CN101153260A - Method of preventing and controlling velum in ferment production of liquid condition pouring edible vinegar - Google Patents
Method of preventing and controlling velum in ferment production of liquid condition pouring edible vinegar Download PDFInfo
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- CN101153260A CN101153260A CNA2007100779203A CN200710077920A CN101153260A CN 101153260 A CN101153260 A CN 101153260A CN A2007100779203 A CNA2007100779203 A CN A2007100779203A CN 200710077920 A CN200710077920 A CN 200710077920A CN 101153260 A CN101153260 A CN 101153260A
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Abstract
The invention discloses a velum prevention method in liquid leaching vinegar fermentation production. The method is that the acidic cellulose which is not lower than 0.08 percent of the total mass of the material is added once for all during the acetic acid fermentation phase of the process; the preferable mass of the added acidic cellulose is 0.08 percent of the total mass of the material; when the acidic cellulose is added, no bacteria operation is done to avoid the pollution of the bacteria; the added acidic cellulose is the product sold in the market, and the most suitable enzyme-catalyzed reaction conditions of the cellulose are that: the enzyme-catalyzed reaction time is 30min, pH value is 5.0, the enzyme fluid is diluted to 130 times, and the enzyme activity under 50 DEG C is 1685IU/g. The method of the invention studies the bacterial cellulose from the angle that how to decrease the yield of the bacterial cellulose or the bacterial cellulose is radically eliminated for the first time, and invents the velum prevention method during the acetic acid fermentation; the method of the invention can radically prevent the bacterial cellulose velum and ensure the long-period normal operation of the liquid leaching vinegar fermentation production. The method of the invention is suitable for the enterprises which adopt the liquid leaching vinegar fermentation process.
Description
Technical field
The present invention relates to the preparation of vinegar, the method for preventing and treating of mycoderm in the liquid sprinkling vinegar fermentative production.
Background technology
The liquid sprinkling fermentation method of vinegar is widely used in the world advanced vinegar brewing method, the external raw material that is adopted is a low-concentration ethanol liquid, China is then on the basis of traditional vinegar brewing, brewage as the acetic fermentation raw material after zymamsis with enzyme process rice saccharification liquid, obtained good economic benefits.But, ubiquitous serious problems are after refluxing through sprinkling about 1～March in the liquid sprinkling fermentative Production of China's vinegar at present, Ta Neiyi forms the transparent mycoderm of leather look like fibrous, be commonly called as Hai Yue, this film has stopped up the space between stopping composition, because air and vinegar liquid can not proper flow and must stop fermentation.The liquid sprinkling fermentation vinegar of external particularly Germany is because employing low-concentration ethanol liquid is raw material, even long-term the backflow can not produce membranoid substance yet, its produce product be alcohol vinegar (Shen Zhengxie, section is beautiful. Japanese vinegar (the translation choosing is carried) [J]. China brewages, 1994,1:36-43); And China is raw material with the rice, and the product of production is a rice vinegar.The moiety of mycoderm has data to think Mierocrystalline cellulose, promptly by bacteriogenic Mierocrystalline cellulose; The data that it produces reason has is thought to produce in the production process due to the pollution of film bacterium; Also having data to think that most acetic bacteria all is to produce the film bacterium, is because due to the secular backflow fermentation.The solution of the domestic report of this problem is few, Xie Kaixi, indigo plant are defended stopping composition are carried out orderly arrangement, make stopping composition than more abundant by fermented liquid " spray " in the past, the harsh length of mycoderm just swept away and can not gather (to improve one's methods [J] of the liquid sprinkling fermentation tower of traditional vinegar. Chinese seasonings, 2001,4:9-33); The report of Zhang Chunzhi etc. then thinks, " can adopt a large amount of Shanghai to make growth and breeding that 1.01 acetic acid floras suppress " mycoderm " " (tower vinegar produce in the influence and the control [J] of " Hai Yue ". Dalian Polytechnic College journal, 1997,4 (16): 36-39).Above reported method all can only be taken stopgap measures and can not be effected a permanent cure.
Summary of the invention
The object of the present invention is to provide the method for preventing and treating of mycoderm in the liquid sprinkling vinegar fermentative production, fundamentally prevent the generation of bacteria cellulose film, guarantee that the long period of liquid sprinkling vinegar fermentative production is normally moved.
For the reason of finding out the mycoderm generation and the method for preventing and treating mycoderm, the contriver has at first carried out purebred acetic bacteria and bacteria cellulose in testing laboratory and has produced the separating of bacterium, purifying and cultivation, next has analyzed the main component of mycoderm, last under the prerequisite that does not influence acetic fermentation, find out the method that the control mycoderm produces, and tested out best processing condition.
