CN103820346B - One Accharomyces cerevisiae and the application in fermentation producing and ethanol thereof - Google Patents
One Accharomyces cerevisiae and the application in fermentation producing and ethanol thereof Download PDFInfo
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Abstract
The invention discloses an Accharomyces cerevisiae and the application in fermentation producing and ethanol thereof, yeast saccharomyces cerevisiae (<i>SaccharomycesLEssT.LTssT.L T/i><i>? cerevisiae</i>) SC-X234, be preserved in China typical culture collection center CCTCC on January 6th, 2014, deposit number: CCTCC? M? 2014005, have employed low energy N
+inject mediation spt15 gene engineering, with SC-100 for the bacterium that sets out, utilize? TTC is dull and stereotyped, alcohol is dull and stereotyped carries out primary dcreening operation, the bacterial strain obtained carries out the multiple sieve that ferments, improve the patience of yeast in alcohol gradually, final acquisition output can reach the production bacterial strain SC-X234 of 15.75%, and this bacterial strain can improve the low problem of fermentation production rate greatly, and sugar alcohol transformation efficiency is higher.In 5L fermentor tank, ethanol production can reach 126g/l, improves 87.5%, have very high economic worth as alcohol fuel than the original bacterium that sets out.
Description
Technical field
The invention belongs to technical field of microbial fermentation, be specifically related to an Accharomyces cerevisiae and the application in fermentation producing and ethanol thereof.
Background technology
Fossil energy is the main body of current energy structure, and be the Nonrenewable resources that a class is very valuable, its reserves are limited, for China, though China is oil production big country, is oil consumption big country and petroleum resources shortage big country also.Within 2008, China produces crude oil 1.9 hundred million tons; Consumption 3.65 hundred million tons, oil, net importation crude oil 1.75 hundred million tons, import volume accounts for 48% of total quantity consumed.And by the end of the year 2006, China's accumulation verified oil workable reserve is 20.43 hundred million tons, calculate like this, existing oil only can maintain the needs of decades, and contradiction between oil supply and demand is very outstanding, and will constantly aggravate.Under this overall situation, carry out the exploitation of biomass energy, replace non-renewable fossil energy just to have important practical significance.The current Chinese government has also been fully recognized that the importance of application this " green energy resource ", in recent years constantly increases the investment of biomass energy research, particularly substitutes grain-production alcohol fuel to research and development vegetable fibre and gives great expectations.
Alcohol fuel (fuelethanol) points in gasoline or diesel oil to add a certain proportion of dehydrated alcohol.One solves the potential a limited number of problem as primary energy source gasoline, diesel oil; Two is the burning levels improving gasoline and diesel oil, is conducive to strengthen environmental protection.Although the fuel value of alcohol is only 2/3 of gasoline combustion value, containing aerobic in alcohol molecule, in combustion processes, the capability of antidetonance is good.When in gasoline, the addition of alcohol is no more than 15%, the rideability of automobile is had no significant effect, but the content of CO, NOx and CH compound in tail gas can reduce by 30% ~ 50%.On technical research, the exploitation that huge manpower and materials carry out alcohol fuel has all been dropped in countries in the world, and especially to substitute grain-production alcohol fuel advantage remarkable for research and development vegetable fibre.For the U.S., the U.S. with the technological development of Mierocrystalline cellulose ethanol comparatively early, USDOE is submitted a plan for 1999, by 2005, the production cost of alcohol fuel is reduced by 36%, 1999, renewable energy source institute of the U.S. and reached quite high level about the crucial fermentation technique of develop enterprise, remains certainly in some problems.Current various countries scientist achieves major progress in a lot of fields: the production efficiency as microbial cellulase significantly improves, and multiple new raw materials pretreatment practical technique is developed; The engineering bacteria that can be total to glucose fermentation and wood sugar successfully builds; The research of straw biological transforming fuel alcohol reactor obtains certain progress; The pilot scale of cellulose raw producing and ethanol has been extended to tens of cubic metres of fermentor tank industrial scales etc.Utilize the agricultural crop straw of enormous amount and forest products tankage (lignocellulose-containing) to produce alcohol fuel, list " 15 ", " 11th Five-Year " development in science and technology of China in the works.Utilizing cellulosic material to produce ethanol is utilize the microorganism of cellulase-producing or cellulase first cellulose hydrolysis to be become fermentable sugar, recycling yeast is fermented into ethanol, therefore the bacterial strain obtaining efficient producing and ethanol is key factor wherein, and various screening high-yield ethanol bacterial strain becomes the focus of research simultaneously.Improve herein by describing a kind of novel mutafacient system the ability that bacterial strain produces alcohol, improve the patience of yeast saccharomyces cerevisiae to ethanol simultaneously.The highest level of current domestic producing and ethanol is 12%, and external highest level can to 15%.
