CN101781631A - Lactate lowering bacteria and application thereof in white spirit production technology - Google Patents
Lactate lowering bacteria and application thereof in white spirit production technology Download PDFInfo
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- CN101781631A CN101781631A CN 200910273366 CN200910273366A CN101781631A CN 101781631 A CN101781631 A CN 101781631A CN 200910273366 CN200910273366 CN 200910273366 CN 200910273366 A CN200910273366 A CN 200910273366A CN 101781631 A CN101781631 A CN 101781631A
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- bacillus licheniformis
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Abstract
The invention relates to a lactate lowering bacteria and an application thereof in white spirit production technology. The strain is Bacillus licheniformis 331 separated from strong aromatic high-temperature yeast for making hard liquor and utilizes the characteristics that sugar does not produce lactic acid and can favorably grow under the acid environment with pH of 2.5-3.5; the strain is reserved in the China Center for Type Culture Collection (CCTCC) in Wuhan university at Luojia Mountain in Wuchang city, Hubei province on 10, Dec, 2009, and the reservation number is as follows: CCTCC NO: M 209297. The production flow of the invention does not have strict limitation on the original technology of a user, yeast powder for making hard liquor and lactate lowering bacteria are simultaneously added into fermented grains before entering a pit; the mixture is fermented in a pool to evaporate white spirit after being mature, and thus ethyl lactate content in the white spirit of yeast for making hard liquor can be controlled. The lactate lowering bacteria strain has special fermentation characteristics, can obviously lower ethyl lactate content, convert ethyl into ethanol by conversion and improve the yield of white spirit. The white spirit produced by the invention has extremely delicate flavour, soft and sweet fragrance and clean aftertaste, and the quality and yield of white sprit are greatly improved.
Description
Technical field
The present invention relates to a kind of lactate lowering bacteria and be applied in production technique in the liquor, belong to biological chemical field.
Background technology
The ratio of ethyl lactate in several liquor of China and main body fragrant component (ethyl hexanoate or ethyl acetate) is all between 0.3～0.8, and this ratio of high quality liquor is also many between 0.50～0.95.But now ubiquity the too high problem of content of ethyl lactate in the liquor in China's liquor industry, even the ethyl lactate of the product that has surpasses the several times of main body fragrant component, thereby makes serious disproportion.Just in this case, reduce the content of ethyl lactate in the liquor or coordinate it and the ratio of main body fragrant, become the problem that vast liquor-making enterprises presses for solution.
There is the method for content of ethyl lactate in several reduction liquor by solid fermentation in present domestic liquor manufacturer, wherein a kind of is by adding inhibitor or coenzyme during the fermentation, influence the TCA circulation as fumaric acid (as: CN93105171.1 method that reduces content of ethyl lactate in the " Daqu " white spirit) and flavin adenine dinucleotide (FAD), the growth of lactic acid bacteria inhibiting, reduce the concentration of substrate of ethyl lactateization, thereby achieve the goal.But this method also easily suppresses simultaneously the growth of other bacterial classifications, the carrying out and the result of influence fermentation at lactic acid bacteria inhibiting.Because above-mentioned several materials of mentioning are the essential somatomedins of all microorganism growth, must disturb other microbial growth.
Another kind method is by adopting fermentation of reflux liquor to reduce content of ethyl lactate, pluck wine concentration but this method needs strictness to control, and consume a large amount of energy, prolonged fermentation period, restricted the performance of enterprises' production capacity, influencing economic benefit of enterprises.Be to form by a kind of method again, influence its metabolism and meta-bolites component proportions, and then achieve the goal by the multiple microbe colony that changes in the fermenting process Zhong Jiao pond.But kind of microorganism and ratio are difficult to control in this method fermenting process, and the wine taste and the composition of production have nothing in common with each other, and cause the quality instability of wine, and production process is numerous and diverse, expends lot of manpower and material resources, are difficult between the operations coordinate to take into account.
