CN106947703A - One plant of rhizopus stolonifer and its application - Google Patents
One plant of rhizopus stolonifer and its application Download PDFInfo
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- CN106947703A CN106947703A CN201710356131.7A CN201710356131A CN106947703A CN 106947703 A CN106947703 A CN 106947703A CN 201710356131 A CN201710356131 A CN 201710356131A CN 106947703 A CN106947703 A CN 106947703A
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- glutinous rice
- bacterial strain
- rhizopus
- yeast
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- 241000235546 Rhizopus stolonifer Species 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 22
- 230000001580 bacterial effect Effects 0.000 claims abstract description 21
- 235000019991 rice wine Nutrition 0.000 claims abstract description 21
- 241000209094 Oryza Species 0.000 claims abstract description 18
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 18
- 235000009566 rice Nutrition 0.000 claims abstract description 18
- 241000894006 Bacteria Species 0.000 claims abstract description 14
- 238000000855 fermentation Methods 0.000 claims abstract description 14
- 230000004151 fermentation Effects 0.000 claims abstract description 14
- 241000196324 Embryophyta Species 0.000 claims abstract description 13
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 11
- 230000008569 process Effects 0.000 claims abstract description 10
- 238000012216 screening Methods 0.000 claims abstract description 10
- 239000001963 growth medium Substances 0.000 claims abstract description 9
- 238000004321 preservation Methods 0.000 claims abstract description 6
- 241000235527 Rhizopus Species 0.000 claims abstract description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 22
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 239000002609 medium Substances 0.000 claims description 14
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 239000003643 water by type Substances 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 6
- 238000002955 isolation Methods 0.000 claims description 6
- 238000011218 seed culture Methods 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 238000007710 freezing Methods 0.000 claims description 5
- 230000008014 freezing Effects 0.000 claims description 5
- 235000011187 glycerol Nutrition 0.000 claims description 5
- 238000005360 mashing Methods 0.000 claims description 5
- 239000000243 solution Substances 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- 229920002472 Starch Polymers 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 238000009835 boiling Methods 0.000 claims description 4
- 238000004140 cleaning Methods 0.000 claims description 4
- 235000009508 confectionery Nutrition 0.000 claims description 4
- 238000001816 cooling Methods 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims description 4
- 239000012092 media component Substances 0.000 claims description 4
- 235000015097 nutrients Nutrition 0.000 claims description 4
- 235000019698 starch Nutrition 0.000 claims description 4
- 239000008107 starch Substances 0.000 claims description 4
- 238000012360 testing method Methods 0.000 claims description 4
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- 238000010790 dilution Methods 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 claims description 3
- 230000002538 fungal effect Effects 0.000 claims description 3
- 244000005700 microbiome Species 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 229930182555 Penicillin Natural products 0.000 claims description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 2
- 239000004570 mortar (masonry) Substances 0.000 claims description 2
- 229940049954 penicillin Drugs 0.000 claims description 2
- 239000000725 suspension Substances 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims 2
- 229910052801 chlorine Inorganic materials 0.000 claims 2
- 239000000460 chlorine Substances 0.000 claims 2
- 238000007598 dipping method Methods 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 10
- 108020004463 18S ribosomal RNA Proteins 0.000 abstract description 6
- 238000006243 chemical reaction Methods 0.000 abstract description 4
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- 239000000758 substrate Substances 0.000 abstract description 3
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- 235000014101 wine Nutrition 0.000 description 10
- 240000004638 Dendrobium nobile Species 0.000 description 5
- 241000593344 Rhizopus microsporus Species 0.000 description 4
- 240000005384 Rhizopus oryzae Species 0.000 description 4
- 235000013752 Rhizopus oryzae Nutrition 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 230000003637 steroidlike Effects 0.000 description 4
- 241001523681 Dendrobium Species 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- HDXIQHTUNGFJIC-UHFFFAOYSA-N (25R)-spirost-5-en-3beta-ol 3-O-<O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranoside> Natural products O1C2(OCC(C)CC2)C(C)C(C2(CCC3C4(C)CC5)C)C1CC2C3CC=C4CC5OC1OC(CO)C(O)C(O)C1OC1OC(C)C(O)C(O)C1O HDXIQHTUNGFJIC-UHFFFAOYSA-N 0.000 description 2