Summary of the invention
Low in order to solve ethanol production in the producing and ethanol that ferments at present, there is no the bacterial strain of high-yield ethanol, and then the problem that fermentation producing and ethanol production cost is high, the invention provides an Accharomyces cerevisiae SC-X234 and the application in fermentation producing and ethanol thereof, improve yeast saccharomyces cerevisiae to the patience of alcohol in fermentation by mutagenesis, ensure simultaneously ethanol production 15.75% level.
In order to achieve the above object, the technical solution used in the present invention is: a kind of yeast saccharomyces cerevisiae (
saccharomycescerevisiae) SC-X234, be preserved in China typical culture collection center CCTCC, deposit number on January 6th, 2014: CCTCCM2014005, address: Wuhan, China Wuhan University.
The screening method of Saccharomyces Cerevisiae in S C-X234 of the present invention: the original starting strain SC-100 of yeast saccharomyces cerevisiae injects after mediation spt15 gene through ion beam of low energy N+, utilize the bacterial strain that high ethanol is dull and stereotyped, TTC plate screening obtains, after resistance to ethanol domestication and shake flask fermentation obtains the highest yeast saccharomyces cerevisiae of producing and ethanol concentration is next round mutagenic strain.Repeat above-mentioned steps, until obtain aimed strain SC-X234, concrete steps are as follows:
A the preparation of () monospore suspension: the bacterium SC-100 that set out by yeast saccharomyces cerevisiae makes spore suspension, concentration is 1 × 10
6individual/mL;
The mutagenesis of (b) Mediated by Low Energy Ion Beam spt15 gene: get 100 μ L spore suspensions and the 50 μ L plasmid (plasmid containing spt15 is permitted beautiful jade teacher by Nanjing University of Technology and given) containing spt15, even spread is on aseptic empty flat board, dry up with sterile wind, under 20Kev energy, 40 × 10
14ions/cm
2carry out Nitrogen ion mutagenesis under dosage, after ion implantation, carry out wash-out in an aseptic environment with 1mLTE solution, the bacterium liquid after wash-out, at 37 DEG C of temperature bath 1h, is then applied on alcohol flat board, carries out pressure screening, be inverted for 37 DEG C and cultivate 2-3d.
C the primary dcreening operation of () TTC flat board: be poured on by TTC upper strata substratum low temperature on the flat board grown in step (b), be inverted at 37 DEG C and cultivate 1-2d, screen the bacterial strain that cell dehydrogenase activity is strong, vigor is stronger, and redness is darker.
The multiple sieve of (d) fermentation: S. cervisiae access inclined-plane step (c) filtered out, cultivate 1-2d at 37 DEG C, then inclined-plane proceeds in seed culture medium, cultivates 8-10h at 37 DEG C.Get in seed liquor access fermention medium, inoculum size 10%(v/v), 250mL shaking flask dress 30mL substratum, leavening temperature 37 DEG C, fermentation 48h, measure the content of alcohol in fermented liquid, filter out the starting strain of the highest S. cervisiae of ethanol content as next round mutagenesis screening, repeat above-mentioned steps until screen aimed strain SC-X234.