Summary of the invention
The object of the present invention is to provide a kind of lactate lowering bacteria and be applied in production technique in the liquor, the present invention has overcome the open defect of above-mentioned several common methods, and this lactate lowering bacteria utilizes not lactic acid producing of carbohydrate, can be under sour environment pH2.5-3.5 well-grown.The present invention is applicable to all white spirit by solid state method fermentative production, and simple to operate, effect is obvious, does not change original Production Flow Chart, and the quality of wine and local flavor are all stablized.Not only economy but also environmental protection reduce enterprise's spending, increase economic efficiency.
Technical scheme of the present invention is: a kind of lactate lowering bacteria, it is characterized in that: this bacterial strain is the Bacillus licheniformis Bacillus licheniformis 331 that a strain is located away from the Luzhou-flavor high-temperature daqu, this lactate lowering bacteria utilizes not lactic acid producing of carbohydrate, can be under sour environment pH2.5-3.5 well-grown; Be preserved in Wuhan, Hubei loujia hill belongs Wuhan University China typical culture collection center on December 8th, 2009, preserving number is: CCTCC NO:M 209297.
Aforesaid lactate lowering bacteria, it is characterized in that: the enlarged culturing technology of this bacterial strain is: with Chinese sorghum, rice, glutinous rice, wheat, corn is crushed to the 40-80 order respectively, by weight 25-38%, 20-26%, 10-20%, 10-20%, 5-10% is mixed into the grain powder, every liter of substratum contains grain powder 50-100g, prepared culture medium is contained in the triangular flask, with gauze wrapping bottleneck, sterilization, cooling, the inoculation amount is a transfering loop bacterium, 30-35 ℃ of isoperibol cultivated, the centre is shaken 1-3 time, cultivates 2-3 days, obtains Bacillus licheniformis Bacillus licheniformis 331 seeds; The amplification of follow-up lichens bacillus licheniformis 331 amounts is cultivated with above-mentioned same substratum and method, and the inoculation amount is the 5-10% of culture volume, finally obtains every milliliter of lactate lowering bacteria liquid and contains 1-2 * 10
9The lactate lowering bacteria of individual quantity.
Aforesaid lactate lowering bacteria, it is characterized in that: the culture process of this bacterial strain is: Chinese sorghum, rice, glutinous rice, wheat, corn are crushed to 80 orders respectively, 36%, 22%, 18%, 16%, 8% be mixed into the grain powder by weight, every liter of substratum contains grain powder 100g; Prepared culture medium is contained in the triangular flask, with gauze wrapping bottleneck, and sterilization, cooling, the inoculation amount is a transfering loop bacterium, and 35 ℃ of isoperibols are cultivated, and the centre is shaken 2 times, cultivates 3 days; Obtain Bacillus licheniformis Bacillus licheniformis 331 seeds; The amplification of follow-up lichens bacillus licheniformis 331 amounts is cultivated with above-mentioned same substratum and method, and the inoculation amount is 10% of a culture volume, finally obtains every milliliter of lactate lowering bacteria liquid and contains 2 * 10
9The lactate lowering bacteria of individual quantity.
A kind of lactate lowering bacteria is applied in the production technique in the liquor, it is characterized in that: add in the grain unstrained spirits before going into the cellar for storing things in the koji powder, be pressed into cellar for storing things grain powder ratio and add lactate lowering bacteria liquid, be that every 1000Kg goes into to store the grain powder and adds 1-100L lactate lowering bacteria liquid as required, steam wine after the pit entry fermentation maturation, can control content of ethyl lactate in the " Daqu " white spirit; This lactate lowering bacteria is the Bacillus licheniformis 331 that a strain is located away from the Luzhou-flavor high-temperature daqu, is preserved in Wuhan, Hubei loujia hill belongs Wuhan University China typical culture collection center on December 10th, 2009, and preserving number is: CCTCC NO:M 209297.
