CN101153260B - 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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- CN101153260B CN101153260B CN2007100779203A CN200710077920A CN101153260B CN 101153260 B CN101153260 B CN 101153260B CN 2007100779203 A CN2007100779203 A CN 2007100779203A CN 200710077920 A CN200710077920 A CN 200710077920A CN 101153260 B CN101153260 B CN 101153260B
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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; when the acidic cellulose is added, no bacteria operation is done to avoid the pollution of the bacteria; 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, drench the method for preventing and treating of watering mycoderm in the vinegar fermentative prodn in particular to liquid state.
Background technology
It is widely used in the world advanced vinegar brewing method that fermentation method is watered in the liquid pouring of vinegar; 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; At present China's vinegar liquid drench water in the fermentative Production ubiquitous serious problems be through drench about 1～March water backflow after; 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 fermenting.It is raw material owing to adopting low-concentration ethanol liquid that the fermentation vinegar is watered in the liquid state pouring of external particularly Germany; 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; 1994,1:36-4 3); And China is raw material with the rice, and the product of production is a rice vinegar.The moity 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 and are carried out orderly arrangement to stopping composition; Make stopping composition than in the past more fully by fermented liquid " spray "; The harsh length of mycoderm just swept away and can not gather (to liquid improve one's methods [J] that waters fermentation tower that drench of traditional vinegar. Chinese seasonings, 2001,4:9-3 3); 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 liquid method of preventing and treating of watering mycoderm in the vinegar fermentative prodn of drenching, fundamentally prevent the generation of bacteria cellulose film, guarantee that the liquid LP that waters the vinegar fermentative prodn that drenches normally moves.
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 staple 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 liquid pouring of certain company and water the stopping composition corn cob in the long film fermentation jar in the vinegar fermentative prodn, 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: in liquid fermentation medium, produce the film slant strains with the transfering loop picking, dilution is coated with flat board as stated above, is upside down in then and cultivates the single bacterium colony that homogeneous on flat board, occurs 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, explain 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:
Through qualitative observation Mierocrystalline cellulose method, cellulase hydrolysis method, scanning electron microscopic observation dry film configuration of surface structure method with in the X-ray diffraction method of testing of film the mycoderm component has been carried out qualitative examination; 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 carries out activation; And insert 50ml 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(tric)fraction), and every group parallel does 3 appearance; Do the blank except that one group; Other group adds commercially available acidic cellulase (Sai Lanbo company buys from Kweiyang) 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; Ph optimum is 5.0, and 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 is surveyed acidity every day, when acidity reaches 2% left and right sides; Except that not adding Mierocrystalline cellulose, blank appearance 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, behind 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 is 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, and this possibly be along with bacteria cellulose produces growth of bacterium and increasing of cellulose amount, consumes little acetic acid [Ma Xia; Wang Ruiming, Jia Shiru, etc. 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 bacteria cellulose quickening that produces bacteria growing simultaneously can influence the product acid of acetic bacteria, make in the fermenting process with 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, adds the generation that 0.08% acidic cellulase just can effectively be prevented and treated bacteria cellulose film, does not influence the acetic fermentation of fermented liquid simultaneously; And after adding acidic cellulase, whole acetic fermentation process does not all have mycoderm to produce.
The contriver is according to above-mentioned test-results, proposed the solution that control mycoderm in the vinegar zymotechnique is watered in liquid pouring, 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 ph optimum is 5.0, and enzyme liquid is diluted to 130 times; 50 ℃ of optimum temperutures, the enzyme activity that under this optimum condition, records are 1685IU/g.
The liquid method of preventing and treating of watering mycoderm in the vinegar fermentative prodn of drenching 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 liquid LP that waters the vinegar fermentative prodn that drenches normally moves.The inventive method is applicable to and adopts the liquid enterprise of watering the vinegar zymotechnique of drenching.
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(tric)fraction), does 3 parallel appearance 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, intermittent gauging produces the acid amount when different culture; When acidity reaches 2% left and right sides; Blank appearance is carried out purebred acetic fermentation, and another group test appearance is inserted 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.Blank appearance produces up to the equal cellulose-less film of fermentation ends with the enzyme-added appearance that adds material total mass 0.08%; 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 are explained the not influence of interpolation Dichlorodiphenyl Acetate fermentation of cellulase also near (being respectively 2.048% and 2.078%); This scheme can be used for liquid the pouring in the real attenuation production of watering vinegar.
Claims (3)
1. liquid state is drenched the method for preventing and treating of watering mycoderm in the vinegar fermentative prodn, it is characterized in that being not less than in the disposable interpolation of the acetic fermentation stage of vinegar fermentation production process the acidic cellulase of material total mass 0.08%; Undertaken by aseptic technique during interpolation, avoid living contaminants;
The enzymatic reaction condition of said acidic cellulase is: the reaction times is 30min, and the pH value is 5.0, and enzyme liquid is diluted to 130 times, and the enzyme activity that records under 50 ℃ of the temperature is 1685IU/g.
2. according to the described liquid method of preventing and treating of watering mycoderm in the vinegar fermentative prodn of drenching of claim 1, the addition that it is characterized in that said acidic cellulase is 0.08% of a material total mass.
3. the liquid method of preventing and treating of watering mycoderm in the vinegar fermentative prodn of drenching according to claim 1 is characterized in that described acidic cellulase is the commercially available prod.
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Families Citing this family (8)
| 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 |
| 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 |
| CN102212589B (en) * | 2011-04-29 | 2013-05-22 | 钟春燕 | Method for preparing bacterial cellulose |
| CN102337320B (en) * | 2011-09-30 | 2013-04-17 | 贵州大学 | New method for producing bacterial cellulose |
| EP3110762B1 (en) * | 2014-02-27 | 2018-08-08 | Basf Se | Process for making fluorinated lithiated mixed transition metal oxides |
| 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 |
| 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 |
| CN110172431A (en) * | 2019-07-12 | 2019-08-27 | 贵州大学 | A method of vinegar miscellaneous bacteria is shone in separation, identification |
Citations (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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Patent Citations (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 |
Non-Patent Citations (2)
| Title |
|---|
| 张春枝等.塔醋生产中"海月"的影响及防治.《大连轻工业学院学报》.1997,第16卷(第04期),36-39. |
| 谢恺熙等.对传统食醋液态淋浇发酵塔的改进方法.《中国调味品》.2001,(第04期),32-33、9. * |
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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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