CN101434911B - Wine brewing yeast strain and method for producing ethanol by efficient sugarcane juice fermentation - Google Patents
Wine brewing yeast strain and method for producing ethanol by efficient sugarcane juice fermentation Download PDFInfo
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Abstract
The invention relates to an excellent Saccharomyces cerevisiae gxas02 and a method for carrying out scale production of ethanol by utilizing the Saccharomyces cerevisiae gxas02 as a production strain and adopting sugar cane juice as a raw material. The strain is vacuum freeze-dried and collected in China Center for Type Culture Collection with the collection No. of CCTCC NO: M208110, the collection date is July 19, 2008, the strain is the Saccharomyces cerevisiae by identification, the strain is applicable to fermenting the ethanol by using the sugar cane juice, with fast speed, low residual sugar content, high temperature resistance and alcohol resistance. A production technology adopts a single concentration continuous flow processing technology, the sugar cane juice which is pressed but not concentrated or diluted is directly fermented under the conditions without using yeast reuse and fixed yeast technology, the continuous operation is carried out on a production line with daily production capacity of 10 tons of the ethanol, the effect is good, the fermentation time is 12-16h, alcohol degree reaches 8.6-9.4(V/V) and the residual sugar is 0.2-0.3g/100ml. The invention is characterized by convenient implementation, high equipment utilization rate, high ethanol concentration of fermented mash, high sugar utilization rate which is close to the theoretical value, and the like, thereby reducing the costs for producing the ethanol.
Description
Technical field
The invention belongs to yeast saccharomyces cerevisiae biological fermentation field, specifically is the ethanol fermentation industrial circle, particularly relates to the method for a kind of good Wine brewing yeast strain and producing ethanol by efficient sugarcane juice fermentation thereof.
Background technology
Produce alcohol fuel with saccharine material,, and ignored by people always owing to high raw materials cost.Constantly riseing of crude oil price, and the drop of sugared valency, and the immense success of Brazilian sugarcane alcohol fuel show the good market potential of sugarcane ethanol to common people.South China, located in subtropical zone, temperature height, sunny, rainy and autumn drought, it is historical and formed scale to have a long cultivation of sugar cane, has not only possessed the natural condition of development sugarcane fuel ethanol industrial, has also formed good raw material industry basis simultaneously.Regrettably, at present, do not see the industry application report of mass-producing sugar cane juice fermentative production of ethanol as yet in China.Although sugarcane ethanol also is not the principal mode of present alcohol production,, be necessary it is studied as a redundancy technique.
The more domestic pertinent literature report that research sugarcane alcohol production technology are arranged, as:
Document 1: [author unit] South China Science ﹠ Engineering University bio-science such as fermentation technology optimization [author] Pan Zhuo Zheng Sui Pinglin shadow of energy sugarcane production alcohol fuel and engineering college [periodical name] meet new millennium---the 4th fermentation engineering scientific seminar collection of thesis of industrial biotechnology, the 2006[digest] be raw material with energy sugarcane, " new bridge " yeast is a fermented bacterium, in fresh sugarcane juice, carry out direct fermentation, nitrogenous source interpolation and fermentation condition are studied.Experimental result shows, the interpolation of nitrogenous source can improve zymic growth and fermentation situation, fermentation period is 66h, residual sugar is 4.04g/l, and alcoholic strength is 11.5%, and the suitableeest inoculum size is 10%, the suitableeest fermentation initial pH value is 6.5, the suitableeest leavening temperature is 35 ℃, sends out liquor-saturated can significantly shorten fermentation period under the concussion state, and fermentation period is 32h.
Document 2: seed selection [author] model unit virtuous chapter in river that is fit to sugar cane juice fermentation high yield distillery yeast is old by the industrial microorganism the Ministry of Education of strong grade [author unit] Fujian Normal University Engineering Research Center [periodical name] biotechnology, 2008, (1) [digest] purpose: the bacterial strain that selects the high yield yeast saccharomyces cerevisiae that is fit to fermentation sugar cane juice production alcohol fuel.Method: as starting strain, the protoplastis that obtains behind the enzymolysis broken wall is carried out ultraviolet mutagenesis with yeast strain YS5, carry out seed selection by primary dcreening operation and multiple sieve.Result: obtain the yeast saccharomyces cerevisiae mutant strain YS5-1 that a plant height produces alcohol, the ethanol content of this mutant strain fermentation sugar cane juice can reach 12.6% (V/V), 11.6% (V/V) than starting strain improved 8.6%, and its sugared transformation efficiency is higher than 87.0% of starting strain up to 94.5%.Conclusion: the ethanol production of cultivating its mutant strain of back by 5 continuous passages keeps stable, shows that this mutant strain sugar cane juice that can be used to fully ferment produces alcohol fuel.
