize Nutrition 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 235000005979 Citrus limon Nutrition 0.000 description 2
- 244000131522 Citrus pyriformis Species 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 235000015110 jellies Nutrition 0.000 description 2
- 239000008274 jelly Substances 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000004899 motility Effects 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- KVYRCBOUKXJXDK-UHFFFAOYSA-N 3,4-dimethylphenazine-1,2-diamine hydrochloride Chemical compound Cl.C1=CC=CC2=NC3=C(C)C(C)=C(N)C(N)=C3N=C21 KVYRCBOUKXJXDK-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 241000861996 Staphylococcus succinus Species 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 238000003916 acid precipitation Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 125000004122 cyclic group Chemical class 0.000 description 1
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
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- 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
- C12N1/205—Bacterial isolates
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B30/00—Preparation of starch, degraded or non-chemically modified starch, amylose, or amylopectin
- C08B30/04—Extraction or purification
- C08B30/042—Extraction or purification from cereals or grains
- C08B30/044—Extraction or purification from cereals or grains from corn or maize
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N1/04—Preserving or maintaining viable microorganisms
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- 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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Abstract
The present invention relates to a kind of novel thermophilic lactic acid bacteria and its application in wet process corn soaking, the culture presevation information of the thermophilic lactic acid bacteria: depositary institution's title: China Committee for Culture Collection of Microorganisms's common micro-organisms center;Depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica;Preservation date: on April 18th, 2018;Deposit number: CGMCC No.15623;Classification naming: bacillus coagulans Bacillus coagulans.
Description
Technical field
The invention belongs to microbial bacterial agent fields, and in particular to a kind of novel thermophilic lactic acid bacteria and its in wet process corn soaking
In application.
Background technique
Traditional handicraft cooperates with immersion with eo-acid (0.2% or so sulfurous acid solution) using old slurry in steeping tank, complete
Whole soaking process is included in new tinning plus old slurry, and the long corn of soaking time adds eo-acid, regular between tank and tank to follow
Ring.This cyclic process can make the solable matter in corn and soak keep a certain concentration poor, object soluble in corn
Matter, which soaks out, to be come.But the sulfur dioxide generated in traditional soaking process is the main component of acid rain, will cause environmental pollution, simultaneously
Sulfurous acid can also corrode equipment and pipeline, increase production cost.In the final product, the residual of sulfur dioxide will affect starch production
The quality of product and the utilization of by-product reduce economic benefit.Therefore, the soaking conditions of a new, alternative sulfurous acid are found
As the task of top priority.It needs multiple tank to connect in traditional tank switching soaking technology, is not suitable for the lower situation of cornstarch demand,
And tank switching process is cumbersome, increases production and equipment cost.In corn wet soaking process, thermophilic cream abundant in soak
Sour bacterium has played important function.Lactic acid bacteria has the ability of protein degradation matter, so that insoluble protein is changed into soluble protein, together
When the lactic acid that generates of lactobacillus-fermented and sulfurous acid synergistic effect can lead to cell wall in porous state, make to pass through originally
The high molecular weight protein enzyme and hydrone of corn epidermis can enter inside corn kernel, accelerate the water suction speed of corn kernel
Degree, promotes the separation of starch and protein in corn, to achieve the purpose that shorten soaking time.Add during corn soaking
Add thermophilic lactic acid bacteria bacterium solution, content of lactic acid bacteria in soak can be increased, gives full play to lactic acid bacteria effect, improve and impregnate effect.
But the limitations such as that there are storage lives is short for lactic acid bacterial liquid, stability is low and is not easy to transport.Therefore, one kind is developed to be avoided that using sulfurous
Acid can significantly improve corn wet simultaneously and impregnate the microbial bacterial agent of effect as the effective way for solving drawbacks described above.