- VNONINPVFQTJOC-RXEYMUOJSA-N Collettiside III Natural products O([C@@H]1[C@@H](O)[C@H](O[C@H]2[C@H](O)[C@H](O)[C@@H](O)[C@H](C)O2)[C@H](CO)O[C@@H]1O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]5[C@H](C)[C@@]6(O[C@H]5C4)OC[C@H](C)CC6)CC3)CC=2)CC1)[C@H]1[C@H](O)[C@H](O)[C@@H](O)[C@H](C)O1 VNONINPVFQTJOC-RXEYMUOJSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 102000006995 beta-Glucosidase Human genes 0.000 description 2
- 108010047754 beta-Glucosidase Proteins 0.000 description 2
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 2
- 229960005091 chloramphenicol Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
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- VNONINPVFQTJOC-ZGXDEBHDSA-N dioscin Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@H](O)[C@@H](O)[C@H](C)O1)O)O[C@@H]1CC2=CC[C@H]3[C@@H]4C[C@H]5[C@@H]([C@]4(CC[C@@H]3[C@@]2(C)CC1)C)[C@@H]([C@]1(OC[C@H](C)CC1)O5)C)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O VNONINPVFQTJOC-ZGXDEBHDSA-N 0.000 description 2
- CJNUQCDDINHHHD-APRUHSSNSA-N dioscin Natural products C[C@@H]1CC[C@@]2(OC1)O[C@H]3C[C@H]4[C@@H]5CC=C6C[C@H](CC[C@@H]6[C@H]5CC[C@]4(C)[C@H]3[C@@H]2C)O[C@@H]7O[C@H](CO)[C@@H](O[C@@H]8O[C@@H](C)[C@H](O)[C@@H](O)[C@H]8O)[C@H](O)[C@H]7O[C@@H]9O[C@@H](C)[C@H](O)[C@@H](O)[C@H]9O CJNUQCDDINHHHD-APRUHSSNSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
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- 239000000203 mixture Substances 0.000 description 2
- VNONINPVFQTJOC-UHFFFAOYSA-N polyphyllin III Natural products O1C2(OCC(C)CC2)C(C)C(C2(CCC3C4(C)CC5)C)C1CC2C3CC=C4CC5OC(C(C1O)OC2C(C(O)C(O)C(C)O2)O)OC(CO)C1OC1OC(C)C(O)C(O)C1O VNONINPVFQTJOC-UHFFFAOYSA-N 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- LHNVKVKZPHUYQO-SRWWVFQWSA-N 16-alpha,17-Epoxypregn-4-ene-3,20-dione Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1C[C@H]3O[C@@]3(C(=O)C)[C@@]1(C)CC2 LHNVKVKZPHUYQO-SRWWVFQWSA-N 0.000 description 1
- TVEXGJYMHHTVKP-UHFFFAOYSA-N 6-oxabicyclo[3.2.1]oct-3-en-7-one Chemical compound C1C2C(=O)OC1C=CC2 TVEXGJYMHHTVKP-UHFFFAOYSA-N 0.000 description 1
- 244000027711 Brettanomyces bruxellensis Species 0.000 description 1
- 235000000287 Brettanomyces bruxellensis Nutrition 0.000 description 1
- 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 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000011837 pasties Nutrition 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000015096 spirit Nutrition 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Classifications
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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
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/845—Rhizopus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention belongs to technical field of bioengineering, one plant of rhizopus stolonifer and its application are specifically disclosed, by screening and 18S rRNA sequence analyses, isolated rhizopus strains rhizopus stolonifer bacterium is accredited as, its preservation is marked as CGMCC NO.13767.Bacterial strain of the present invention is screened using sticky rice juice culture medium to rhizopus, the bacterial strain screened more adapts to the working condition for the fermentation that glutinous rice is substrate, by adding the bacterial strain with efficiently saccharifying ability, be conducive to improving saccharification conversion ratio, be conducive to improving the raw material availability and production efficiency of glutinous rice wine brewage process.
Description
Technical field
The invention belongs to technical field of biological fermentation, more specifically, more particularly to one plant of rhizopus stolonifer screening technique
With the application of rhizopus stolonifer.
Background technology
China is the native place of wine, is also the cradle of spirits culture, is one of earliest country of making wine in the world.The wine of wine
Make, in the existing suitable long history of China.In Chinese thousands of years development of civilization history, the development of wine and culture is substantially
Synchronous progress.Glutinous rice wine is favored by consumers in general, is fermented for glutinous rice wine distiller's yeast, head mold is the main saccharification of distiller's yeast
Bacterium, yeast is main zymophyte, therefore distiller's yeast quality good or not, depending on wherein head mold and yeast species quality and quantity it is many
It is few.Head mold kind in different distiller's yeasts is different, and saccharifying power is variant greatly.Head mold of the same race is made distiller's yeast because of inoculum concentration, incubation time
With the difference of culture environment etc., saccharifying power is also big variant.The fermenting power of distiller's yeast, the activity of yeast also have similar with saccharifying power
Problem.
