CN102268383B - Yeast used for brewing grape vinegar and preparation method for yeast - Google Patents
Yeast used for brewing grape vinegar and preparation method for yeast Download PDFInfo
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- CN102268383B CN102268383B CN 201110258971 CN201110258971A CN102268383B CN 102268383 B CN102268383 B CN 102268383B CN 201110258971 CN201110258971 CN 201110258971 CN 201110258971 A CN201110258971 A CN 201110258971A CN 102268383 B CN102268383 B CN 102268383B
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Abstract
The invention discloses yeast used for brewing grape vinegar, which is characterized in that: the yeast is Pichia with the collection number of CGMCCNo.3076, and is classified and named as Pichia pastoris, and the strain has been collected in China General Microbiological Culture Collection Center (CGMCC) on May 22, 2009. The Pichia aceticbacteria can ferment a grape wine into grape vinegar, a defect that raw materials are influenced and limited by seasons in the conventional process of producing grape vinegar can be overcome, a fermentation period can be shortened, the production cost is reduced, and high-quality grape vinegar with high acetic acid content and low alcohol content is obtained. The grape vinegar keeps the nutrient components of grape, and has the effect of vinegar, and other additives are not needed to be added in the whole brewing process.
Description
Technical field
The present invention relates to microbial technology field, specifically a kind of yeast for brewing grape vinegar and preparation method thereof.
Background technology
Grape is the fruit of vitaceae, is defoliation liana, is one of ancient planting plants of Most.Grape originates in Europe, West Asia and north African one band, and all there is production existing various places, China north and south, mainly originate in the ground such as Xinjiang, Gansu, Shanxi, Hebei, Shandong, Jiangsu and Zhejiang Provinces and Yunnan.Volume tartaric acid in the grape helps digestion, and the energy invigorating the spleen and regulating the stomach is brewed grape into quantity, the breeding of help probiotics that grape vinegar can reduce undesired bacteria in the enteron aisle, eliminates skin splash.In addition, grape vinegar is physiological alkalinity food, and polysaccharide, the potassium ion in the grape vinegar can reduce the acidity in the body, thus alleviating fatigue, physical strength reinforcing.
Reported at present the method for many manufacturing grape vinegars both at home and abroad, the domestic reports that had much about the grape vinegar preparation:
Be spontaneous fermentation such as its method of application number 200610048850.4 " preparation method of grape vinegar ", homogeneity of product is difficult to guarantee, acetic acid content is low, can only seasonally produce etc.
Be to add the modulation such as honey, white sugar, Sorbic Acid with Sucus Vitis viniferae such as its method of application number 20071005808.5 " preparation method of grape vinegar ", do not brewage.
Be must, acetin such as its method of application number 97104377.9 " a kind of preparation method of grape vinegar ", white sugar etc. mix system, are not fermentation and brewage.
Such as application number 99122634.8 " a kind of hill grape vinegar and manufacture method thereof ", this vinegar is that pure grain brewing vinegar and Vitis Amurensis out is that raw material is made grape vinegar.
Such as application number 200910073703.6 " the grape vinegar method is produced in grape and grain mixed fermentation ", what its method was produced is not pure grape vinegar.
The disclosed method of these patent documentations all has a common ground, be exactly its raw material all take grape fruit or Sucus Vitis viniferae as raw material, mix vinegar, Sorbic Acid, the manufacturing grape vinegar that perhaps forms after the grain fermentation.Yet there are no report is raw material with grape wine directly, and in addition the pichia spp acetic bacteria ferments, and produces high-quality grapes vinegar.
Summary of the invention
The purpose of this invention is to provide a kind of yeast for brewing grape vinegar and preparation method thereof.
Yeast of the present invention is Pichia yeast, and its preservation registration number is: CGMCC No.3076, and Classification And Nomenclature is: pichia pastoris (
Pichia pastoris), this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on May 22nd, 2009.