The concrete grammar that the contriver carries out in testing laboratory comprises following 3 steps:
1. bacteria cellulose produces separation, purifying, the cultivation of bacterium:
Separate: get the stopping composition corn cob in the long film fermentation jar in certain company liquid sprinkling vinegar fermentative production, in the liquid state fermentation substratum, carry out enrichment culture; The enrichment culture liquid of getting the long film of 1ml is diluted to 10 respectively
-1～10
-8, draw back 3 suitable dilution diluent 0.1ml respectively on the solid plate substratum, use aseptic spreading rod spread plate immediately; Be upside down in then in 30 ℃ of constant incubators, cultivate about 6d; Single bacterium colony on the picking flat board inserts slant medium, cultivates 2d for 30 ℃; Slant strains is inserted in the liquid fermentation medium, 30 ℃, 120r/min constant temperature shaking table cultivation 12h leave standstill cultivation afterwards in 30 ℃ of constant incubators again, and whether observe has mycoderm to produce.
Purifying: produce the film slant strains with the transfering loop picking in liquid fermentation medium, dilution is coated with flat board as stated above, is upside down in then and cultivates the single bacterium colony that occurs homogeneous on flat board in 30 ℃ of constant incubators; Picking list bacterium colony connects people's slant medium, cultivates 2d for 30 ℃; Again slant strains is inserted in the liquid fermentation medium, have mycoderm to produce, illustrate that institute's strain separated is the purifying bacterial strain.
Cultivate: choose the new inclined-plane bacterium of cultivating of 1-2 ring and inscribe in the test tube liquid nutrient medium, behind the cultivation 12h, inoculation 2%～10% is in triangular flask liquid nutrient medium or alcohol fermented beer, and 30 ℃ are shaken bottle or leave standstill cultivation, carry out the extraction and the processing of film behind 6～7d.
2. the qualitative examination of mycoderm component:
X-ray diffraction method of testing by qualitative observation Mierocrystalline cellulose method, cellulase hydrolysis method, scanning electron microscopic observation dry film configuration of surface structure method and dry film has carried out qualitative examination to the mycoderm component; The qualitative examination result of several mycoderm components shows that the composition of mycoderm is a Mierocrystalline cellulose, promptly by bacteriogenic Mierocrystalline cellulose.
3. carry out the control test of mycoderm:
With separate, the inclined-plane seed of the acetic bacteria of purifying activates, and insert 50m1 acetic bacteria fermention medium by 6% inoculum size, substratum is from the alcohol fermented beer of rice saccharification liquid in producing, and its alcoholic strength is 6.3% (volume fraction), and every group parallel makees 3 samples; Do the blank except that one group, other group adds commercially available acidic cellulase (buying from Kweiyang Sai Lanbo company) by 0%, 0.02%, 0.05%, 0.08%, 0.1%, 0.3%, 0.5%, 0.8%, the optimum condition of this cellulase enzymatic reaction is: time of enzymatic reacting is 30min, optimal pH is 5.0, enzyme liquid is diluted to 130 times, 50 ℃ of optimum temperutures; The enzyme activity that records under the optimum condition is 1685IU/g.Acidic cellulase once adds, and is undertaken by aseptic technique during interpolation, avoids living contaminants as far as possible; Then at 30 ℃, the 120r/min shake-flask culture, survey acidity every day, when acidity reaches 2% left and right sides, except that not adding Mierocrystalline cellulose, the blank sample do not produce the bacterium seed liquor, all the other samples insert the plain bacterium seed liquor that produces of 2% activation good fiber, and the microbiological contamination phenomenon on simulation is produced is observed the situation that fermented liquid produces film simultaneously.
In experiment, after the CELLULOLYTIC BACTERIUM of access 2%, 0% sample cultivation 1d just has the little group of cellulose silk to occur, and Mierocrystalline cellulose is rolled into a ball obviously and increased during 3d, increases not obvious up to cultivating the end Mierocrystalline cellulose afterwards; There is very broken fiber fines silk to occur during 0.02% sample 3.5d, also has only the little group of Mierocrystalline cellulose to occur until cultivate end afterwards; The phenomenon the same with 0.02% sample appearred during 0.05% sample 5d.
