CN105713861A - Method for preparing novel bacteria culture medium from Maotai-flavor liquor vinasse - Google Patents
Method for preparing novel bacteria culture medium from Maotai-flavor liquor vinasse Download PDFInfo
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- CN105713861A CN105713861A CN201610149407.XA CN201610149407A CN105713861A CN 105713861 A CN105713861 A CN 105713861A CN 201610149407 A CN201610149407 A CN 201610149407A CN 105713861 A CN105713861 A CN 105713861A
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- maotai
- enzymolysis
- culture medium
- flavor liquor
- peptone
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention belongs to the field of preparation of biological culture media, and relates to a preparation method of a novel bacteria culture medium. The preparation method comprises the steps that 1, water is added to fresh Maotai-flavor liquor vinasse for enzymolysis, and a filtrate is obtained after filtration; 2, the filtrate is concentrated to be paste or continues to be dried; 3, the concentrate replaces peptone in the bacteria culture medium with different ratios, and typical bacteria culture verification is carried out. The method has the advantages that the brewing by-product liquor vinasse is used as the raw material for enzymolysis, and then the product replaces the peptone with different ratios to be used as a nitrogen source to culture bacteria, peptone is saved, the liquor vinasse is prevented from polluting environment, and meanwhile the using value of the liquor vinasse is improved. The concentrate replaces the peptone with different ratios to be used as the nitrogen source, tested bacteria are cultured for 18-24 h, and the OD value of the concentrate is generally higher than that of the peptone used as the nitrogen source. The preparation method has the good application prospect in the production field of microorganism culture medium nitrogen sources.
Description
Technical field
The invention belongs to the preparation field of microbiological culture media, particularly to a kind of method preparing unicellular microorganism culture medium with Maotai-flavor liquor grain.
Technical background
Distillers ' grains is Chinese liquor solid fermentation and the leftover bits and pieces of solid state distillation traditional fermentation technique, wherein remaining many fully utilized nutritional labelings that fails, particularly rich in the somatomedin needed for growth of microorganism, and Maotai-flavor liquor is lost grain and is caused its crude protein to lose grain apparently higher than other aromatic white spirit because of Maotai-flavor liquor solid fermentation uniqueness production technology, through measuring, crude protein content may be up to 23%.Therefore the present invention considers that using primary raw material is that Maotai-flavor liquor loses grain.The utilization of current distillers ' grains mainly has production chemical products；Cultivate edible fungi；Making vinegar and produce feedstuff etc., its utilization rate and added value still await further raising, and spirit distiller grain improper storage also easy contaminated environment.
China is a conventional brew big country, and distiller grains resource is very abundant, but the deep exploitation rate of distiller grains is non-normally low, spirit distiller grain, and especially the potentiality of the recycling that Maotai-flavor liquor loses grain are also very big.The main restricting factor of the restriction distiller grains utilization of resources has following two aspect: one is fresh grain stillage acidity big (pH is less than 4), and moisture high (55% ~ 60%) is easily putrid and deteriorated, is not easy to transport and storage.Two is blended substantial amounts of rice husk in brewing process, and in dry, rice husk accounts for about the 36% of dry matter weight, and volume accounts for about 50%, and crude fiber content is up to more than 18%.
Summary of the invention
It is an object of the invention to provide a kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain.The method turns waste into wealth, and technique is simple, cost is low, it is easy to operation, is suitable for large-scale production.
A kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain of the present invention, including:
(1) fresh Maotai-flavor liquor grain directly being added water and regulate pH with solid caustic soda, under preference temperature, substep carries out enzymolysis；Fresh Maotai-flavor liquor grain is 1g:8 ~ 12ml with the mass volume ratio of water, fully mixes, carries out one-level enzymolysis, enzymolysis complete after in 90 ~ 95 DEG C, 10 ~ 15min enzyme denaturing；Then carry out two grades of enzymolysis, enzymolysis complete after in 90 ~ 95 DEG C, 10 ~ 15min enzyme denaturing, be filtrated to get hydrolyzed solution；
(2) said hydrolyzed liquid is concentrated into paste, or continues to be dried to solid；
(3) said hydrolyzed product equivalent substitution or part are substituted the peptone in antibacterial minimal medium, be configured to culture medium, after inoculation, carry out antibacterial culturing, after cultivating 18 ~ 24h, measure the Biomass (OD value) of thalline.
In step (1), enzyme includes alkaline protease (1 × 105And papain (1 × 10 U/g)5U/g)；Its addition is followed successively by 500 ~ 2500U/g distiller's dried grain, 250 ~ 2000U/g distiller's dried grain enzymolysis order is followed successively by alkaline protease and papain or papain and alkaline protease.