In above-mentioned screening method: the alcohol flat board in step (b) is the alcohol adding 5%-10% in YPD substratum, the TTC substratum of step (c) is divided into upper strata substratum and lower floor's substratum, lower floor's substratum is YPD substratum, upper strata nutrient media components is that TTC0.5g glucose 5g agar 15g constant volume is to 1L, pH nature.Step (d) seed culture based component carbon source 5%, nitrogenous source 0.7%, inorganic salt 0.016%, buffer reagent 0.1%, pH=5.5, fermentation medium components composition in step (d): carbon source 36%, nitrogenous source 1.1%, inorganic salt 0.02%, buffer reagent 0.5%, wherein carbon source is one in glucose or W-Gum or mixture; Nitrogenous source: organic nitrogen source is yeast extract paste, inorganic nitrogen-sourced: ammonium sulfate, pH=5.5, buffer reagent is primary ammonium phosphate.
Yeast saccharomyces cerevisiae of the present invention (
saccharomycescerevisiae) morphology of SC-X234 and physiochemical properties:
Colony colour: oyster white
Aerobic mode: amphimicrobian
Bacterium colony size: 2-3mm
Growth temperature: 35-40 DEG C
Optimal pH: 5.0-5.5
Colonial morphology: circular
Methylene blue dyes: viable bacteria is colourless, and dead bacterium is blue.
The application of above-mentioned Saccharomyces Cerevisiae in S C-X234 in fermentative production of ethanol, comprises the steps:
1) slat chain conveyor: Saccharomyces Cerevisiae in S C-X234 is connected on dull and stereotyped minimum medium and cultivates, culture temperature is 35-40 DEG C, incubation time 1-2d;
2) slant culture: the Saccharomyces Cerevisiae in S C-X234 that step 1) middle plateform is cultivated is transferred on inclined-plane minimum medium, incubation time 18-20h, culture temperature 35-40 DEG C;
3) plant liquid to cultivate: by step 2) in the Saccharomyces Cerevisiae in S C-X234 of slant culture receive in seed culture medium and cultivate, incubation time 8-10h, culture temperature 35-40 DEG C, shaking speed is 150r/min;
4) fermentation culture: be transferred in fermention medium by the kind liquid in step 3), inoculum size is 10%, and incubation time is 48h, and culture temperature is 35-40 DEG C, and shaking speed is 100r/min.
Minimum medium wherein: step 1) and 2) comprises the component of following mass percent: carbon source 2%, nitrogenous source 3-4%, agar 2%, and all the other are water, pH nature; Wherein said carbon source is one or both in glucose or W-Gum, and described nitrogenous source is the mixture of peptone or yeast powder.
Step 3) seed culture medium comprises the component of following mass percent: carbon source 5%, nitrogenous source 0.5-1%, inorganic salt 0.01-0.02%, buffer reagent 0.1-0.2%, pH=5.5, and wherein carbon source is one or more mixtures in glucose, W-Gum or molasses; Nitrogenous source is one or more in yeast powder, ammonium sulfate or Secondary ammonium phosphate, and buffer reagent is primary ammonium phosphate.
Step 4) fermention medium comprises the component of following mass percent: carbon source 32-36%, nitrogenous source 1-2%, inorganic salt 0.02-0.05%, buffer reagent 0.5-1%, pH=5.5, and wherein carbon source is one or more mixtures in glucose or W-Gum; Nitrogenous source is one or more in yeast extract paste, ammonium sulfate, and buffer reagent is primary ammonium phosphate.
Injecting mediation spt15 gene is to improve the patience of yeast saccharomyces cerevisiae to ethanol, spt15 is hansen initiation transcription factor, the sudden change of spt15 gene can make the alcohol tolerance of yeast saccharomyces cerevisiae change, and in order to screen the yeast saccharomyces cerevisiae of high resistance to ethanol, thus obtains the yeast saccharomyces cerevisiae of high-yield ethanol.