Patent of invention " Rhizopus microsporus C97102734 and application " (application number：CN201610054903.7 one plant) is disclosed
Rhizopus microsporus (Rhizopus microsporus) C97102734, carries out liquefaction to the stem of noble dendrobium using Rhizopus microsporus and prepares stem of noble dendrobium liquid
Change liquid, using enzymolysis, fermentation or the two combination by the way of, the dendrobium polysaccharide insoluble in wine is transformed into be dissolved in wine the stem of noble dendrobium it is low
Glycan, solves dendrobium polysaccharide insoluble in wine, it is impossible to the technical barrier of qualified wine is produced with the stem of noble dendrobium, obtained dendrobium wine has good
Good healthcare function, realizes stem of noble dendrobium deep processing, its added value is greatly improved.One plant of production beta-glucosidase of patent of invention "
Enzyme head mold and its enzymatic production method " (application number：CN201510731434.3 a kind of rhizopus stolonifer) is disclosed
(Rhizopusstolonifer) TP-02, deposit number is：CGMCCNo.11119, the rhizopus stolonifer
(Rhizopusstolonifer) TP-02 is screened from the rotten wood of Mount Huang and obtained, and the patent, which is also disclosed, utilizes rhizopus stolonifer
(Rhizopusstolonifer) method that TP-02 produces rhizopus stolonifer beta-glucosidase, and to the enzymatic property of institute's producing enzyme
Analyzed.Patent of invention " head mold of food fermentation and application " (application number：CN201410676014.5 one) is disclosed
The head mold of food fermentation and application are planted, the patent of invention bacterial strain is Rhizopus oryzae, isolated three plants from sweet rice wine, respectively
For Rhizopus oryzae zsm-003, zsm-004 and head mold zsm-005, fermented suitable for rice food and sweet rice wine；The patent is also disclosed
A kind of complex microbial inoculum of food fermentation, includes Rhizopus oryzae zsm-003, Rhizopus oryzae zsm-004 and Karst aroma
Yeast (Brettanomyces custersii) ZSM-001, the composite ferment strain survival rate reaches more than 35%, production
Product sweet rice wine in carbohydrate composition relatively enrich, product flavour is strong, in good taste, tool is higher delicious amino acid and must amino
Acid.A kind of patent of invention " bread mold and its application technology " (application number：CN201410136456.0 a kind of high yield steroid) is disclosed
Body fermented black rhizopus strains and its application technology of steroidal compounds fermentation, the bacterial strain preserving number is：CGMCC No.7687, it is main
It is used for the conversion process that steroidal compounds (Dioscin) produce plant hormone for raw material, with steroidal intermediate 16ALPHA,17ALPHA-epoxyprogesterone
Steroidal intermediate mold oxide is produced for substrate, there is production mold oxide high conversion rate to be reached up to 71%, high income
100%, and biological character is more stable, is that the plant steroid hormone production using Dioscin as raw material improves yield, drop
Low production cost provides technical support.
The content of the invention
The invention aims to improve the fermentation efficiency of tradition glutinous rice wine production method, one plant of saccharification capability is filtered out
Strong rhizopus stolonifer.
To achieve the above object, the present invention provides following technical scheme：One plant of rhizopus stolonifer and its application, including following step
Suddenly：
One plant of rhizopus stolonifer bacterium, the rhizopus stolonifer is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms
Center, its deposit number is CGMCC NO.13767；Preservation date is：On March 27th, 2017；Depositary institution is entitled：China
Microbiological Culture Collection administration committee common micro-organisms center；Preservation address is：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number, Institute of Microorganism, Academia Sinica；Classification And Nomenclature is：Rhizopus stolonifer Cy3, Rhizopus stoloniferCy3.This hair
It is bright that identification is classified to bacterial strain by 18S rRNA sequence methods.The DNA of this bacterium is extracted, 18S rRNA sequence is expanded,
Contrasted with reference rhizopus stolonifer bacterial strain, homology is 98%~100%.
Rhizopus stolonifer bacterial strain of the present invention is screened from glutinous rice wine distiller's yeast and obtained, the Mashing process for glutinous rice wine.