Document 3: Li Miaomiao is in refined beautiful [mechanism] South China Science ﹠ Engineering University light industry and Foodstuffs Academy for the current situation of sugarcane production alcohol fuel and prospect forecast [author], and Guangzhou, Guangdong 510640[prints name] brewing science and technology .2007 (6) .-111-113.Mention in the literary composition, abroad, the ethanol factory of Brazil adopts ferments and the two kinds of explained hereafter alcohol fuels that continuously ferment indirectly, and yeast is separated the continuation reuse, and ripe wine with dregs alcohol concn can reach 8%～11%, and fermentation time foreshortens to 6～10h, but reuse in a day 3 times.Zelanian H.W.Doelle1 etc. are raw material with the sugar cane juice; as fermentation strain large-scale production industrial spirit, 100L and 1000L fermentation test show, after fermentation in 17～20 hours with the motion fermentation sporangium; its sugared transformation efficiency is 91～95%, and ethanol concn is 10% (v/v).Limtong, S. etc. are raw material with the sugar cane juice, and kluyveromyces marxianus is that fermentation strain is produced fuel alcohol, anti-40～45 ℃ of high temperature of this yeast, and 40 ℃ of fermentations, ethanol concn is 6.78% (w/v), spirit yield is 60.4%.
Up to the present, the Alcohol Production in sugar refinery all is raw material with molasses, though sugar cane juice is the same as with molasses to contain the sucrose composition, but sugar cane juice has its singularity, less as the sugar cane juice ash content, and protein, colloid etc. are more, and easily causing influences yeast growth, yeast flocculation, fermenting speed partially slowly and problem such as complex production process is easy in the fermenting process.At the singularity of sugar cane juice direct fermentation, select the barms of suitable sugarcane juice fermentation, improve and optimize its zymotechnique, need carry out further investigation in the hope of improving yield aspects.Domestic and international research personnel to the sugarcane alcoholic acid production done a large amount of valuable research work, wherein, Brazil is particularly outstanding, has obtained significant effect, it adopts the fermentation of natural quality yeast, and the combining yeast reuse technology.But at the present situation of China present ethanol factory, carry out the yeast reuse technology, not only equipment claimed is improved, and higher relatively to staff's requirement, how effectively to utilize existing installation to carry out the production of sugarcane alcoholic acid, and the present invention has carried out useful exploration.
Summary of the invention
The objective of the invention is at slow partially, the deficiencies such as residual sugar is high, complex production process of sugar cane juice direct fermentation production ethanol bacterial strain fermenting speed, a kind of good Wine brewing yeast strain of suitable sugarcane juice fermentation is provided, and provide the single concentration Continuous Flow of this bacterial strain in the sugarcane juice producing and ethanol to add fermentation technology, to fill up the blank of China in this production technical field.
Wine brewing yeast strain provided by the present invention (Saccharomyces cerevisiae), be that the contriver utilizes biological means, the strain that isolation and selection obtains from the soil of grain distillery's sewage draining exit is high temperature resistant, ethanol-tolerant, be fit to the bacterial strain of sugarcane fermentation, be yeast strain gxas02, be deposited in the Chinese typical culture collection center that is positioned at Chinese Wuhan City Wuhan University on July 19th, 2008, deposit number is CCTCC NO:M208110.