The invention has the beneficial effects as follows: the present invention adopts specific lactate lowering bacteria (utilizing the bacterium of lactic acid) to participate in fermentation, can significantly reduce content of ethyl lactate, promotes the raising of ethyl hexanoate content.This lactate lowering bacteria utilizes not lactic acid producing of carbohydrate, can be under sour environment pH2.5-3.5 well-grown; This lactate lowering bacteria is picked-up lactic acid and other nutritive substance from the thalline outside atmosphere just, can directly not influence other microorganism; The present invention comprises the production technique that adopts this lactate lowering bacteria, and making lactate lowering bacteria under anaerobic transform lactic acid is the alcoholic acid primary product, can not bring other detrimentally affect to the product local flavor.Strain culturing process and technology are all simple and reliable, and operation the wine factor of merit to improving easily, shorten fermentation period, and actual application value has reduced production costs.
Embodiment
The present invention is further illustrated below in conjunction with embodiment.
The lactate lowering bacteria bacterial strain that the embodiment of the invention relates to is the Bacillus licheniformis 331 that a strain is located away from the Luzhou-flavor high-temperature daqu, and isolation medium (Lu substratum) contains 5g lactic acid for every liter, 2g yeast extract paste, 2g (NH
4)
2SO
2, 14.3gNa
2HPO
412H
2O, 3gKH
2PO
4, 0.28mgMnSO
4H
2O, 0.3mgFeSO
47H
2O, 0.06mgMgSO
47H
2O, 1mgCaCl
2, 0.05mgCuSO
4, 0.05mgZnSO
4And 0.05mgH
3BO
3Get Daqu sample 1g, add sterilized water 10ml, vibration 1min, the transfering loop liquid that takes a morsel is rule every sample 4 wares on the agar isolation medium.Under vacuum condition, cultivate 2-3d for 35 ℃.Obtain single bacterium colony.This bacterial classification purifying on flat board is defined as being used for Physiology and biochemistry evaluation and performance test behind the single culture.36 ℃ of this bacterium optimum growth temperatures are utilized multiple substrate growth such as glucose, fructose, sucrose, lactic acid, and are not produced lactic acid when the static cultivation of liquid.Well-grown under the sour environment (pH.2.5-3.5) of solid medium.Under anaerobic environment, lactic acid be can effectively transform, ethanol, CO generated
2With small amount of acetic acid be main product, be accredited as Bacillus licheniformis (Bacilluslicheniformis) at the VITEK microbial identification system, be preserved in Wuhan, Hubei loujia hill belongs Wuhan University China typical culture collection center on December 10th, 2009, preserving number is: CCTCC NO:M 209297.
Embodiment 1: utilize the growth experiment of lactic acid for matrix: dress 10mlLu liquid nutrient medium inoculation one ring Bacillus licheniformis 331 bacterial classifications in 18 * 180mm glass test tube, 35 ℃ of thermostat containers leave standstill to be cultivated 2 days, get nutrient solution, use efficient liquid phase chromatographic analysis, lactate level reduces by 99.5% than blank, produces 1.01% ethanol, 0.21% acetate simultaneously.
Embodiment 2: the glucose and xylose mixed carbon source leaves standstill liquid fermenting: this substratum is to replace the lactic acid configuration to form with 0.5% glucose and 0.5% wood sugar in the Lu substratum, adorns 10ml in 18 * 180mm glass test tube, inoculation one ring, and 35 ℃ leave standstill cultivation 6d.Get nutrient solution, use efficient liquid phase chromatographic analysis, lactate level remains on the blank level, and promptly 331 bacterium do not have lactic acid to produce.Lactate lowering bacteria 331 does not produce lactic acid when carbohydrate is matrix.