Distiller's yeast of the present invention is tradition glutinous rice wine solid distiller's yeast, is provided by Dragon Palace liquid Wine Co., Ltd of Hunan Province.
The screening technique of bacterial strain of the present invention is：Glutinous rice wine solid distiller's yeast 1.0g accurately is weighed, is put into sterile mortar, is added
10mL sterilized waters are ground, and wear into pasty state, are then added 40mL sterilized waters and are stirred, obtain distiller's yeast suspension, use oese
The ring of picking one is applied in sterile isolation medium in isolation medium with spread plate, 30 DEG C of culture 15h；In aseptic condition
Under, with the fungal strain that oese picking is maximum, access is equipped with the 25mL sterile test tubes of 20mL sterilized waters, is stirred dilute
Release, with the ring of oese picking one in single bacterium colony screening and culturing medium, be applied to spread plate in sterile single bacterium colony screening and culturing medium,
30 DEG C of culture 36h；Aseptically, with the maximum fungal strain of oese picking, 25mL of the access equipped with 20mL sterilized waters
In sterile test tube, dilution is stirred, 1mL is drawn in 1L seed culture mediums, 30 DEG C, under the conditions of 150rpm, 25h is cultivated, makees
It is standby for seed liquor, use -80 DEG C of ultralow temperature glycerine freezing pipe preserving process preservation seed liquors.
The present invention is so nutrient media components and its processing method：
Sterile isolation medium：Sticky rice juice 300mL, chloramphenicol 0.1g/L, agar 15.0g/L；Sterilizing methods are：It is accurate to claim
Culture medium each component is taken, heating for dissolving adjusts pH to 6.0 in 700ml distilled water with 0.1mol/L Hcl and NaOH solution, point
Triangular flask is filled, 121 DEG C of autoclavings 30 minutes take out, aseptically cooled down, be down flat plate.
Single bacterium colony screening and culturing medium：Sticky rice juice 300mL, chloramphenicol 0.1g/L, agar 15.0g/L, penicillin 80U/mL；Go out
Bacterium method is：Accurate weighing culture medium each component, heating for dissolving is molten with 0.1mol/L Hcl and NaOH in 700ml distilled water
Liquid adjusts pH to 6.0, dispenses triangular flask, and 121 DEG C of autoclavings 30 minutes take out, aseptically cooled down, be down flat plate..
Seed culture medium：Sticky rice juice 300mL, adds 700ml distilled water, and pH is adjusted with 0.1mol/L Hcl and NaOH solution
To 6.0.
Fermentation medium：Content of starch is more than 70% glutinous rice, by cleaning, steeped overnight, boiling 25min, after cooling,
The water of equal quality is added, it is stand-by.
The Mashing process that the present invention is used for glutinous rice wine production to institute's bacterium is illustrated：Take 1 pipe ultralow temperature glycerine
The strain that freezing pipe is preserved, puts room-temperature dissolution, pours into 1Kg fermentation mediums, by stirring, beats nest, is issued in 30 DEG C of conditions
Ferment 65h, by 4 layers of filtered through gauze, surveys the total pol of mash.
The technique effect and advantage of the present invention：One plant of rhizopus stolonifer and its application that the present invention is provided, with tradition glutinous rice wine
Brewing method is compared, and bacterial strain of the present invention is screened using sticky rice juice culture medium to head mold, and the bacterial strain screened more adapts to glutinous
Rice is the working condition of the fermentation of substrate, by adding the bacterial strain with efficiently saccharifying ability, is conducive to improving saccharification conversion ratio,
Be conducive to improving the raw material availability and production efficiency of glutinous rice wine brewage process.
Embodiment
Technical scheme is elaborated further below with reference to embodiment.
Embodiment 1：The separation and identification of rhizopus stolonifer bacterial strain
(1) rhizopus stolonifer bacterial strain of the present invention is isolated, the solid distiller's yeast from tradition glutinous rice wine solid distiller's yeast
There is provided by Dragon Palace liquid Wine Co., Ltd of Hunan Province.The seed culture condition of the bacterial strain is：Take the strain of a superfreeze
Glycerol tube, thaws at room temperature, is transferred to 300mL seed culture mediums, under the conditions of 30 DEG C, 150rpm, shaking table culture 25h；The training
Supporting base composition is：Sticky rice juice 300mL, adds 700ml distilled water, pH value 6.0.
(2) taxonomic identification is carried out to bacterial strain of the present invention using 18S rRNA gene sequencing methods.