The isolation and selection process of CCTCC NO:M208110 is as follows:
Gather the earth of depleted mash, waste water, mud and periphery from grain distillery's sewage draining exit and make sample separation, insert respectively and contain in the 250ml triangular flask of 50mlYPD (glucose 20g/L, yeast extract paste 10g/L, peptone 20g/L, natural pH value) liquid, the sample separation that inserts is about 1g, 30 ℃, 160rpm shakes a bottle enrichment culture 24h.Get enrichment culture liquid and carry out the dilution of different concns, be coated with the YPD flat board, cultivated 2 days for 30 ℃, the bacterial strain dilution is coated in the TTC lower floor flat board, cultivate 24h for 30 ℃, grow visible bacterium colony after, selecting colony number is that 100～300 flat board is poured about 12mL TTC upper strata substratum into and covered original bacterium colony, at 30 ℃ of following lucifuge insulation 2～3h, judge that by the colour generation of bacterium colony yeast produces the size of alcohol ability afterwards.By the colour generation characteristic of TTC developer to the yeast metabolism product, the strong yeast of alcohol ability is produced in screening.The contriver according to the method described above, successively from coming from 2 grain distillerys, surplus totally 4 batches 30 in the duplicate samples, select one and belong to of the present invention, have and utilize sugar cane juice to produce the isolate of the good fermentation character of alcohol, gxas 02, i.e. CCTCC NO:M208110 are numbered in the laboratory.
The classification of CCTCC NO:M208110 is identified:
The cell morphological characteristic of CCTCC NO:M208110 is circle or oval, and in the childhood bacterium colony, cell is 4～14 * 3～7 microns, and long and wide ratio is 1: 1～1: 2, in wort, precipitate, and the thecaspore circle, smoothly.
Picking contains the single bacterium colony on the CCTCC NO:M208110 flat board, is inoculated in the finger-type bottle that contains the 3mlYPD substratum 30 ℃, 160rpm, incubated overnight then, utilizes yeast total DNA extraction test kit (day root biochemical technology company limited) to extract the genomic dna of CCTCC NO:M208110.
Wherein adopt:
Forward primer NL-1:5 '-GCATATCAATAAGCGGAGGAAAAG-3 ',
Reverse primer NL-4:5 '-GGTCCGTGTTTCAAGACGG-3 ',
Total DNA with CCTCC NO:M208110 is a template, carry out polymerase chain reaction (PCR), the amplification size is the 26S rDNA D1/D2 zone nucleotide sequence of 575bp, amplified production is connected on the pMD18-T carrier (the precious biotechnology in Dalian company limited), carries out homologous sequence search (Nucleitide-Nucleitide BLAST) with this sequence in Genbank nucleic acid sequence data storehouse.Search Results shows: the 26S rDNA D1/D2 zone nucleotide sequence of CCTCC NO:M208110 all reaches 100% with same regional nucleic acid sequence homologies such as Genbank existing Saccharomyces cerevisiae isolate TJY-29, Saccharomycescerevisiae isolate NY3-4 Saccharomyces cerevisiae isolate NY3-2 and Saccharomycescerevisiae isolate NY3-1.
Because the 26S rDNA D1/D2 zone nucleotide sequence that CCTCC NO:M208110 is simple and other bacterial strains is as broad as long, in order better to distinguish this bacterial strain, the order-checking bacterial strain S288C of this bacterial strain and yeast saccharomyces cerevisiae has been carried out the biochip of CGH (comparative genomic hybridization), and carried out the fluorescence exchange test, search out 271 and the gene of notable difference is arranged with S288C, 23 gene copy number showed increased, mainly concentrate on the 17th karyomit(e), 248 gene copy numbers obviously reduce, and are distributed on remaining 16 karyomit(e).
CCTCC NO:M208110 is carried out temperature and the test of alcohol tolerance level, find this bacterial strain for can tolerate high temperature, high sugar and high alcohol bacterial strain, its temperature tolerance level is 37 ℃, and anti-pol is the glucose of 60% mass ratio, and alcohol tolerance level volume percent is 20%.
According to can determining of above-mentioned bacterium colony outward appearance, cellular form, Physiology and biochemistry and molecular biological characteristics, the classification position of CCTCCNO:M208110 belongs to yeast saccharomyces cerevisiae (Saccharomyces cerevisiae).
The present invention also provides and has utilized CCTCC NO:M208110 direct fermentation sugarcane juice production alcoholic acid list concentration stream to add fermentation process.Concrete processing method is as follows:
CCTCC NO:M208110 is pressed microbial fermentation conventional liq seed culture method, with the YPD substratum, be glucose 20g/L, yeast extract paste 10g/L, peptone 20g/L, nature pH value is cultivated, when containing yeast count, every milliliter of nutrient solution reaches 1.5 hundred million when above, inoculum size by 1～10% is transferred in the sugar cane juice of handling that (having added the vitriol oil, to adjust pH be 3.0～4.0 and to add mass ratio be 0.1% urea, mass ratio is 0.02% phosphoric acid), progressively amplify in this ratio, until can carry out fermentor cultivation, when the fermentation cylinder for fermentation nutrient solution satisfies stream and adds condition, be that every milliliter of nutrient solution contains yeast count and reaches more than 1.5 hundred million, the beginning Continuous Flow adds the sugar cane juice of handling and ferments, technology is controlled the conventional operation parameter of fermentor cultivation liquid yeast number and each fermentor tank routinely, until the karusen maturation, just routinely distil process ethanol production then.