Summary of the invention
The present invention provides a kind of thermophilic lactic acid bacteria LAB 0#, which is isolated from soak water of maize (old slurry).It is of the present invention
The culture presevation information of thermophilic lactic acid bacteria LAB 0#: depositary institution's title: China Committee for Culture Collection of Microorganisms is common
Microorganism center;Depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica;
Preservation date: on April 18th, 2018;Deposit number: CGMCC No.15623;Classification naming: bacillus coagulans Bacillus
coagulans。
Another embodiment of the present invention provides a kind of thermophilic lactic acid bacteria LAB 1#, and it is (old which is isolated from soak water of maize
Slurry).The culture presevation information of thermophilic lactic acid bacteria LAB 1# of the present invention: depositary institution's title: Chinese microorganism strain preservation
Administration committee's common micro-organisms center;Depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences
Institute of microbiology;Preservation date: on April 18th, 2018;Deposit number: CGMCC No.15624;Classification naming: amber grape
Coccus Staphylococcus succinus.
Another embodiment of the present invention provides a kind of microbial bacterial agent, it is characterised in that the function in the microbial bacterial agent
Microorganism is selected from one of thermophilic lactic acid bacteria LAB 0#, thermophilic lactic acid bacteria LAB 1# or two kinds.
Mentioned microorganism microbial inoculum further includes freeze drying protectant, and the freeze drying protectant is by following frozen-dried protective at being grouped as:
Rhamnolipid 100-300mg/L, glucose 80-120g/L, skimmed milk 120-150g/L, glycerol 12-15mL/L.
Another embodiment of the present invention provides a kind of microbial bacterial agent A, it is characterised in that microbial bacterial agent A is by thermophilic
Lactic acid bacteria LAB 0# and freeze drying protectant form, wherein viable bacteria amount >=3.6 × 10 of thermophilic lactic acid bacteria LAB 0#9/ g, the jelly
Dry protective agent is by following frozen-dried protective at being grouped as: rhamnolipid 200-300mg/L, glucose 80-120g/L, skimmed milk 120-
150g/L, glycerol 12-15mL/L.
The preparation method of another embodiment of the present invention offer mentioned microorganism microbial inoculum A, it is characterised in that including as follows
Step:
(1) preparation of seed liquor: thermophilic lactic acid bacteria LAB 0# is accessed in MRS fluid nutrient medium, at 50-55 DEG C, shaking table
Obtain seed liquor within culture 24 hours;
(2) preparation of bacterium mud: in the seed liquor access fermentation medium that step (1) is obtained, shaking table culture at 50-55 DEG C
After 24 hours, by fermentation material centrifugation, discard supernatant liquid, collect thallus, with brine to get arriving bacterium mud;
(3) preparation of microbial bacterial agent: mixing to obtain bacteria suspension for bacterium mud and freeze drying protectant that step (2) obtains, in-
It at 70--80 DEG C after pre-freeze 3-4h, is transferred to after being freeze-dried 20-24h in freeze drier, obtains bacterium powder, pack up to described micro-
Bacteria agent A.
The formula of MRS fluid nutrient medium described in step (1) is 10.0g containing peptone, ox in every liter of MRS fluid nutrient medium
Meat extract 5.0g, yeast extract 4.0g, glucose 20.0g, Tween-80 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, lemon
Three ammonium 2.0g of acid, magnesium sulfate 0.2g, manganese sulfate 0.05g, surplus are distilled water;Shaking speed is 100-120r/min;
The formula of fermentation medium is 10g containing glucose, yeast extract 10g, albumen in every liter of fermentation medium in step (2)
Peptone 10g, surplus are distilled water;Shaking speed is 100-120r/min;Seed liquor access fermentation medium in inoculum concentration be
5%;Centrifugal rotational speed is 5000r/min, centrifugation time 10-15min;
The volume ratio 1:1.5-2.0 of bacterium mud and freeze drying protectant, the freeze drying protectant are protected by following freeze-drying in step (3)
It protects into and is grouped as: rhamnolipid 200-300mg/L, glucose 80-120g/L, skimmed milk 120-150g/L, glycerol 12-15mL/
L。
Another embodiment of the present invention provides a kind of microbial bacterial agent B, it is characterised in that microbial bacterial agent B is by thermophilic
Lactic acid bacteria LAB 1# and freeze drying protectant form, wherein viable bacteria amount >=3.6 × 10 of thermophilic lactic acid bacteria LAB 1#9/ g, the jelly
Dry protective agent is by following frozen-dried protective at being grouped as: rhamnolipid 100-200mg/L, glucose 80-120g/L, skimmed milk 120-
150g/L, glycerol 12-15mL/L.