Saccharomycetic preparation method of the present invention is as follows:
1) acquisition of bacterial classification:
Grape wine is exposed in the air time one length will souring, the acetic bacteria of Here it is grape wine and occurring in nature reacts, perhaps in the wine making process, after sugar part all is fermented into alcohol in the grape fresh fruit, in time do not stop fermentation, slowly grape wine also just turns sour, and to become be the sour wine of the non-vinegar of vinegar, and this also is the acetic acid reaction that airborne acetic bacteria and grape wine occur, the pichia spp acetic bacteria that utilizes this phenomenon to obtain, obtain with the separation and Culture program as follows:
The new fresh wine (do not stop fermentation and namely do not add sulfurous gas) that fresh fruit fermentation is obtained, put into large wide-mouth tinted shade around ring, the degree of depth is 1/5 of wide-mouth, the above builds with gauze, be placed on the ventilation lucifugal place, treat that wine upwards assembles one deck white film, this film is exactly usually to be the vinegar film, from wherein optimize purity high, produce the strain Acetobacter xylinum more than the acid;
2) above-mentioned bacterial classification is moved on to be diluted to 10 in the sterilized water-
4～10-
5Doubly, preparation plate isolation base: glucose 10%, peptone 2%, yeast extract paste 1%, agar 1.5% adds the grape wine to 100% that will brewage; Substratum adds first 4% alcohol under aseptic condition, will be diluted to 10-
4～10-
5The acetic bacteria 1mL of times concentration is divided into 0.1mL point and drops on the substratum, delivers to 30 ℃ of thermostat containers and cultivates, cultivate to occur the acetic bacteria bacterium colony after 5 days, chooses single colony diameter maximum, taking-up redilution to 10-
4～10-
5Doubly, repeat again to cultivate three times;
3) with above-mentioned 2) step grow faster bacterial classification take out with sterilized water be diluted to 10-
4～10-
5Doubly, carry out blotting and cultivate, the blotting substratum forms: glucose 10%, peptone 2%, yeast extract paste 1%, Caco
32%, agar 1.5%, the grape wine to 100% that adding will be brewageed; To be diluted to 10-
4～10-
5The bacterial classification of times concentration drops in by every 0.1mL point that (multipotency drips 5 points on each substratum on the blotting substratum for preparing, leave enough distances between every), substratum was put into 30 ℃ of thermostat containers cultivations after 3～5 days, acid producing ability is strong, and the acetic acid bacteria strain of robust growth is selected and is carried out secondary blotting and cultivate, after cultivating to 3～4 times blottings, the acetic acid bacteria strain that output is stable, namely pichia pastoris (
Pichia pastoris).
This culture propagation preserved or through I level II level spawn culture expand numerous after for the production of.
The morphological specificity of kind of bacterium: observe bacterial classification with ordinary optical microscope, cellular form is avette, and this bacterium vegetative propagation mode is the cell budding, pseudohypha, sexual propagation mode are arranged is to produce ascus, and ascus is half garden shape, smooth surface.Bacterium colony is oyster white, and circle is dry matt, is easily provoked on substratum.
Molecular Identification: extract yeast sclerotium DNA, carry out the pcr amplification of 18SrDNA with the yeast universal primer, the PCR product checks order after being connected to the pMD18-T carrier, sequencing result is carried out BLAST at NCB1 analyze, the result shows that having 99% with pichia sp.NRRL Y-27259 bacterium has homology.
Characteristic:
28 ℃～33 ℃ of the sour temperature of the suitableeest life;
Alcohol average conversion 93%～95%;
The highest acidproof amount 9～11(is with acetometer).
Utilize pichia spp acetic bacteria of the present invention wine fermentation can be become grape vinegar produce a kind of grape vinegar, can overcome the shortcoming that existing grape vinegar production process Raw is subjected to the seasonal effect restriction, and can shorten fermentation period, Decrease production cost, obtain top-quality vinegar acidity, the high-quality grapes vinegar of low ethanol content.The nutrition that has namely kept grape to have has again the effect of vinegar, and whole brewing process need not to add other additive.
Pichia yeast of the present invention is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, address on May 22nd, 2009: Datun Road, Chaoyang District, Beijing City China institute of microbiology.