Blank only inserts that isolating acetic bacteria is purebred to be cultivated, and cellulose-less produces; The addition of acidic cellulase is that the acid production speed of 0.02%, 0.05%, 0.08%, 0.1%, 0.3%, 0.5%, 0.8% sample is consistent with blank sample with the final acid amount of producing in the fermented liquid, and the interpolation that cellulase be described does not have to influence to the acetic fermentation of above sample; The product acid amount of 0% sample after 1.5d just begins to be lower than other samples, this may be along with bacteria cellulose produces growth of bacterium and increasing of cellulose amount, consume little acetic acid [Ma Xia, Wang Ruiming, Jia Shiru, Deng. non-carbohydrate influences the pre-test [J] of rule to the synthetic bacteria cellulose of acetobacter xylinum. and China brewages, 2003, (4): 15-17], the quickening of the bacteria growing of bacteria cellulose generation simultaneously can influence the product acid of acetic bacteria, make in the fermenting process and fermentation ends after, the acidity of 0% sample is lower than blank and enzyme-added sample, desmoenzyme is to cellulosic action effect and add the increase that enzyme makes product cost, add 0.08% acidic cellulase and just can effectively prevent and treat the generation of bacteria cellulose film, do not influence simultaneously the acetic fermentation of fermented liquid, and after adding acidic cellulase, whole acetic fermentation process there is not all mycoderm to produce.
The contriver is according to above-mentioned test-results, proposed the solution of control mycoderm in the liquid sprinkling vinegar zymotechnique, and promptly the disposable interpolation of the beginning in acetic fermentation stage is not less than the acidic cellulase of material total mass 0.08% in this technological process; Undertaken by aseptic technique during interpolation, avoid living contaminants.
It is 0.08% of material total mass that above-mentioned interpolation acidic cellulase is preferably measured.
The acidic cellulase of above-mentioned interpolation is the commercially available prod, the optimum condition of this cellulase enzymatic reaction is: time of enzymatic reacting is 30min, and optimal pH is 5.0, and enzyme liquid is diluted to 130 times, 50 ℃ of optimum temperutures, the enzyme activity that records under this optimum condition are 1685IU/g.
The method of preventing and treating of mycoderm in the liquid sprinkling vinegar fermentative production of the present invention, first from output that how to reduce bacteria cellulose or the angle research bacteria cellulose of eliminating at all, and invented the method that mycoderm produces in the control acetic fermentation process, and only concentrate on aspect researchs such as the output that how to improve bacteria cellulose and fermentation kinetics at present in the research aspect this; Use the inventive method, can fundamentally prevent and treat the generation of bacteria cellulose film, guarantee that the long period of liquid sprinkling vinegar fermentative production is normally moved.The inventive method is applicable to the enterprise that adopts liquid sprinkling vinegar zymotechnique.
Embodiment
Embodiment
The acetic bacteria seed liquid that activation is good inserts 50ml by the inoculum size of quality of material 6%, and to take from flavor water shield garden alcoholic strength be in the rice saccharification liquid of 6.3% (volume fraction), makees 3 parallel samples for every group.Wherein do blank for one group, the acidic cellulase (being 0.04g) of the adding material total mass 0.08% of another group, 30 ℃, the 120r/min shake-flask culture is surveyed the acid amount of producing at different incubation times; When acidity reaches 2% left and right sides, the blank sample carries out purebred acetic fermentation, and another group test sample inserts the plain bacterium seed liquor that produces of 1ml activation good fiber, the microbiological contamination phenomenon on simulation is produced, observe the product film situation in the fermented liquid simultaneously, and according to the right amount alcohol of how much adding of producing acid.The enzyme-added sample of blank sample and adding material total mass 0.08% produces up to the equal cellulose-less film of fermentation ends, and both acid production speeds and the final acid amount close (the final acid amount of producing is respectively 8.14% and 8.49%) of producing, both reducing sugar content illustrate the not influence of interpolation Dichlorodiphenyl Acetate fermentation of cellulase also near (being respectively 2.048% and 2.078%); This scheme can be used in the real attenuation production of liquid sprinkling vinegar.
Claims (3)
1. the method for preventing and treating of mycoderm in the liquid sprinkling vinegar fermentative production is characterized in that this method is the acidic cellulase that is not less than material total mass 0.08% in the disposable interpolation of the acetic fermentation stage of this technological process; Undertaken by aseptic technique during interpolation, avoid living contaminants.
2. according to the method for preventing and treating of mycoderm in the described liquid sprinkling vinegar fermentative production of claim 1, the addition that it is characterized in that described acidic cellulase is 0.08% of a material total mass.