The preparation method of another embodiment of the present invention offer mentioned microorganism microbial inoculum B, it is characterised in that including as follows
Step:
(1) preparation of seed liquor: thermophilic lactic acid bacteria LAB 1# is accessed in MRS fluid nutrient medium, at 50-55 DEG C, shaking table
Obtain seed liquor within culture 24 hours;
(2) preparation of bacterium mud: in the seed liquor access fermentation medium that step (1) is obtained, shaking table culture at 50-55 DEG C
After 24 hours, by fermentation material centrifugation, discard supernatant liquid, collect thallus, with brine to get arriving bacterium mud;
(3) preparation of microbial bacterial agent: mixing to obtain bacteria suspension for bacterium mud and freeze drying protectant that step (2) obtains, in-
It at 70--80 DEG C after pre-freeze 3-4h, is transferred to after being freeze-dried 20-24h in freeze drier, obtains bacterium powder, pack up to described micro-
Bacteria agent B.
The formula of MRS fluid nutrient medium described in step (1) is 10.0g containing peptone, ox in every liter of MRS fluid nutrient medium
Meat extract 5.0g, yeast extract 4.0g, glucose 20.0g, Tween-80 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, lemon
Three ammonium 2.0g of acid, magnesium sulfate 0.2g, manganese sulfate 0.05g, surplus are distilled water;Shaking speed is 100-120r/min;
The formula of fermentation medium is 10g containing glucose, yeast extract 10g, albumen in every liter of fermentation medium in step (2)
Peptone 10g, surplus are distilled water;Shaking speed is 100-120r/min;Seed liquor access fermentation medium in inoculum concentration be
5%;Centrifugal rotational speed is 5000r/min, centrifugation time 10-15min;
The volume ratio 1:1.5-2.0 of bacterium mud and freeze drying protectant, the freeze drying protectant are protected by following freeze-drying in step (3)
It protects into and is grouped as: rhamnolipid 100-200mg/L, glucose 80-120g/L, skimmed milk 120-150g/L, glycerol 12-15mL/
L。
A kind of microbial bacterial agent composition, it is characterised in that the composition is by mentioned microorganism microbial inoculum A and mentioned microorganism
Microbial inoculum B composition;Wherein the mass ratio of the two is 1:1.0-1.5.
Another embodiment of the present invention provides mentioned microorganism microbial inoculum, mentioned microorganism microbial inoculum A and/or B, above-mentioned micro- life
Application of the object antimicrobial composition in wet process corn soaking.
Another embodiment of the present invention provides mentioned microorganism microbial inoculum, mentioned microorganism microbial inoculum A and/or B, above-mentioned micro- life
Application of the object antimicrobial composition in cornstarch extraction.
The present invention provides a kind of technique that wet process corn soaking extracts starch, it is characterised in that includes the following steps:
(1) corn, microbial bacterial agent, water are mixed, obtains immersion mixture after 12-20h is impregnated at 50-55 DEG C;
(2) by the immersion mixture that step (1) obtains be transferred in pulverizer carry out coarse crushing after, add microbial bacterial agent after
After continuous immersion 8-12h, progress is finely divided, and after successively crossing 50,250 meshes, filtrate stands overnight, is centrifugated, drying and to obtain corn
Starch.
Corn in step (1), microbial bacterial agent, water mass ratio be 1:0.01-0.03:3-5;The coarse crushing makes corn
Particle Breakage is at 5-8 valve;
The dosage of microbial bacterial agent is 0.5 times of the dosage of microbial bacterial agent in step (1) in step (2).