Adopt single concentration stream of the present invention to add fermentation process, in the fermenting process, each jar temperature is controlled at 26～37 ℃, and the used time of fermenting is 12～16 hours; When its fermentative production of ethanol whole process was finished, the ethanol concentration of volume percent was 8.6～9.4% in the fermented liquid, and residual sugar is 0.2～0.3g/100ml.This technology can carry out also can carrying out in a plurality of fermentor tanks in a fermentor tank.
Compared with prior art, outstanding substantive distinguishing features and the obvious improvement that have of the present invention is:
1, isolation and selection has gone out to be suitable for the good fermentation strain of sugarcane juice direct fermentation
The inventor compares by a large amount of screenings, and isolation and selection has gone out to have the Wine brewing yeast strain CCTCCNO:M208110 of multiple good characteristic.The temperature range that this bacterial strain is fit to fermentation is wider, and the ability of anti-high sugar is better, can satisfy the requirement that single concentration stream adds technology, simultaneously, also has characteristics such as fast, the sugared utilization ratio height of fermenting speed, and suitable sugar cane juice is the ethanol fermentation of raw material.
2, production technique has obtained optimization
By seed selection and utilize the excellent fermentation bacterial strain, make the higher single concentration stream of efficient add technology and be achieved.This technology is compared existing pair of concentration stream and is added technology, technology has obtained simplifying greatly, not only technology is easier to control, also saved the input of fermentor tank simultaneously, increase the service efficiency of equipment, reduced production cost, improved production efficiency, and can obtain the karusen of higher ethanol concn, spirit yield is obviously improved.And this technology does not adopt the existing yeast reuse technology of Brazil, and technical process and working method are simplified more.
Description of drawings
Fig. 1 is CCTCC NO:M208110 and Saccharomyces Cerevisiae in S 288C karyomit(e) disparity map relatively.
Fig. 2 is that single concentration stream adds the alcohol prepared by fermenting process flow sheet.
The explanation of preservation information
A kind of Wine brewing yeast strain Saccharomyces cerevisiae gxas02, lyophil preservation (is called for short CCTCC) in China typical culture collection center, the depositary institution address is a Wuhan City Wuhan University, deposit number is CCTCCNO:M208110, and preservation date is on July 19th, 2008.
Embodiment
Below in conjunction with accompanying drawing 2, the present invention is further illustrated by specific embodiment, but do not limit the present invention.
Embodiment one: 20L three fermentor tank sugar cane juice fermenting alcohol lab scales
1, liquid nutrient medium barms cultivation in a small amount
28 ℃ of culture temperature, incubated overnight will be inserted in the liquid YPD substratum at the single bacterium colony of the CCTCC NO:M208110 of the dull and stereotyped preservation of YPD.
2, liquid nutrient medium enlarges the barms cultivation
With the YPD nutrient solution in the step 1 is seed liquor, inoculum size by 1% inserts the sugar cane juice of handling, and (adding vitriol oil adjustment pH is 3.5～4.0, the interpolation mass ratio is 0.1% urea, and mass ratio is 0.02% phosphoric acid), prepare 500ml sugar cane juice nutrient solution.
3,20L three fermentor tank sugar cane juices fermentation
With the sugar cane juice nutrient solution in the step 2 is seed liquor, access contains in No. 1 fermentor tank (volume is 20L) that 10L handled sugar cane juice, ferment, controlling air flow in No. 1 fermentor tank is 100L/h, 30 ℃ of leavening temperatures when the thalline number reaches 1.5 hundred million, begin slow stream and add the sugar cane juice of handling, the control flow velocity, keep the thalline number, when fermentating liquid volume reaches 16L in No. 1 tank body, the interface of No. 1 jar and No. 2 jars is opened, fermented liquid begins to enter jar No. 2, when fermentating liquid volume reaches 16L in No. 2 tank bodies, the interface of No. 2 jars and No. 3 jars to be opened, fermented liquid begins to enter jar No. 3, No. 2,32 ℃ of No. 3 jar leavening temperatures, the thalline number is more than 200,000,000, and total fermentation time is 12h, the karusen maturation, the wine degree reaches 8.6 (V/V), and residual sugar reaches 0.3g/100ml.