18S rRNA sequencings commission Shanghai Sheng Gong bioengineering limited company is measured, after measured, the bacterium
The 18S rRNA sequences of strain are as follows：
AGTCATATGCTTGTCTCAAAGATTAAGCCATGCATGTCTAAGTATAAATAACTTTTATATTGTGAAACT
GCGAATGGCTCATTAAATCAGTTATGATCTACGTGACAAATTCTTTACTACTTGGATAACCGTGGTAATTCTAGAGC
TAATACATGCAAAAAAGCCCTGACTTACGAAGGGGTGCACTTATTAGATAAAATCAACGCGGGGTAAAACCTGTTTC
TTGGTGAATCATAATAATTAAGCGGATCGCATAGCCTTGTGCTGGCGACGGTCCACTCGATTTTCTGCCCTATCATG
GTTGAGATTGTAAGATAGAGGCTTACAATGCCTACAACGGGTAACGGGGAATTAGGGTTCGATTCCGGAGAGGGAGC
CTGAGAAACGGCTACCACATCCAAGGAAGGCAGCAGGCGCGCAAATTACCCAATCCCGACACGGGGAGGTAGTGACA
ATACATAACAATGCAGGGCCTTTAAGGTCTTGCAATTGGAATGAGTACAATTTAAATCCCTTAACGAGGATCAATTG
GAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATTCCAGCTCCAATAGCGTATATTAAAGTTGTTGCAGTTAAAACGT
CCGTAGTCAAACTTTAGTCTTACTGGCGTAGTGGCCTGGTCTTCATTGACCAAGCTCATTGCTGCCGGAGACTCCAT
GTTCATTAGGGGAAAATCTCTAGGGGTTTTTTTCTTATTTGAACAATTACCATGAGCAAATCAGAGTGTTTAAAGCA
GGCTTTTAAGCTTGAATGTGTTAGCATGGAATAATGAAATATGACTTTAGTCCTATTTTCGTTGGTTTAGGTACTTC
AGTAATGATGAATAGAAACGGTTAGGGGCATTTGTATTTGGTCGCTAGAGGTGAAATTCTTGGATTGACCGAAGACA
AACTACTGCGAAAGCATTTGACCCGGGACGTTTTCATTGATCAAGGTCTAAAGTTAAGGGATCGAAGACGATTAGAT
ACCGTCGTAGTCTTAACCACAAACTATGCCGACTAGAGATTGGGCGTGTTTATTATGACTCGCTCAGCATCTTAGCG
AAAGTAAAGTTTTTGGGTTCTGGGGGGAGTATGGGACGCAAGGCTGAAACTTAAAGGAATTGACGGAAGGGCACCAC
CAGGAGTGGAGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACATAGTAAGGATTGACAGA
TTGAAAGCTCTTTCTAGATTCTATGGGTGGTGGTGCATGGCCGTTCTTAGTTCGTGGAGTGATTTGTCTGGTTAATT
CCGATAACGAACGAGACCTTATTCTGCTAATTAGACAGGCCAACTCTTTCGGGTTGGTCTATATTATATATTTCACT
GGCTTCTTAGAGAGACTATCGGCTTCAAGCCGAAGGAAGTTTTAGGCAATAACAGGTCTGTGATGC
Embodiment 2：Application of the bacterial strain of the present invention in glutinous rice wine Mashing process
Precise 1Kg content of starch is more than 70% glutinous rice, by cleaning, steeped overnight, boiling 25min, after cooling,
The water of equal quality is added, 1Kg is weighed after mixing；The strain for taking 1 pipe ultralow temperature glycerine freezing pipe to preserve, puts room-temperature dissolution, falls
Enter in culture medium, by stirring, beat nest, ferment 65h under the conditions of 30 DEG C, by 4 layers of filtered through gauze, collect fermentation liquid
793mL, mash total sugar content is 413g/L, glucose 196g/L.
Finally it should be noted that：The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention,
Although the present invention is described in detail with reference to the foregoing embodiments, for a person skilled in the art, it still may be used
To be modified to the technical scheme described in foregoing embodiments, or to which part technical characteristic progress equivalent,
Within the spirit and principles of the invention, any modifications, equivalent substitutions and improvements made etc., should be included in the present invention's
Within protection domain.
Claims (6)
1. one plant of rhizopus stolonifer bacterium, is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation is compiled
Number it is CGMCC NO.13767, Classification And Nomenclature is：Rhizopus stolonifer Cy3, Rhizopus stoloniferCy3.