Compared with the prior art, the advantages of the present invention are as follows: (1) the present invention provides 2 kind novel thermophilic lactic acid bacterias, and
It is prepared into microbial bacterial agent, direct putting type bacteria agent is can be used as and is directly used in wet process corn soaking extraction starch, avoid people
Work adds lactic acid and sulfurous acid;(2) microbial bacterial agent A of the present invention and B, which is applied in combination, can be improved cornstarch yield, subtract simultaneously
Few soaking time, greatly improves the efficiency of wet process corn soaking;(3) when the present invention prepares microbial bacterial agent, one kind is selected
Have the rhamnolipid of very strong protective effect as one of protectant ingredient LAB 0# and 1# of the present invention, can significantly increase
Survival rate is lyophilized, survival rate is lyophilized 90% or more.
Specific embodiment
For the ease of a further understanding of the present invention, examples provided below has done more detailed description to it.But
It is that these embodiments are only not supposed to be a limitation to the present invention or implementation principle for better understanding invention, reality of the invention
The mode of applying is not limited to the following contents.
The screening of embodiment 1LAB 0#, LAB 1#
Soak water of maize (old slurry) 10mL is taken, is diluted to the 10 of original content respectively-1、10-2、10-3、10-4、10-5、10-6With
10-7Times, then 0.3mL is taken to be coated on MC plate respectively, (50 ± 2) DEG C culture is for 24 hours.Picking has the single colonie of molten calcium circle,
Flat lining out, separation, purify repeatedly.Isolate that molten calcium circle is big and unique bacterium colony access slant preservation, respectively number LAB
0#,LAB 1#.The soak water of maize (old slurry) is mentioned by Linghua Group Co., Ltd. (Jining, Shandong) production of corn starch workshop
For.
MC culture medium prescription: soy peptone 5.0g/L, beef extract powder 3.0g/L, yeast extract 3.0g/L, glucose
20.0g/L, lactose 20.0g/L, calcium carbonate 10.0g/L, agar 15.0g/L, dimethyl diaminophenazine chloride 0.05g/L, at 25 DEG C pH value be 6.0 ±
0.1。
The preparation of 2 microbial bacterial agent A of embodiment
(1) preparation of seed liquor: thermophilic lactic acid bacteria LAB 0# is accessed in MRS fluid nutrient medium, at 50-55 DEG C, 100r/
Obtain seed liquor within min shaking table culture 24 hours;Wherein the formula of MRS fluid nutrient medium is to contain albumen in every liter of MRS fluid nutrient medium
Peptone 10.0g, beef extract 5.0g, yeast extract 4.0g, glucose 20.0g, Tween-80 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate
5.0g, Triammonium citrate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, surplus are distilled water;
(2) preparation of bacterium mud: the seed liquor that step (1) is obtained is accessed in fermentation medium by 5% inoculum concentration, 50-
At 55 DEG C after 100r/min shaking table culture 24 hours, fermentation material is centrifuged (centrifugal rotational speed 5000r/min, centrifugation time
10min), discard supernatant liquid, collect thallus, with brine to get arriving bacterium mud;Wherein the formula of fermentation medium is every
Rising 10g containing glucose in fermentation medium, yeast extract 10g, peptone 10g, surplus is distilled water;
(3) preparation of microbial bacterial agent: 1:1.5 is mixed the bacterium mud and freeze drying protectant that step (2) is obtained by volume
Bacteria suspension is obtained, at -80 DEG C after pre-freeze 3h, is transferred to after being freeze-dried 20h in freeze drier, obtains bacterium powder, pack up to institute
State microbial bacterial agent A (hereinafter referred to as product a1);The freeze drying protectant is by following frozen-dried protective at being grouped as: rhamnolipid
200mg/L, glucose 120g/L, skimmed milk 120g/L, glycerol 15mL/L.The viable count 3.6 × 10 of product a19/ g, freezing are deposited
Motility rate 92.0%.
By bacterium mud that step (2) obtains, 1:1.5 mixes to obtain bacteria suspension by volume with freeze drying protectant, pre- at -80 DEG C
It after freezing 3h, is transferred to after being freeze-dried 20h in freeze drier, obtains bacterium powder, pack up to microbial bacterial agent (hereinafter referred to as product
a1-1);The freeze drying protectant is by following frozen-dried protective at being grouped as: glucose 120g/L, skimmed milk 120g/L, glycerol
15mL/L.The freeze survival rate 62.3% of product a1-1.