Beneficial effect: present invention employs low energy N
+inject mediation spt15 gene engineering, with SC-100 for the bacterium that sets out, utilize that TTC is dull and stereotyped, alcohol is dull and stereotyped carries out primary dcreening operation, the bacterial strain obtained carries out the multiple sieve that ferments, improve the patience of yeast in alcohol gradually, final acquisition output can reach the production bacterial strain SC-X234 of 15.75%, and this bacterial strain can improve the low problem of fermentation production rate greatly, and sugar alcohol transformation efficiency is higher.In 5L fermentor tank, ethanol production can reach 126g/L, improves 87.5%, have very high economic worth as alcohol fuel than the original bacterium that sets out.
Embodiment
According to following embodiment, the present invention may be better understood.But those skilled in the art will readily understand, concrete material proportion, processing condition and result thereof described by embodiment only for illustration of the present invention, and should can not limit the present invention described in detail in claims yet.
The screening method of embodiment 1 Saccharomyces Cerevisiae in S C-X234
Starting strain Saccharomyces Cerevisiae in S C-100 is convenient to mark purchased from the Angel Yeast dry powder in Angel Yeast stock company, and laboratory is by its called after SC-100.
With SC-100 for the bacterium screening Saccharomyces Cerevisiae in S C-X234 that sets out, concrete steps are as follows:
A) preparation of monospore suspension: the bacterium that set out by yeast saccharomyces cerevisiae makes spore suspension, concentration is 1 × 10
6individual/mL.
B) mutagenesis of Mediated by Low Energy Ion Beam spt15 gene: get 100 μ L spore suspensions and the 50 μ L plasmid containing spt15, even spread, on aseptic flat board, dries up with sterile wind, under 20Kev energy, 40 × 10
14ions/cm
2carry out Nitrogen ion mutagenesis under dosage, after ion implantation, carry out wash-out in an aseptic environment with 1mLTE solution, the bacterium liquid after wash-out, at 37 DEG C of temperature bath 1h, is then applied on alcohol flat board, carries out pressure screening, be inverted for 37 DEG C and cultivate 2-3d.
C) screening of mutagenic fungi
The primary dcreening operation of TTC flat board: be poured on by TTC upper strata substratum low temperature on the middle flat board grown of step (b), be inverted at 37 DEG C and cultivate 1-2d, screen the bacterial strain that cell viability is strong, dehydrogenase activity is stronger, and redness is darker.
D) the multiple sieve of fermentation: in S. cervisiae access slant medium step (c) filtered out, cultivate 1-2d at 37 DEG C, then inclined-plane proceeds in kind of liquid, cultivates 8-10h at 37 DEG C.Get in seed liquor access fermention medium, inoculum size 10%(v/v), 250mL shaking flask dress 30mL substratum, rotating speed is 100r/min, leavening temperature 37 DEG C, fermentation 48h, measures the content of alcohol in fermented liquid, filter out the starting strain of the highest S. cervisiae of ethanol content as next round mutagenesis screening, repeat above-mentioned steps until screen aimed strain SC-X234.
Wherein culture medium prescription is:
Step b) in alcohol plate culture medium be: glucose 2%, fish meal protein peptone 2%, agar 2%, yeast extract paste 2%, alcohol 8% all the other for water pH nature.Step c) in TTC substratum be: TTC0.5g, glucose 5g, agar 15g constant volume to 1L nature pH
The TTC flat board of step (c) is divided into upper strata substratum and lower floor's substratum, and lower floor's substratum is YPD substratum, and upper strata nutrient media components is that TTC0.5g glucose 5g agar 15g constant volume is to 1L, pH nature.
Seed liquor substratum in step d): glucose 5%, yeast extract paste 0.5%, magnesium sulfate 0.01%, calcium chloride 0.06g/L, ammonium sulfate 0.2%, Secondary ammonium phosphate 0.1%, pH are 5.5, wherein glucose needs to divide disappear (point meaning disappeared for separating independent sterilizing).
Fermention medium in step d): glucose 36%, yeast extract paste 0.1%, calcium chloride 0.01%, ammonium sulfate 1%, Secondary ammonium phosphate 0.5%, magnesium sulfate 0.01%, pH5.5, wherein glucose needs point disappear.