3. the method for preventing and treating of mycoderm in the liquid sprinkling vinegar fermentative production according to claim 1, the acidic cellulase that it is characterized in that described interpolation is the commercially available prod, the suitableeest enzymatic reaction condition of this cellulase is: time of enzymatic reacting is 30min, the pH value is 5.0, enzyme liquid is diluted to 130 times, and the enzyme activity that records under 50 ℃ of the temperature is 1685IU/g.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948734A (en) * | 2010-08-27 | 2011-01-19 | 贵州大学 | Method for preventing mycoderm from generating in process of producing vinegar by rice sacchariferous liquor alcohol fermented liquid |
| CN102212589A (en) * | 2011-04-29 | 2011-10-12 | 钟春燕 | Method for preparing bacterial cellulose |
| CN102337320A (en) * | 2011-09-30 | 2012-02-01 | 贵州大学 | New method for producing bacterial cellulose |
| CN102373143A (en) * | 2010-08-25 | 2012-03-14 | 贵州大学 | Method for preventing generation of mycoderm in process of producing vinegar through pouring fermentation |
| CN106029577A (en) * | 2014-02-27 | 2016-10-12 | 巴斯夫欧洲公司 | Process for making fluorinated lithiated mixed transition metal oxides |
| CN106165744A (en) * | 2016-08-17 | 2016-11-30 | 浙江茗皇天然食品开发股份有限公司 | The equipment of continuous static fermentation black tea bacterium solution and manufacture method |
| CN106165742A (en) * | 2016-08-17 | 2016-11-30 | 浙江茗皇天然食品开发股份有限公司 | The production equipment of ferment local-flavor tea concentrated solution and manufacture method |
| CN110172431A (en) * | 2019-07-12 | 2019-08-27 | 贵州大学 | A method of vinegar miscellaneous bacteria is shone in separation, identification |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1618338A (en) * | 2004-12-09 | 2005-05-25 | 叶明伟 | Method for producing beverage of aloes acetic acid |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102373143A (en) * | 2010-08-25 | 2012-03-14 | 贵州大学 | Method for preventing generation of mycoderm in process of producing vinegar through pouring fermentation |
| CN101948734A (en) * | 2010-08-27 | 2011-01-19 | 贵州大学 | Method for preventing mycoderm from generating in process of producing vinegar by rice sacchariferous liquor alcohol fermented liquid |
| CN101948734B (en) * | 2010-08-27 | 2012-09-19 | 贵州大学 | Method for preventing mycoderm from generating in process of producing vinegar by rice sacchariferous liquor alcohol fermented liquid |
| CN102212589A (en) * | 2011-04-29 | 2011-10-12 | 钟春燕 | Method for preparing bacterial cellulose |
| CN102337320A (en) * | 2011-09-30 | 2012-02-01 | 贵州大学 | New method for producing bacterial cellulose |
| CN102337320B (en) * | 2011-09-30 | 2013-04-17 | 贵州大学 | New method for producing bacterial cellulose |
| CN106029577A (en) * | 2014-02-27 | 2016-10-12 | 巴斯夫欧洲公司 | Process for making fluorinated lithiated mixed transition metal oxides |
| CN106165744A (en) * | 2016-08-17 | 2016-11-30 | 浙江茗皇天然食品开发股份有限公司 | The equipment of continuous static fermentation black tea bacterium solution and manufacture method |
| CN106165742A (en) * | 2016-08-17 | 2016-11-30 | 浙江茗皇天然食品开发股份有限公司 | The production equipment of ferment local-flavor tea concentrated solution and manufacture method |
| CN106165742B (en) * | 2016-08-17 | 2023-05-02 | 浙江茗皇天然食品开发股份有限公司 | Production equipment and manufacturing method of fermented flavor tea concentrated solution |
| CN106165744B (en) * | 2016-08-17 | 2023-05-02 | 浙江茗皇天然食品开发股份有限公司 | Continuous static fermentation equipment for black tea fungus liquid and manufacturing method |
| CN110172431A (en) * | 2019-07-12 | 2019-08-27 | 贵州大学 | A method of vinegar miscellaneous bacteria is shone in separation, identification |
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| CN101153260B (en) | 2012-03-28 |
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Effective date of registration: 20151221 Address after: 410600 Ningxiang economic and Technological Development Zone, Hunan, Changsha Patentee after: Jiajia Food Group Co.,Ltd. Address before: 550003 School of chemical engineering, CAI Jie campus, Guizhou University, Guizhou, Guiyang Patentee before: Guizhou University |
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| CB03 | Change of inventor or designer information |
Inventor after: Zhou Shangting Inventor after: Liu Yongjiao Inventor after: Chen Sheng Inventor after: Li Yaobo Inventor after: Fu Yulin Inventor before: Lu Hongmei Inventor before: Zhang Yongfeng |
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