By bacterium mud that step (2) obtains, 1:1.5 mixes to obtain bacteria suspension by volume with freeze drying protectant, pre- at -80 DEG C
It after freezing 3h, is transferred to after being freeze-dried 20h in freeze drier, obtains bacterium powder, pack up to microbial bacterial agent (hereinafter referred to as product
a1-2);The freeze drying protectant is by following frozen-dried protective at being grouped as: rhamnolipid 50mg/L, glucose 120g/L, skimmed milk
120g/L, glycerol 15mL/L.The freeze survival rate 76.8% of product a1-2.
The preparation of 3 microbial bacterial agent A of embodiment
(1) preparation of seed liquor: thermophilic lactic acid bacteria LAB 0# is accessed in MRS fluid nutrient medium, at 50-55 DEG C, 120r/
Obtain seed liquor within min shaking table culture 24 hours;Wherein the formula of MRS fluid nutrient medium is to contain albumen in every liter of MRS fluid nutrient medium
Peptone 10.0g, beef extract 5.0g, yeast extract 4.0g, glucose 20.0g, Tween-80 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate
5.0g, Triammonium citrate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, surplus are distilled water;
(2) preparation of bacterium mud: the seed liquor that step (1) is obtained is accessed in fermentation medium by 5% inoculum concentration, 50-
At 55 DEG C after 120r/min shaking table culture 24 hours, fermentation material is centrifuged (centrifugal rotational speed 5000r/min, centrifugation time
15min), discard supernatant liquid, collect thallus, with brine to get arriving bacterium mud;Wherein the formula of fermentation medium is every
Rising 10g containing glucose in fermentation medium, yeast extract 10g, peptone 10g, surplus is distilled water;
(3) preparation of microbial bacterial agent: 1:2.0 is mixed the bacterium mud and freeze drying protectant that step (2) is obtained by volume
It obtains bacteria suspension to be transferred to after being freeze-dried for 24 hours in freeze drier at -70 DEG C after pre-freeze 4h, obtains bacterium powder, pack up to institute
State microbial bacterial agent A (hereinafter referred to as product a2);The freeze drying protectant is by following frozen-dried protective at being grouped as: rhamnolipid
300mg/L, glucose 80g/L, skimmed milk 150g/L, glycerol 12mL/L.The viable count 3.7 × 10 of product a29/ g, freezing survival
Rate 94.5%.
The preparation of 4 microbial bacterial agent B of embodiment
(1) preparation of seed liquor: thermophilic lactic acid bacteria LAB 1# is accessed in MRS fluid nutrient medium, at 50-55 DEG C, 100r/
Obtain seed liquor within min shaking table culture 24 hours;Wherein the formula of MRS fluid nutrient medium is to contain albumen in every liter of MRS fluid nutrient medium
Peptone 10.0g, beef extract 5.0g, yeast extract 4.0g, glucose 20.0g, Tween-80 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate
5.0g, Triammonium citrate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, surplus are distilled water;
(2) preparation of bacterium mud: the seed liquor that step (1) is obtained is accessed in fermentation medium by 5% inoculum concentration, 50-
At 55 DEG C after 100r/min shaking table culture 24 hours, fermentation material is centrifuged (centrifugal rotational speed 5000r/min, centrifugation time
10min), discard supernatant liquid, collect thallus, with brine to get arriving bacterium mud;Wherein the formula of fermentation medium is every
Rising 10g containing glucose in fermentation medium, yeast extract 10g, peptone 10g, surplus is distilled water;
(3) preparation of microbial bacterial agent: 1:1.5 is mixed the bacterium mud and freeze drying protectant that step (2) is obtained by volume
Bacteria suspension is obtained, at -80 DEG C after pre-freeze 3h, is transferred to after being freeze-dried 20h in freeze drier, obtains bacterium powder, pack up to institute
State microbial bacterial agent B (hereinafter referred to as product b1);The freeze drying protectant is by following frozen-dried protective at being grouped as: rhamnolipid
100mg/L, glucose 120g/L, skimmed milk 120g/L, glycerol 15mL/L.The viable count 4.0 × 10 of product b19/ g, freezing are deposited
Motility rate 91.2%.