Solid caustic soda in step (1) is sodium hydroxide, potassium hydroxide or calcium hydroxide.
Step (1) regulates pH and includes alkaline protease enzymolysis pH7.0 ~ 13.0, papain enzymolysis pH6.0 ~ 7.0.
In step (1), preference temperature is alkaline protease 40 ~ 55 DEG C, papain 50 ~ 55 DEG C.
In step (1), enzymolysis time is alkaline protease enzymolysis 2 ~ 4h, papain 1 ~ 3h.
In step (2) described in step (2), hydrolyzed solution is concentrated into paste condition is cryoconcentration, and temperature is less than 80 DEG C, and being concentrated into moisture is 40 ~ 50%, obtains paste；Or continuing dry, baking temperature is 70 ~ 90 DEG C, and vacuum is 0.008 ~ 0.012MPa, is dried to water content lower than 5%.
In step (3), in hydrolyzate substitutive medium, the ratio of peptone is followed successively by 0%, 50%, 100%.
In step (3), minimal medium is beef-protein medium: Carnis Bovis seu Bubali cream 3.0g, peptone 10.0g, NaCl20.0g, water 1000.0ml, pH7.0 ~ 7.2.
Antibacterial in step (3) is escherichia coli, staphylococcus aureus, bacillus subtilis and bacillus thuringiensis.
In step (3), inoculum concentration is 0.5% ~ 3%.
The present invention utilizes Liquor-making industry leftover bits and pieces Maotai-flavor liquor grain to prepare Novel bacterial culture medium for raw material, peptone is substituted with the enzymatic hydrolysate of distiller grains, both the cost using peptone as culture media nitrogen source had been reduced, prevent again the distiller grains pollution to environment, improve the use value of distiller grains, turn waste into wealth.The nitrogenous source that the present invention produces accounts for the 48 ~ 61% of raw material total nitrogen, dried nitrogen content 4.0% ~ 6.5%.Through waiting mass substitution and part to substitute after nitrogenous source peptone, wait mass substitution or part substitute peptone after the Biomass of thalline add 1.5 ~ 3.2 times.Maotai-flavor distiller grains are possible not only to substitute peptone as the nitrogenous source in antibacterial culturing, and advantageously reduce culture medium cost, pollute for solution Chinese liquor using solid distillers ' grains again and provide an approach more, therefore the present invention and method thereof have potential good promotional value.
Below in conjunction with embodiment, the present invention will be further described.
Accompanying drawing explanation
Fig. 1 replaces or partly replaces the peptone in culture medium to cultivate the result of antibacterial after Maotai-flavor liquor grain enzymolysis.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are merely to illustrate the present invention rather than restriction the scope of the present invention.Should be understood that outside secondary that the present invention can be changed or revise by those skilled in the art, and these equivalent form of values fall within the present patent application appended claims limited range equally after having read the content that the present invention lectures.
The preparation of embodiment 1 Maotai-flavor liquor grain hydrolysate comprises the following steps:
(1) fresh Maotai-flavor liquor grain directly being added water and regulate pH with solid caustic soda, under preference temperature, substep carries out enzymolysis；Fresh Maotai-flavor liquor grain is 1g:8 ~ 12ml(example 1g:8ml, 1g:10ml, 1g:12ml with the mass volume ratio of water) fully mix, carry out one-level enzymolysis, enzymolysis complete after in 90 ~ 95 DEG C, 10 ~ 15min enzyme denaturing；Then carry out two grades of enzymolysis, enzymolysis complete after in 90 ~ 95 DEG C, 10 ~ 15min enzyme denaturing, be filtrated to get hydrolyzed solution；
(2) said hydrolyzed liquid is concentrated into paste, or continues to be dried to solid；
In step (1), enzyme includes alkaline protease (1 × 105And papain (1 × 10 U/g)5U/g)；Its addition is followed successively by 500 ~ 2500U/g distiller's dried grain (example 1000U/g, 1500U/g, 2000U/g), 250 ~ 2000U/g distiller's dried grain (example 500U/g, 1000U/g, 2000U/g) enzymolysis order is followed successively by alkaline protease and papain or papain and alkaline protease.
Alkali in step (1) is sodium hydroxide, potassium hydroxide or calcium hydroxide.
Step (1) regulates pH and includes alkaline protease enzymolysis pH7.0 ~ 13.0(example pH7.0, pH9.0, pH11.0), papain enzymolysis pH6.0 ~ 7.0(example pH6.0, pH7.0).
In step (1), preference temperature is alkaline protease 40 ~ 55 DEG C (example 45 DEG C, 50 DEG C, 55 DEG C), papain 50 ~ 55 DEG C (example 50 DEG C, 55 DEG C).