Fermentation results ethanol content detects as table 1
Table 1
| Bacterium number | Set out bacterium SC-100 | Mutagenic fungi SC-X234 |
| Ethanol content (v/v) | 8% | 15% |
Have greatly improved through the ability of the product alcohol of the yeast of the mutagenic and breeding bacterium that comparatively sets out.
Embodiment 2
The present embodiment illustrates biology morphology and the genetic stability of mutagenic fungi Saccharomyces Cerevisiae in S C-X234
The morphology of Saccharomyces Cerevisiae in S C-X234 of the present invention and physiochemical properties: bacterium colony is creamy white, amphimicrobian, bacterium colony size is 2-3mm, circular colonies, growth temperature 35-40 DEG C, and optimal pH is 5.0-5.5, and methylene blue dyes: viable bacteria is colourless, and dead bacterium is blue.
Carry out passage assays with the substratum in embodiment 1 and culture condition, result as table 2,
The genetic stability of table 2 Saccharomyces Cerevisiae in S C-X234
| Passage number | Alcohol output (v/v) |
| 1 | 15.25 |
| 2 | 15 |
| 3 | 15 |
| 4 | 15.25 |
| 5 | 14.75 |
| 6 | 15.25 |
From genetic stability interpretation, mutant strain Saccharomyces Cerevisiae in S C-X234 is through 6 passage assays, and its alcohol output is stablized, and has good genetic stability, can as the bacterial strain of further R and D.
Embodiment 3
This example illustrates that Saccharomyces Cerevisiae in S C-X234 utilizes corn powder saccharification liquid to ferment and produces alcohol
Described in the present embodiment, culture medium prescription is as follows:
Plate culture medium: glucose 2%, peptone 2%, agar 2%, yeast extract paste 2%, all the other are water, natural pH.
Slant medium: glucose 2%, peptone 2%, agar 2%, yeast extract paste 2% all the other be water, natural pH.
Seed culture medium: glucose 5%, yeast extract paste 0.5%, magnesium sulfate 0.01%, calcium chloride %, ammonium sulfate 0.2%, Secondary ammonium phosphate 0.1%, pH are 5.5, wherein glucose needs point disappear.
Fermention medium: corn powder saccharification liquid 32%, yeast extract paste 0.1%, calcium chloride 0.01%, ammonium sulfate 1%, Secondary ammonium phosphate 0.5%, magnesium sulfate 0.01%pH5.5, wherein glucose needs point disappear, and corn powder saccharification liquid is the product of corn after α-amylase and saccharifying enzyme process.
Saccharomyces Cerevisiae in S C-X234 is inoculated into plate culture medium, culture temperature 37 DEG C, incubation time 12h-24h, then proceeds to slant medium, at 37 DEG C, cultivate 1-2d, and then inclined-plane proceeds in seed culture medium, at 37 DEG C, cultivate 8-10h.Get in seed liquor access fermention medium, inoculum size 10%(v/v), 250mL shaking flask dress 30mL substratum, rotating speed is 100r/min, leavening temperature 37 DEG C, fermentation 48h, the content measuring alcohol in fermented liquid reaches 13.5%(v/v), improve 50.6% than the original bacterium that sets out of equal conditions.
Embodiment 4
This example illustrates that mutant strain SC-X234 yeast saccharomyces cerevisiae utilizes molasses fermented product alcohol
This tests described assay medium formula
Plate culture medium: glucose 2%, fish meal protein peptone 2%, agar 2%, yeast extract paste 2%, all the other are water, natural pH.
Slant medium: glucose 2%, fish meal protein peptone 2%, agar 2%, yeast extract paste 2%, all the other are water, natural pH.
Seed culture medium: glucose 5%, yeast extract paste 0.5%, magnesium sulfate 0.01%, calcium chloride 0.06%, ammonium sulfate 0.2%, Secondary ammonium phosphate 0.1%, pH are 5.5, wherein glucose needs point disappear.