By bacterium mud that step (2) obtains, 1:1.5 mixes to obtain bacteria suspension by volume with freeze drying protectant, pre- at -80 DEG C
It after freezing 3h, is transferred to after being freeze-dried 20h in freeze drier, obtains bacterium powder, pack up to microbial bacterial agent (hereinafter referred to as product
b1-1);The freeze drying protectant is by following frozen-dried protective at being grouped as: glucose 120g/L, skimmed milk 120g/L, glycerol
15mL/L.The freeze survival rate 63.1% of product b1-1.
By bacterium mud that step (2) obtains, 1:1.5 mixes to obtain bacteria suspension by volume with cryoprotector, pre- at -80 DEG C
It after freezing 3h, is transferred to after being freeze-dried 20h in freeze drier, obtains bacterium powder, pack up to microbial bacterial agent (hereinafter referred to as product
b1-2);The freeze drying protectant is by following frozen-dried protective at being grouped as: rhamnolipid 50mg/L, glucose 120g/L, skimmed milk
120g/L, glycerol 15mL/L.The freeze survival rate 79.3% of product b1-2.
The preparation of 5 microbial bacterial agent B of embodiment
(1) preparation of seed liquor: thermophilic lactic acid bacteria LAB 1# is accessed in MRS fluid nutrient medium, at 50-55 DEG C, 120r/
Obtain seed liquor within min shaking table culture 24 hours;Wherein the formula of MRS fluid nutrient medium is to contain albumen in every liter of MRS fluid nutrient medium
Peptone 10.0g, beef extract 5.0g, yeast extract 4.0g, glucose 20.0g, Tween-80 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate
5.0g, Triammonium citrate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, surplus are distilled water;
(2) preparation of bacterium mud: the seed liquor that step (1) is obtained is accessed in fermentation medium by 5% inoculum concentration, 50-
At 55 DEG C after 120r/min shaking table culture 24 hours, fermentation material is centrifuged (centrifugal rotational speed 5000r/min, centrifugation time
15min), discard supernatant liquid, collect thallus, with brine to get arriving bacterium mud;Wherein the formula of fermentation medium is every
Rising 10g containing glucose in fermentation medium, yeast extract 10g, peptone 10g, surplus is distilled water;
(3) preparation of microbial bacterial agent: 1:2.0 is mixed the bacterium mud and freeze drying protectant that step (2) is obtained by volume
It obtains bacteria suspension to be transferred to after being freeze-dried for 24 hours in freeze drier at -70 DEG C after pre-freeze 4h, obtains bacterium powder, pack up to institute
State microbial bacterial agent B (hereinafter referred to as product b2);The freeze drying protectant is by following frozen-dried protective at being grouped as: rhamnolipid
200mg/L, glucose 80g/L, skimmed milk 150g/L, glycerol 12mL/L.The freeze survival rate 93.6% of product b2.
Embodiment 6
(1) corn (10kg), product a1 (300g), water (30kg) are mixed, must be impregnated after impregnating 20h at 50-55 DEG C
Mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product a1 (150g) continues after impregnating 12h, and progress is finely divided, and after successively crossing 50,250 meshes, filtrate is stood overnight, centrifugation divides
From, dry cornstarch, starch yield 70.2%.
Embodiment 7
(1) corn (10kg), product a1 (300g), water (30kg) are mixed, must be impregnated after impregnating 12h at 50-55 DEG C
Mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product a1 (150g) continues after impregnating 8h, carries out finely divided, after successively crossing 50,250 meshes, filtrate stands overnight, is centrifugated,
Dry cornstarch, starch yield 62.3%.