In step (1), enzymolysis time is alkaline protease enzymolysis 2 ~ 4h(example 2h, 3h, 4h), papain 1 ~ 3h(example 1h, 2h, 3h).
In step (2), hydrolyzed solution is concentrated into paste condition is cryoconcentration, and temperature is less than 80 DEG C (examples 60 DEG C, 70 DEG C, 80 DEG C), and being concentrated into moisture is 40 ~ 50%(example 40%, 45%, 50%), obtain paste；Or continue dry, baking temperature be 70 ~ 90 DEG C (example 70 DEG C, 80 DEG C, 90 DEG C), vacuum is 0.008 ~ 0.012MPa(example 0.008MPa, 0.010MPa, 0.012MPa), be dried to water content lower than 5%.
Embodiment 2 Maotai-flavor liquor grain hydrolysate substitutes nitrogenous source peptone and cultivates escherichia coli
Nitrogenous source peptone in the Maotai-flavor liquor grain hydrolysate substitutive medium obtained is arranged process as follows: CK(beef-protein medium) hydrolysate replacement amount 50%, 100% totally 3 process, medium pH is unified is adjusted to 7.2, the bottled 50ml test medium of 250ml triangle, 121 DEG C of sterilizing 20min.Take activated escherichia coli seed culture fluid (30 DEG C, 170r/min cultivate 18h) and be seeded in the triangular flask of sterilizing 30 DEG C, 170r/min according to 1%, after culture fluid being diluted 10 times after cultivating 24h, survey light absorption value.Obtain corresponding light absorption value, result: CK light absorption value is 0.186；50% replacement amount light absorption value is 0.281；100% replacement amount light absorption value is 0.2375.The light absorption value of 50% replacement amount is 1.5 times of CK；The light absorption value of 100% replacement amount is 1.2 times of CK.
Embodiment 3 Maotai-flavor liquor grain hydrolysate substitutes nitrogenous source peptone and cultivates bacillus subtilis
Nitrogenous source peptone in the Maotai-flavor liquor grain hydrolysate substitutive medium obtained is arranged process as follows: CK(beef-protein medium) hydrolysate replacement amount 50%, 100% totally 3 process, medium pH is unified is adjusted to 7.2, the bottled 50ml test medium of 250ml triangle, 121 DEG C of sterilizing 20min.Take activated bacillus subtilis seed culture fluid (30 DEG C, 170r/min cultivate 18h) and be seeded in the triangular flask of sterilizing 30 DEG C, 170r/min according to 1%, after culture fluid being diluted 10 times after cultivating 24h, survey light absorption value.Obtain corresponding light absorption value, result: CK light absorption value is 0.193；50% replacement amount light absorption value is 0.566；100% replacement amount light absorption value is 0.482.The light absorption value of 50% replacement amount is 3.2 times of CK；The light absorption value of 100% replacement amount is 2.5 times of CK.
Aminoacid after the hydrolysis of table 1 Maotai-flavor liquor grain distinct methods forms and content
Claims (9)
1. a method for Novel bacterial culture medium is prepared with Maotai-flavor liquor grain, including:
(1) fresh Maotai-flavor liquor grain directly being added water and regulate pH with solid caustic soda sodium hydroxide, potassium hydroxide or calcium hydroxide, under preference temperature, substep carries out enzymolysis；Fresh Maotai-flavor liquor grain is 1g:8 ~ 12ml with the mass volume ratio of water, fully mixes, carries out one-level enzymolysis, enzymolysis complete after in 90 ~ 95 DEG C, 10 ~ 15min enzyme denaturing；Then carry out two grades of enzymolysis, enzymolysis complete after in 90 ~ 95 DEG C, 10 ~ 15min enzyme denaturing, be filtrated to get hydrolyzed solution；
(2) said hydrolyzed liquid is concentrated into paste, or continues to be dried to solid；
(3) said hydrolyzed product equivalent substitution or part are substituted the peptone in antibacterial minimal medium, be configured to culture medium, after inoculation, carry out antibacterial culturing, after cultivating 18 ~ 24h, measure the Biomass (OD value) of thalline.
2. a kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain according to claim 1, it is characterised in that: in described step (1), enzyme includes alkaline protease (1 × 105And papain (1 × 10 U/g)5U/g)；Its addition is followed successively by 500 ~ 2500U/g distiller's dried grain, 250 ~ 2000U/g distiller's dried grain enzymolysis order is followed successively by alkaline protease and papain or papain and alkaline protease.