Embodiment two: 10 tons of ethanol pilot scales of sugar cane juice fermentation daily output
1, cultivation of liquid nutrient medium a small amount of barms and liquid nutrient medium enlarge the barms cultivation with embodiment one.
2, with the sugar cane juice be fermenting raw materials, produce 10 tons of ethanol pilot scales daily.
With reference to accompanying drawing 2, behind 50L, the fresh seeds liquid of gained inserts in the fermentor tank of handling that contains 10 tons of sugar cane juices (volume is 120 cubic metres) and ferments with CCTCC NO:M208110 bacterial strain enlarged culturing.The sugar cane juice treatment process is: adding vitriol oil adjustment pH in sugarcane juice is 3.0～4.0, and the interpolation mass ratio is 0.1% urea, and mass ratio is 0.02% phosphoric acid.In No. 1 jar, control air flow, 26 ℃ of leavening temperatures, when the thalline number reaches 1.5 hundred million, beginning slowly pumps into the same sugar cane juice of handling to No. 1 jar, the control flow velocity, it is constant to keep the thalline number, when fermented liquid liquid level in No. 1 tank body reaches communicating pipe with No. 2 jars, the interface of No. 1 jar and No. 2 jars is opened, and fermented liquid is because the driving of potential energy begins to enter jar No. 2, control thalline number, when fermented liquid liquid level in No. 2 tank bodies reaches communicating pipe with No. 3 jars, the interface of No. 2 jars and No. 3 jars to be opened, the fermented liquid fermented liquid is because the driving of potential energy, begin to enter jar No. 3, after treating the karusen maturation, just send into distillation tower and distill by prior art and produce alcohol, carbonic acid gas is by unified discharge of pipeline of fermentor tank top.28 ℃ of all the other each jar leavening temperatures of whole process, thalline number are more than 1.5 hundred million, and total fermentation time is 12～16h, and karusen maturation, ethanol concn reach 8.6% (V/V), and residual sugar reaches 0.3g/100ml.
Embodiment three: 10 tons of ethanol pilot scales of sugar cane juice fermentation daily output
1, cultivation of liquid nutrient medium a small amount of barms and liquid nutrient medium enlarge the barms cultivation with embodiment one.
2, with the sugar cane juice be fermenting raw materials, produce 10 tons of ethanol pilot scales daily.
With reference to accompanying drawing 2, behind 50L, the fresh seeds liquid of gained inserts in the fermentor tank of handling that contains 10 tons of sugar cane juices (volume is 120 cubic metres) and ferments with CCTCC NO:M208110 bacterial strain enlarged culturing.The sugar cane juice treatment process is: adding vitriol oil adjustment pH in sugarcane juice is 3.0～4.0, and the interpolation mass ratio is 0.1% urea, and mass ratio is 0.02% phosphoric acid.In No. 1 jar, control air flow, 29 ℃ of leavening temperatures, when the thalline number reaches 200,000,000, beginning slowly pumps into the same sugar cane juice of handling to No. 1 jar, the control flow velocity, it is constant to keep the thalline number, when fermented liquid liquid level in No. 1 tank body reaches communicating pipe with No. 2 jars, the interface of No. 1 jar and No. 2 jars is opened, and fermented liquid is because the driving of potential energy begins to enter jar No. 2, control thalline number, when fermented liquid liquid level in No. 2 tank bodies reaches communicating pipe with No. 3 jars, the interface of No. 2 jars and No. 3 jars to be opened, the fermented liquid fermented liquid is because the driving of potential energy, begin to enter jar No. 3, after treating the karusen maturation, just send into distillation tower and distill by prior art and produce alcohol, carbonic acid gas is by unified discharge of pipeline of fermentor tank top.31 ℃ of all the other each jar leavening temperatures of whole process, thalline number are more than 200,000,000, and total fermentation time is 12～16h, and karusen maturation, ethanol concn reach 9.4% (V/V), and residual sugar reaches 0.2g/100ml.