Embodiment 8
(1) corn (10kg), product a1 (100g), water (30kg) are mixed, must be impregnated after impregnating 12h at 50-55 DEG C
Mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product a1 (50g) continues after impregnating 8h, carries out finely divided, after successively crossing 50,250 meshes, filtrate stands overnight, is centrifugated,
Dry cornstarch, starch yield 54.7%.
Embodiment 9
(1) corn (10kg), product a2 (300g), water (50kg) are mixed, must be impregnated after impregnating 20h at 50-55 DEG C
Mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product a2 (150g) continues after impregnating 12h, and progress is finely divided, and after successively crossing 50,250 meshes, filtrate is stood overnight, centrifugation divides
From, dry cornstarch, starch yield 70.5%.
Embodiment 10
(1) corn (10kg), product b1 (300g), water (30kg) are mixed, must be impregnated after impregnating 20h at 50-55 DEG C
Mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product b1 (150g) continues after impregnating 12h, and progress is finely divided, and after successively crossing 50,250 meshes, filtrate is stood overnight, centrifugation divides
From, dry cornstarch, starch yield 70.7%.
Embodiment 11
(1) corn (10kg), product b1 (300g), water (30kg) are mixed, must be impregnated after impregnating 12h at 50-55 DEG C
Mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product b1 (150g) continues after impregnating 8h, carries out finely divided, after successively crossing 50,250 meshes, filtrate stands overnight, is centrifugated,
Dry cornstarch, starch yield 63.5%.
Embodiment 12
(1) corn (10kg), product b1 (100g), water (30kg) are mixed, must be impregnated after impregnating 12h at 50-55 DEG C
Mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product b1 (50g) continues after impregnating 8h, carries out finely divided, after successively crossing 50,250 meshes, filtrate stands overnight, is centrifugated,
Dry cornstarch, starch yield 55.2%.
Embodiment 13
(1) corn (10kg), product b2 (300g), water (50kg) are mixed, must be impregnated after impregnating 20h at 50-55 DEG C
Mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product b2 (150g) continues after impregnating 12h, and progress is finely divided, and after successively crossing 50,250 meshes, filtrate is stood overnight, centrifugation divides
From, dry cornstarch, starch yield 70.6%.
Embodiment 14
(1) corn (10kg), product a1 (50g), product b1 (50g), water (30kg) are mixed, is impregnated at 50-55 DEG C
Immersion mixture is obtained after 12h;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product a1 (25g), product b1 (25g) continue after impregnating 8h, and progress is finely divided, and after successively crossing 50,250 meshes, filtrate was stood
Night, centrifuge separation, dry cornstarch, starch yield 72.1%.
Embodiment 15
(1) corn (10kg), product a1 (40g), product b2 (60g), water (30kg) are mixed, is impregnated at 50-55 DEG C
Immersion mixture is obtained after 12h;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and add after coarse crushing is broken into 5-8 valve
Product a1 (20g), product b2 (30g) continue after impregnating 8h, and progress is finely divided, and after successively crossing 50,250 meshes, filtrate was stood
Night, centrifuge separation, dry cornstarch, starch yield 71.9%.
Embodiment 16
(1) corn (10kg) is mixed with water (30kg), accesses the LAB 0# of 5% amount, after impregnating 20 at 50-55 DEG C
Immersion mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and is carried out after coarse crushing is broken into 5-8 valve, then connect
The LAB 0# (i.e. 2.5%) for entering half amount continues after impregnating 12h, and progress is finely divided, and after successively crossing 50,250 meshes, filtrate is stood
Overnight, centrifuge separation, dry cornstarch, starch yield 23.5%.
Embodiment 17
(1) corn (10kg) is mixed with water (30kg), accesses the LAB 1# of 5% amount, after impregnating 20 at 50-55 DEG C
Immersion mixture;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and is carried out after coarse crushing is broken into 5-8 valve, then connect
The LAB 1# (i.e. 2.5%) for entering half amount continues after impregnating 12h, and progress is finely divided, and after successively crossing 50,250 meshes, filtrate is stood
Overnight, centrifuge separation, dry cornstarch, starch yield 24.7%.