The lactate lowering bacteria enlarged culturing technology that the embodiment of the invention relates to is: with Chinese sorghum, rice, glutinous rice, wheat, corn is crushed to the 40-80 order respectively, by weight 25-38%, 20-26%, 10-20%, 10-20%, 5-10% is mixed into the grain powder, every liter of substratum contains grain powder 50-100g, prepared culture medium is contained in the triangular flask, adorn 2/3 volume, with 4 layers of gauze wrapping bottleneck, sterilization, cooling, the inoculation amount is a transfering loop bacterium, 30-35 ℃ of isoperibol cultivated, the centre is shaken 1-3 time, cultivates 2-3 days, obtains Bacillus licheniformis Bacillus licheniformis 331 seeds; The amplification of follow-up lichens bacillus licheniformis 331 amounts is cultivated with above-mentioned same substratum and method, and the inoculation amount is the 5-10% of culture volume, finally obtains every milliliter of lactate lowering bacteria liquid and contains 1-2 * 10
9The lactate lowering bacteria of individual quantity.
The lactate lowering bacteria culture process that the embodiment of the invention relates to is preferably: the culture process of this bacterial strain is: Chinese sorghum, rice, glutinous rice, wheat, corn are crushed to 80 orders respectively, 36%, 22%, 18%, 16%, 8% be mixed into the grain powder by weight, every liter of substratum contains grain powder 100g; Prepared culture medium is contained in the triangular flask, adorns 2/3 volume, with 4 layers of gauze wrapping bottleneck, and sterilization, cooling, the inoculation amount is a transfering loop bacterium, and 35 ℃ of isoperibols are cultivated, and the centre is shaken 2 times, cultivates 3 days; Obtain Bacillus licheniformis Bacillus licheniformis 331 seeds; The amplification of follow-up lichens bacillus licheniformis 331 amounts is cultivated with above-mentioned same substratum and method, and the inoculation amount is 10% of a culture volume, finally obtains every milliliter of lactate lowering bacteria liquid and contains 2 * 10
9The lactate lowering bacteria of individual quantity.
The embodiment of the invention adopts specific lactate lowering bacteria to strengthen that microorganism makes lactic acid content controlled to the effect that lactic acid utilizes in the liquor solid fermentation.The embodiment of the invention is utilized the autonomous isolating lactate lowering bacteria bacterial classification with property of a strain, and in the process of fermentation, the bacterium liquid that it is an amount of directly adds wine unstrained spirits pit entry fermentation, can reduce content of ethyl lactate.This lactate lowering bacteria strain fermentation character uniqueness, it is obvious to reduce the content of ethyl lactate effect, and lactic acid can be become ethanol by metabolic conversion, has increased the productive rate of liquor.Particularly when carbohydrate is matrix, do not produce the composition of lactic acid and other conspicuous level, do not bring any peculiar smell sense, aroma is outstanding, and is sweet soft, and pleasant impression is refreshing clean, improved the quality and the output of wine greatly.
The embodiment of the invention relates to Production Flow Chart does not have strict restriction to the original technology of user, as long as by the ordinary production flow process, lactate lowering bacteria of the present invention and Daqu are added the grain unstrained spirits (20 ℃-30 ℃) of spreading for cooling simultaneously, pile up more than two hours, pit entry fermentation gets final product then.Because it is less relatively to add lactate lowering bacteria quantity, to going into to store the acidity value influence very little (＜10%) of grain unstrained spirits, can be by former technology controlling and process acidity.Adjust the own newborn of wine for reaching,, adjust adding lactate lowering bacteria quantity according to pond, correspondence cellar for storing things situation than purpose.Ethyl lactate and ethyl hexanoate ratio are reached 1.5: 1 situation, press 1L/100Kg grain powder and add lactate lowering bacteria; The situation than 1.3: 1 to ethyl lactate and ethyl hexanoate, adding reduces by half.
Embodiment 3:
1000Kg grain powder adding mother is poor, ripe chaff is joined poor, steaming material, and spreading for cooling is got 200Kg koji powder, 10.00L concentration 10 after 20-30 ℃
9The lactate lowering bacteria of the present invention 331 bacterium liquid of individual/ml are evenly sneaked into the grain unstrained spirits, and pit entry fermentation steamed wine after 40 days.Go out wine analytical results such as table 1.Ethyl lactate reduces by 29%, newborn own eurythmy, and the yield of liquor increases.High-grade-goods rate improves 6%.Go out sweet ice-cold, the tail of aromatic strongly fragrant, the sweet-smelling in the wine cellar length of distinguishing the flavor of only.