3. a kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain according to claim 1, is characterised by: regulating pH in described step (1) and preference temperature includes alkaline protease enzymolysis pH7.0 ~ 13.0, preference temperature is 40 ~ 55 DEG C；Papain enzymolysis pH6.0 ~ 7.0, preference temperature is 50 ~ 55 DEG C.
4. a kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain according to claim 1, is characterised by: in described step (1), enzymolysis time is alkaline protease enzymolysis 2 ~ 4h, papain 1 ~ 3h.
5. a kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain according to claim 1, it is characterised by: in described step (2), hydrolyzed solution is concentrated into paste condition is cryoconcentration, temperature is less than 80 DEG C, and being concentrated into moisture is 40 ~ 50%, obtains paste；Or continuing dry, baking temperature is 70 ~ 90 DEG C, and vacuum is 0.008 ~ 0.012MPa, is dried to water content lower than 5%.
6. a kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain according to claim 1, is characterised by: in described step (3), in hydrolyzate substitutive medium, the ratio of peptone is followed successively by 0%, 50%, 100%.
7. a kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain according to claim 1, is characterised by: in described step (3), minimal medium is beef-protein medium: Carnis Bovis seu Bubali cream 3.0g, peptone 10.0g, NaCl20.0g, water 1000.0ml, pH7.0 ~ 7.2.
8. a kind of method preparing Novel bacterial culture medium with Maotai-flavor liquor grain according to claim 1, it is characterised in that: the antibacterial in described step (3) is escherichia coli, staphylococcus aureus, bacillus subtilis and bacillus thuringiensis.
9. a kind of method preparing bacteria culture media with Maotai-flavor liquor grain according to claim 1, it is characterised in that: in described step (3), inoculum concentration is 0.5% ~ 3%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111518727A (en) * | 2020-05-09 | 2020-08-11 | 李晟雪 | High-strength and high-density fermentation medium for bacillus subtilis and method for preparing microbial inoculum by fermenting bacillus subtilis |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102965305A (en) * | 2012-11-02 | 2013-03-13 | 湖南天之衡酒业有限公司 | Utilization method of white spirit distiller's grains |
| CN102960541A (en) * | 2012-11-30 | 2013-03-13 | 张有聪 | Method for producing environment-friendly biological feed employing distiller grains and glycoprotein |
| CN103074284A (en) * | 2013-01-28 | 2013-05-01 | 四川国凤生物科技有限公司 | Zymophyte liquid, liquid spirit stillage fermented feed and fermentation method of liquid spirit stillage fermented feed |
| CN103689224A (en) * | 2013-12-07 | 2014-04-02 | 路德环境科技股份有限公司 | Zymolytic spirit vinasse high-peptide feed and production method thereof |
| CN104046583A (en) * | 2014-06-19 | 2014-09-17 | 泸州品创科技有限公司 | Bacillus thuringiensis liquid culture medium and preparation method thereof |
| CN105256081A (en) * | 2015-11-06 | 2016-01-20 | 宜宾学院 | Method for preparing xylose by utilizing distiller's grains of Baijiu |
-
2016
- 2016-03-16 CN CN201610149407.XA patent/CN105713861A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102965305A (en) * | 2012-11-02 | 2013-03-13 | 湖南天之衡酒业有限公司 | Utilization method of white spirit distiller's grains |
| CN102960541A (en) * | 2012-11-30 | 2013-03-13 | 张有聪 | Method for producing environment-friendly biological feed employing distiller grains and glycoprotein |
| CN103074284A (en) * | 2013-01-28 | 2013-05-01 | 四川国凤生物科技有限公司 | Zymophyte liquid, liquid spirit stillage fermented feed and fermentation method of liquid spirit stillage fermented feed |
| CN103689224A (en) * | 2013-12-07 | 2014-04-02 | 路德环境科技股份有限公司 | Zymolytic spirit vinasse high-peptide feed and production method thereof |
| CN104046583A (en) * | 2014-06-19 | 2014-09-17 | 泸州品创科技有限公司 | Bacillus thuringiensis liquid culture medium and preparation method thereof |
| CN105256081A (en) * | 2015-11-06 | 2016-01-20 | 宜宾学院 | Method for preparing xylose by utilizing distiller's grains of Baijiu |
Non-Patent Citations (2)
| Title |
|---|
| 邹明鑫等: "利用酱香型白酒丢糟生产微生物饲料添加剂的初步研究", 《酿酒科技》 * |
| 郭志等: "白酒丢糟饲料化的发酵培养基优化研究", 《粮食与饲料工业》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111518727A (en) * | 2020-05-09 | 2020-08-11 | 李晟雪 | High-strength and high-density fermentation medium for bacillus subtilis and method for preparing microbial inoculum by fermenting bacillus subtilis |
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