Embodiment 18
(1) corn (10kg) is mixed with water (30kg), is sequentially ingressed into the LAB 0# of 2.5% amount and the LAB of 2.5% amount
1# obtains immersion mixture after impregnating 20 at 50-55 DEG C;
(2) immersion mixture that step (1) obtains is transferred in pulverizer and is carried out after coarse crushing is broken into 5-8 valve, then connect
Enter half amount LAB 0# and LAB 1# (i.e. each 1.25%) continue impregnate 12h after, carry out it is finely divided, successively cross 50,250 meshes
Afterwards, filtrate stands overnight, is centrifugated, drying and to obtain cornstarch, starch yield 30.2%.
Claims (10)
1. a kind of thermophilic lactic acid bacteria LAB 0#, it is characterised in that the culture presevation information of the thermophilic lactic acid bacteria LAB 0#: preservation
Organization: China Committee for Culture Collection of Microorganisms's common micro-organisms center;Depositary institution address: Chaoyang District, Beijing City
The institute 3 of North Star West Road 1, Institute of Microorganism, Academia Sinica;Preservation date: on April 18th, 2018;Deposit number: CGMCC
No.15623;Classification naming: bacillus coagulans Bacillus coagulans.
2. a kind of microbial bacterial agent A, it is characterised in that microbial bacterial agent A is by thermophilic lactic acid bacteria LAB described in claim 1
0# and freeze drying protectant composition.
3. microbial bacterial agent A as claimed in claim 2, it is characterised in that the freeze drying protectant is by following frozen-dried protective at grouping
At: rhamnolipid 200-300mg/L, glucose 80-120g/L, skimmed milk 120-150g/L, glycerol 12-15mL/L.
4. the preparation method of the described in any item microbial bacterial agent A of claim 2-3, it is characterised in that include the following steps:
(1) preparation of seed liquor: thermophilic lactic acid bacteria LAB 0# is accessed in MRS fluid nutrient medium, at 50-55 DEG C, shaking table culture
Obtain seed liquor within 24 hours;
(2) preparation of bacterium mud: in the seed liquor access fermentation medium that step (1) is obtained, shaking table culture 24 is small at 50-55 DEG C
Shi Hou by fermentation material centrifugation, discards supernatant liquid, collect thallus, with brine to get arriving bacterium mud;
(3) preparation of microbial bacterial agent: bacterium mud and freeze drying protectant that step (2) obtains are mixed into obtain bacteria suspension, in -70--80
It at DEG C after pre-freeze 3-4h, is transferred to after being freeze-dried 20-24h in freeze drier, obtains bacterium powder, pack up to the microbial bacteria
Agent A.
5. preparation method as claimed in claim 4, it is characterised in that the formula of MRS fluid nutrient medium described in step (1) is every
Rise 10.0g containing peptone, beef extract 5.0g, yeast extract 4.0g, glucose 20.0g, Tween-80 in MRS fluid nutrient medium
1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, Triammonium citrate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, surplus are
Distilled water.
6. the described in any item preparation methods of claim 4-5, it is characterised in that the formula of fermentation medium is every in step (2)
Rising 10g containing glucose in fermentation medium, yeast extract 10g, peptone 10g, surplus is distilled water;Seed liquor accesses fermented and cultured
Inoculum concentration in base is 5%;Centrifugal rotational speed is 5000r/min, centrifugation time 10-15min.
7. the described in any item preparation methods of claim 4-6, it is characterised in that the body of bacterium mud and freeze drying protectant in step (3)
Product is than 1:1.5-2.0, and the freeze drying protectant is by following frozen-dried protective at being grouped as: rhamnolipid 200-300mg/L, glucose
80-120g/L, skimmed milk 120-150g/L, glycerol 12-15mL/L.
8. the described in any item preparation methods of claim 4-7, it is characterised in that shaking speed is 100- in step (1), (2)
120r/min。
9. application of the described in any item microbial bacterial agent A of claim 2-3 in wet process corn soaking.
10. application of the described in any item microbial bacterial agent A of claim 2-3 in cornstarch extraction.
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