Embodiment
Saccharomycetic preparation method of the present invention is as follows:
1) acquisition of bacterial classification:
The new fresh wine (do not stop fermentation and namely do not add sulfurous gas) that fresh fruit fermentation is obtained, put into large wide-mouth tinted shade around ring, the degree of depth is 1/5 of wide-mouth, the above builds with gauze, be placed on the ventilation lucifugal place, treat that wine upwards assembles one deck white film, this film is exactly usually to be the vinegar film, from wherein optimize purity high, produce the strain Acetobacter xylinum more than the acid;
2) above-mentioned bacterial classification is moved on to be diluted to 10 in the sterilized water-
4～10-
5, preparation plate isolation base forms: glucose 10%, and peptone 2%, yeast extract paste 1%, agar 1.5% adds the grape wine to 100% that will brewage; Substratum adds first 4% alcohol under aseptic condition, will be diluted to 10-
4Concentration acetic bacteria 1mL be divided into 0.1mL point and drop on the substratum, deliver to 30 ℃ of thermostat containers and cultivate, cultivate and occur the acetic bacteria bacterium colony after 5 days, choose single colony diameter maximum, taking-up is diluted to 10-
4, repeat again to cultivate three times;
3) with above-mentioned 2) bacterial classification of growing faster take out with sterilized water be diluted to 10-
4, carrying out blotting and cultivate, the blotting substratum forms: glucose 10%, peptone 2%, yeast extract paste 1%, Caco
32%, agar 1.5%, the grape wine to 100% that adding will be brewageed; To be diluted to 10-
4The bacterial classification of concentration drops in by every 0.1mL point that (multipotency drips 5 points on each substratum on the blotting substratum for preparing, leave enough distances between every), substratum was put into 30 ℃ of thermostat containers cultivations after 3～5 days, acid producing ability is strong, and the acetic bacteria of robust growth, this bacterial classification is selected the blotting that carries out second period cultivate, to 3～4 all after dates, the acetic acid bacteria strain that output is stable, its preservation registration number is: CGMCC No.3076, Classification And Nomenclature is: pichia pastoris (
Pichia pastoris).
This culture propagation preserved or through I level II level spawn culture expand numerous after for the production of.
Claims (1)
1. a yeast that is used for brewing grape vinegar is characterized in that described yeast is Pichia yeast, and its preservation registration number is: CGMCC No.3076, and Classification And Nomenclature is: the pichia pastoris bacterium (
Pichia pastoris), this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on May 22nd, 2009.
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| CN 201110258971 CN102268383B (en) | 2011-09-05 | 2011-09-05 | Yeast used for brewing grape vinegar and preparation method for yeast |
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| CN102268383B true CN102268383B (en) | 2013-03-27 |
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| CN108034610B (en) * | 2018-01-04 | 2019-09-27 | 四川缪氏现代农业发展有限公司 | Grape vinegar aoxidizes acetic acid bacteria and its application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101130742A (en) * | 2007-08-01 | 2008-02-27 | 陈士民 | Preparation method of grape vinegar |
| CN101481650A (en) * | 2009-01-20 | 2009-07-15 | 山西省食品工业研究所 | Method for producing grape vinegar by mixed fermentation of grape and grain |
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2011
- 2011-09-05 CN CN 201110258971 patent/CN102268383B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101130742A (en) * | 2007-08-01 | 2008-02-27 | 陈士民 | Preparation method of grape vinegar |
| CN101481650A (en) * | 2009-01-20 | 2009-07-15 | 山西省食品工业研究所 | Method for producing grape vinegar by mixed fermentation of grape and grain |
Non-Patent Citations (3)
| Title |
|---|
| Solieri，L et al..Molecular assessment of indigenous yeast population from traditional balsamic vinegar.《Journal of applied microbiology》.2006,第101卷(第1期),63-71. * |
| 杨晓红.葡萄发酵饮料的研制.《陕西师范大学学报(自然科学版)》.2002,第30卷175-177. * |
| 武运 等.葡萄果醋酒精发酵过程工艺的研究.《中国酿造》.2010,(第3期),113-115. * |
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Address after: 675000 Yunnan Yi Autonomous Prefecture in Chuxiong province Yongren County Lian Chi Xiang Yao Cun Charlie. Patentee after: Yunnan and biotechnology Limited by Share Ltd. Address before: 675000 Yunnan Yi Autonomous Prefecture in Chuxiong province Yongren County Lian Chi Xiang Yao Cun Charlie. Patentee before: Yunnan and Li Biotechnology Co.,Ltd. Address after: 675000 Yunnan Yi Autonomous Prefecture in Chuxiong province Yongren County Lian Chi Xiang Yao Cun Charlie. Patentee after: Yunnan and Li Biotechnology Co.,Ltd. Address before: 675000 Yunnan Yi Autonomous Prefecture in Chuxiong province Yongren County Lian Chi Xiang Yao Cun Charlie. Patentee before: YUNNAN YONGREN HELI GRAPE VINEGAR BREWING Co.,Ltd. |