Table 1 embodiment 3 lactate lowering bacterias reduce aromatic Chinese spirit content of ethyl lactate effect
| Type | Content of ethyl lactate | Ethyl hexanoate content | The own ratio of breast | Sense organ |
| The contrast cellar for storing things | ??3105mg/L | ??2070mg/L | ??1.5∶1 | Inharmonious |
[0023]
| Lactate lowering bacteria is handled the cellar for storing things | ??2204mg/L | ??2588mg/L | ??0.85∶1 | Coordinate |
Embodiment 4: 1000Kg grain powder adding mother is poor, ripe chaff is joined poor, steaming material, and spreading for cooling is got 200Kg koji powder, 5.00L concentration 10 after 20-30 ℃
9The lactate lowering bacteria of the present invention 331 bacterium liquid of individual/ml are evenly sneaked into the grain unstrained spirits, and pit entry fermentation steamed wine after 40 days.Ethyl lactate reduces by 12%, newborn own eurythmy, and the yield of liquor increases.High-grade-goods rate improves 6%.Local flavor is better than not adding the contrast of lactate lowering bacteria.
Embodiment 5:1000Kg grain powder adding mother is poor, ripe chaff is joined poor, steaming material, and spreading for cooling is got 200Kg koji powder, adding lactate lowering bacteria 331 bacterium liquid 8.00L after 20-30 ℃, evenly sneak into atomizer and store the wine unstrained spirits, and normal the fermentation got unstrained spirits and steamed wine after 40 days.Content of ethyl lactate reduces by 20.1%, ethyl hexanoate content 2521mg/L, and quality percentage improves 5%.Local flavor obviously is better than contrast.
From the foregoing description 3,4,5 as can be seen, lactate lowering bacteria 331 utilizes not lactic acid producing of carbohydrate, can be under sour environment pH2.5-3.5 well-grown; And obviously reduced the content of ethyl lactate in the liquor.
Claims (4)
1. lactate lowering bacteria, it is characterized in that: this bacterial strain is the Bacillus licheniformis Bacillus licheniformis 331 that a strain is located away from the Luzhou-flavor high-temperature daqu, and this lactate lowering bacteria utilizes not lactic acid producing of carbohydrate, can be under sour environment pH2.5-3.5 well-grown; Be preserved in Wuhan, Hubei loujia hill belongs Wuhan University China typical culture collection center on December 10th, 2009, preserving number is: CCTCC NO:M 209297.
2. lactate lowering bacteria according to claim 1, it is characterized in that: the enlarged culturing technology of this bacterial strain is: Chinese sorghum, rice, glutinous rice, wheat, corn are crushed to the 40-80 order respectively, be mixed into the grain powder by weight 25-38%, 20-26%, 10-20%, 10-20%, 5-10%, every liter of substratum contains grain powder 50-100g, prepared culture medium is contained in the triangular flask, with gauze wrapping bottleneck, sterilization, cooling, the inoculation amount is a transfering loop bacterium; 30-35 ℃ of isoperibol cultivated, and the centre is shaken 1-3 time, cultivates 2-3 days, obtains Bacillus licheniformis Bacilluslicheniformis 331 seeds;
The amplification of follow-up lichens bacillus licheniformis 331 amounts is cultivated with above-mentioned same substratum and method, and the inoculation amount is the 5-10% of culture volume, finally obtains every milliliter of lactate lowering bacteria liquid and contains 1-2 * 10
9The lactate lowering bacteria of individual quantity.