2. one plant of rhizopus stolonifer and its application, it is characterised in that：The rhizopus stolonifer bacterial strain is screened from glutinous rice wine distiller's yeast and obtained, and is used
In the Mashing process of glutinous rice wine.
3. as claimed in claim 2, the distiller's yeast is tradition glutinous rice wine solid distiller's yeast, carried by Dragon Palace liquid Wine Co., Ltd of Hunan Province
For.
4. the screening technique of the bacterial strain described in claim 2, it is characterized in that：
A, accurately weighs glutinous rice wine solid distiller's yeast 1.0g, is put into sterile mortar, adds 10mL sterilized waters and is ground, wears into paste
Shape, then adds 40mL sterilized waters and is stirred, obtain distiller's yeast suspension, with the ring of oese picking one in isolation medium, with flat
Plate rubbing method is applied in sterile isolation medium, 30 DEG C of culture 15h；Aseptically, with the maximum mould of oese picking
In bacterial strain, 25mL sterile test tubes of the access equipped with 20mL sterilized waters, dilution is stirred, with the ring of oese picking one in single bacterium colony
Screening and culturing medium, is applied in sterile single bacterium colony screening and culturing medium with spread plate, 30 DEG C of culture 36h；Aseptically, use
In the maximum fungal strain of oese picking, 25mL sterile test tubes of the access equipped with 20mL sterilized waters, dilution is stirred, is drawn
1mL is in 1L seed culture mediums, 30 DEG C, under the conditions of 150rpm, cultivates 25h, standby as seed liquor, uses -80 DEG C of ultralow temperature
Glycerine freezing pipe preserving process preservation seed liquor.
Sterile isolation medium described in b, claim a, it is characterised in that nutrient media components is：Sticky rice juice 300mL, chlorine are mould
Plain 0.1g/L, agar 15.0g/L；Sterilizing methods are：Accurately weigh culture medium each component, heating for dissolving in 700ml distilled water,
PH to 6.0 is adjusted with 0.1mol/L Hcl and NaOH solution, triangular flask is dispensed, 121 DEG C of autoclavings 30 minutes take out, sterile
Under the conditions of cooled down, be down flat plate.
C, the single bacterium colony screening and culturing medium described in claim a, it is characterised in that nutrient media components is：Sticky rice juice 300mL, chlorine
Mycin 0.1g/L, agar 15.0g/L, penicillin 80U/mL；Sterilizing methods are：Accurate weighing culture medium each component, heating for dissolving
In 700ml distilled water, pH to 6.0 is adjusted with 0.1mol/L Hcl and NaOH solution, triangular flask, 121 DEG C of autoclavings 30 is dispensed
Minute, take out, aseptically cooled down, be down flat plate.
D, the seed culture medium that claim a is told, it is characterised in that nutrient media components is：Sticky rice juice 300mL, adds 700ml
Distilled water, pH to 6.0 is adjusted with 0.1mol/L Hcl and NaOH solution.
The sticky rice juice of e, claim b, c, d requirement, it is characterized in that:Content of starch is more than 70% glutinous rice 300g, by cleaning,
Add water 1000mL, boiling 40min, after cooling, 4 layers of filtered through gauze, collects the sticky rice juice of filtering.
5. the bacterial strain described in claim 2 is used for the Mashing process that glutinous rice wine is produced, it is characterised in that：Take 1 pipe ultralow temperature sweet
The strain that oily freezing pipe is preserved, puts room-temperature dissolution, pours into 1Kg fermentation mediums, by stirring, beats nest, is issued in 30 DEG C of conditions
Ferment 65h, by 4 layers of filtered through gauze, surveys the total pol of mash.
6. the fermentation medium described in claim 5, it is characterized in that：Content of starch is more than 70% glutinous rice, passes through cleaning, dipping
Overnight, boiling 25min, after cooling, adds the water of equal quality, stand-by.
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| CN111394258A (en) * | 2020-04-02 | 2020-07-10 | 浙江工业大学 | Rhizopus stolonifer F L-3 and application thereof in extracting pachyman |
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| CN111394258A (en) * | 2020-04-02 | 2020-07-10 | 浙江工业大学 | Rhizopus stolonifer F L-3 and application thereof in extracting pachyman |
| CN111394258B (en) * | 2020-04-02 | 2022-03-18 | 浙江工业大学 | Rhizopus stolonifer FL-3 and application thereof in extracting pachyman |
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