Embodiment four: 10 tons of ethanol pilot scales of sugar cane juice fermentation daily output
1, cultivation of liquid nutrient medium a small amount of barms and liquid nutrient medium enlarge the barms cultivation with embodiment one.
2, with the sugar cane juice be fermenting raw materials, produce 10 tons of ethanol pilot scales daily.
With reference to accompanying drawing 2, behind 50L, the fresh seeds liquid of gained inserts in the fermentor tank of handling that contains 10 tons of sugar cane juices (volume is 120 cubic metres) and ferments with CCTCC NO:M208110 bacterial strain enlarged culturing.The sugar cane juice treatment process is: adding vitriol oil adjustment pH in sugarcane juice is 3.0～4.0, and the interpolation mass ratio is 0.1% urea, and mass ratio is 0.02% phosphoric acid.In No. 1 jar, control air flow, 32 ℃ of leavening temperatures, when the thalline number reaches 200,000,000, beginning slowly pumps into the same sugar cane juice of handling to No. 1 jar, the control flow velocity, it is constant to keep the thalline number, when fermented liquid liquid level in No. 1 tank body reaches communicating pipe with No. 2 jars, the interface of No. 1 jar and No. 2 jars is opened, and fermented liquid is because the driving of potential energy begins to enter jar No. 2, control thalline number, when fermented liquid liquid level in No. 2 tank bodies reaches communicating pipe with No. 3 jars, the interface of No. 2 jars and No. 3 jars to be opened, the fermented liquid fermented liquid is because the driving of potential energy, begin to enter jar No. 3, after treating the karusen maturation, just send into distillation tower and distill by prior art and produce alcohol, carbonic acid gas is by unified discharge of pipeline of fermentor tank top.34 ℃ of whole each jar of process leavening temperatures, thalline number are more than 200,000,000, and total fermentation time is 12～16h, and karusen maturation, ethanol concn reach 9.1% (V/V), and residual sugar reaches 0.25g/100ml.
Embodiment five: 10 tons of ethanol pilot scales of sugar cane juice fermentation daily output
1, cultivation of liquid nutrient medium a small amount of barms and liquid nutrient medium enlarge the barms cultivation with embodiment one.
2, with the sugar cane juice be fermenting raw materials, produce 10 tons of ethanol pilot scales daily.
With reference to accompanying drawing 2, behind 50L, the fresh seeds liquid of gained inserts in the fermentor tank of handling that contains 10 tons of sugar cane juices (volume is 120 cubic metres) and ferments with CCTCC NO:M208110 bacterial strain enlarged culturing.The sugar cane juice treatment process is: adding vitriol oil adjustment pH in sugarcane juice is 3.0～4.0, and the interpolation mass ratio is 0.1% urea, and mass ratio is 0.02% phosphoric acid.In No. 1 jar, control air flow, 35 ℃ of leavening temperatures, when the thalline number reaches 200,000,000, beginning slowly pumps into the same sugar cane juice of handling to No. 1 jar, the control flow velocity, it is constant to keep the thalline number, when fermented liquid liquid level in No. 1 tank body reaches communicating pipe with No. 2 jars, the interface of No. 1 jar and No. 2 jars is opened, and fermented liquid is because the driving of potential energy begins to enter jar No. 2, control thalline number, when fermented liquid liquid level in No. 2 tank bodies reaches communicating pipe with No. 3 jars, the interface of No. 2 jars and No. 3 jars to be opened, the fermented liquid fermented liquid is because the driving of potential energy, begin to enter jar No. 3, after treating the karusen maturation, just send into distillation tower and distill by prior art and produce alcohol, carbonic acid gas is by unified discharge of pipeline of fermentor tank top.37 ℃ of whole each jar of process leavening temperatures, thalline number are more than 200,000,000, and total fermentation time is 12～16h, and karusen maturation, ethanol concn reach 8.6% (V/V), and residual sugar reaches 0.3g/100ml.
Sequence table
＜110〉Guangxi Academy Of Sciences
＜120〉method of a kind of Wine brewing yeast strain and producing ethanol by efficient sugarcane juice fermentation thereof
<160>1
<210>1
<211>575
<212>DNA
＜213〉yeast saccharomyces cerevisiae (Saccharomyces cerevisiae)
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ggataaatcc?ataggaatgt?agcttgcctc?ggtaagtatt?atagcctgtg?500
ggaatactgc?cagctgggac?tgaggactgc?gacgtaagtc?aaggatgctg?550
gcataatggt?tatatgccgc?ccgtc 575
Claims (5)
1. the sugar cane juice that can ferment is produced alcoholic acid and is produced bacterium, classification called after S. cervisiae Saccharomycescerevisiae, bacterial strain number is gxas02, be from the sewage draining exit of grain distillery, to gather to separate to obtain, lyophil preservation is in China typical culture collection center, deposit number is CCTCC NO:M208110, and preservation date is on July 19th, 2008.