3. lactate lowering bacteria according to claim 1 and 2, it is characterized in that: the culture process of this bacterial strain is: Chinese sorghum, rice, glutinous rice, wheat, corn are crushed to 80 orders respectively, 36%, 22%, 18%, 16%, 8% be mixed into the grain powder by weight, every liter of substratum contains grain powder 100g; Prepared culture medium is contained in the triangular flask, with gauze wrapping bottleneck, and sterilization, cooling, the inoculation amount is a transfering loop bacterium; 35 ℃ of isoperibols are cultivated, and the centre is shaken 2 times, cultivates 3 days; Obtain Bacillus licheniformis Bacillus licheniformis 331 seeds;
The amplification of follow-up lichens bacillus licheniformis 331 amounts is cultivated with above-mentioned same substratum and method, and the inoculation amount is 10% of a culture volume, finally obtains every milliliter of lactate lowering bacteria liquid and contains 2 * 10
9The lactate lowering bacteria of individual quantity.
4. a lactate lowering bacteria is applied in the production technique in the liquor, it is characterized in that: add in the grain unstrained spirits before going into the cellar for storing things in the koji powder, be pressed into cellar for storing things grain powder ratio and add lactate lowering bacteria liquid, be that every 1000Kg goes into to store the grain powder and adds 1-100L lactate lowering bacteria liquid as required, steam wine after the pit entry fermentation maturation, can control content of ethyl lactate in the " Daqu " white spirit; Lactate lowering bacteria is the Bacillus licheniformis 331 that a strain is located away from the Luzhou-flavor high-temperature daqu, is preserved in Wuhan, Hubei loujia hill belongs Wuhan University China typical culture collection center on December 10th, 2009, and preserving number is: CCTCC NO:M 209297.
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| CN 200910273366 CN101781631B (en) | 2009-12-23 | 2009-12-23 | Lactate lowering bacteria and culture process and application thereof in white spirit production technology |
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| CN101781631B CN101781631B (en) | 2013-10-09 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102140427A (en) * | 2010-09-29 | 2011-08-03 | 湖北白云边酒业股份有限公司 | Method for preparing intensified daqu applied to nongjiang-flavor Chinese spirits |
| CN113186116A (en) * | 2021-02-08 | 2021-07-30 | 茅台学院 | Non-decarboxylation lechlenibacter NJ22 with lactic acid as carbon source and application thereof |
| CN115044479A (en) * | 2022-04-13 | 2022-09-13 | 茅台学院 | Geotrichum candidum strain with high lactic acid tolerance and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009009391A2 (en) * | 2007-07-06 | 2009-01-15 | Ls9, Inc. | Systems and methods for the production of fatty esters |
| CN101263870B (en) * | 2008-03-28 | 2010-11-10 | 宜兴市天石饲料有限公司 | Highly effective biological acidulant and preparation thereof |
-
2009
- 2009-12-23 CN CN 200910273366 patent/CN101781631B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102140427A (en) * | 2010-09-29 | 2011-08-03 | 湖北白云边酒业股份有限公司 | Method for preparing intensified daqu applied to nongjiang-flavor Chinese spirits |
| CN102140427B (en) * | 2010-09-29 | 2012-10-03 | 湖北白云边酒业股份有限公司 | Method for preparing intensified daqu applied to nongjiang-flavor Chinese spirits |
| CN113186116A (en) * | 2021-02-08 | 2021-07-30 | 茅台学院 | Non-decarboxylation lechlenibacter NJ22 with lactic acid as carbon source and application thereof |
| CN113186116B (en) * | 2021-02-08 | 2022-09-23 | 茅台学院 | Non-decarboxylation lechlenibacter NJ22 with lactic acid as carbon source and application thereof |
| CN115044479A (en) * | 2022-04-13 | 2022-09-13 | 茅台学院 | Geotrichum candidum strain with high lactic acid tolerance and application thereof |
| CN115044479B (en) * | 2022-04-13 | 2024-03-29 | 茅台学院 | Geotrichum candidum strain with high lactic acid tolerance and application thereof |
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