2. production bacterium as claimed in claim 1, it is characterized in that: the 26S rDNA D1/D2 zone of described bacterial strain has the nucleotide sequence shown in the sequence 1, all reaches 100% with the existing Saccharomyces cerevisiae the same area nucleic acid sequence homology of Genbank.
3. production bacterium as claimed in claim 1, it is characterized in that: described bacterial strain and Genbank go up the yeast saccharomyces cerevisiae order-checking bacterial strain Saccharomyces cerevisiae S288C that announces and compare on dna level, there are 271 genes to have significant difference, wherein, 23 gene copy number showed increased, mainly concentrate on the 17th (plastosome) karyomit(e), 248 gene copy numbers obviously reduce, and are distributed on remaining 16 karyomit(e).
4. production bacterium as claimed in claim 1 is characterized in that: described bacterial strain is for can tolerate high temperature and high alcohol bacterial strain, and its temperature tolerance level is 37 ℃, and alcohol tolerance level volume percent is 20%, and glucose tolerance level mass ratio is 60%.
5. utilize the described production of claim 1 bacterium fermentation sugar cane juice to produce the alcoholic acid method, it is characterized in that production process is as follows: CCTCC NO:M208110 is pressed microbial fermentation conventional liq seed culture method, promptly with the YPD substratum, be glucose 20g/L, yeast extract paste 10g/L, peptone 20g/L, nature pH value is cultivated, when containing yeast count, every milliliter of nutrient solution reaches 1.5 hundred million when above, 1～10% inoculum size is transferred to and has added the vitriol oil to adjust pH be 3.0～4.0 and add the urea of mass ratio 0.1% by volume, in the sugar cane juice of the phosphoric acid of mass ratio 0.02%, and 1～10% inoculative proportion progressively amplifies by volume, until can carry out fermentor cultivation, when the fermentation cylinder for fermentation nutrient solution satisfies stream and adds condition, be that every milliliter of nutrient solution contains yeast count and reaches more than 1.5 hundred million, the beginning Continuous Flow adds the sugar cane juice of handling and ferments, technology is controlled the conventional operation parameter of fermentor cultivation liquid yeast number and each fermentor tank routinely, until the karusen maturation can be distilled ethanol production according to routine techniques then; In the fermenting process, leavening temperature is controlled at 26～37 ℃, and fermentation time is 12～16 hours.
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| CN101962620A (en) * | 2010-10-13 | 2011-02-02 | 广西大学 | Saccharomyces cerevisiae strain and application thereof |
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| CN101580790B (en) * | 2009-05-26 | 2012-07-18 | 福建师范大学 | Method for brewing cane white wine with low carbinol content |
| CN101633937B (en) * | 2009-08-20 | 2012-05-23 | 华南理工大学 | Alcohol preparation method by fermenting cane juice with foamed ceramic immobilized yeast |
| CN102634464B (en) * | 2012-03-08 | 2013-04-24 | 江苏今世缘酒业股份有限公司 | High-temperature-resistant saccharomyces cerevisiae and separating and culturing method thereof |
| CN102851320A (en) * | 2012-08-24 | 2013-01-02 | 太仓市周氏化学品有限公司 | Fermentation method for producing ethanol by immobilized yeast |
| CA2938150A1 (en) * | 2014-03-21 | 2015-09-24 | Novozymes A/S | Processes for producing ethanol and yeast |
| CN105732132A (en) * | 2016-01-20 | 2016-07-06 | 杭州富阳高博信息技术服务有限公司 | Method for effectively decomposing agricultural waste by using composite microbial preparation |
| CN105524870A (en) * | 2016-01-20 | 2016-04-27 | 杭州富阳高博信息技术服务有限公司 | Environmentally-friendly preparation product capable of promoting straw degradation |
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| CN101962620A (en) * | 2010-10-13 | 2011-02-02 | 广西大学 | Saccharomyces cerevisiae strain and application thereof |
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