Fermention medium: molasses 32%, yeast extract paste 0.1%, calcium chloride 0.01%, ammonium sulfate 1%, Secondary ammonium phosphate 0.5%, magnesium sulfate 0.01%, pH5.5, wherein glucose needs point disappear.
Saccharomyces Cerevisiae in S C-X234 is inoculated into plate culture medium, culture temperature 37 DEG C, incubation time 12h-24h, then proceeds to slant medium, at 37 DEG C, cultivate 1-2d, and then inclined-plane proceeds in kind of liquid, at 37 DEG C, cultivate 8-10h.Get in seed liquor access fermention medium, inoculum size 10%(v/v), 250mL shaking flask dress 30mL substratum, rotating speed is 100r/min, leavening temperature 37 DEG C, fermentation 48h, the content measuring alcohol in fermented liquid reaches 10.5%(v/v), improve 75% than the original bacterium that sets out of equal conditions.
Embodiment 5
This example illustrates that mutant strain SC-X234 yeast saccharomyces cerevisiae produces alcohol in 5L fermentation cylinder for fermentation
This tests described assay medium formula
Plate culture medium: glucose 2%, fish meal protein peptone 2%, agar 2%, yeast extract paste 2% all the other be water, natural pH.
Slant medium: glucose 2%, fish meal protein peptone 2%, agar 2%, yeast extract paste 2% all the other be water, natural pH.
Seed culture medium: glucose 5%, yeast extract paste 0.5%, magnesium sulfate 0.01%, calcium chloride 0.06%, ammonium sulfate 0.2%, Secondary ammonium phosphate 0.1%, pH are 5.5, wherein glucose needs point disappear.
Fermention medium: glucose 32%, yeast extract paste 0.1%, calcium chloride 0.01%, ammonium sulfate 1%, Secondary ammonium phosphate 0.5%, magnesium sulfate 0.01%, pH5.5, wherein glucose needs point disappear.
Saccharomyces Cerevisiae in S C-X234 is inoculated into plate culture medium, culture temperature 37 DEG C, incubation time 12h-24h, then proceeds to slant medium, at 37 DEG C, cultivate 1-2d, and then inclined-plane proceeds in kind of liquid, at 37 DEG C, cultivate 8-10h.Get seed liquor access containing in the 5L fermentor tank of 3L fermention medium, inoculum size 10%(v/v), rotating speed is 100r/min, leavening temperature 37 DEG C, fermentation 48h, and detect ethanol content and reach 15.75%, the starting strain on year-on-year basis under the same terms improves 96.9%.
In above embodiment, alcohol detection method is: chromatography of gases (capillary column AC-20) post case temperature: 60 DEG C; Sampler temperature: 190 DEG C; Detector temperature: 240 DEG C.Internal standard substance: n-propyl alcohol.Detect liquid collocation method: (0.5mL6% n-propyl alcohol solution+0.5mL sample) is settled to 10mL.Sample size: 2 μ L.
In culture medium prescription of the present invention, per-cent (%) is mass percent.
Claims (6)
1. a yeast saccharomyces cerevisiae (
saccharomycescerevisiae) SC-X234, be preserved in China typical culture collection center CCTCC, deposit number: CCTCCM2014005.
2. the application of Saccharomyces Cerevisiae in S C-X234 according to claim 1 in fermentative production of ethanol.
3. the application of Saccharomyces Cerevisiae in S C-X234 according to claim 2 in fermentative production of ethanol, is characterized in that comprising the steps:
1) slat chain conveyor: Saccharomyces Cerevisiae in S C-X234 is connected on dull and stereotyped minimum medium and cultivates, culture temperature is 35-40 DEG C, incubation time 1-2d;
2) slant culture: the Saccharomyces Cerevisiae in S C-X234 that step 1) middle plateform is cultivated is transferred on inclined-plane minimum medium, incubation time 18-20h, culture temperature 35-40 DEG C;
3) plant liquid to cultivate: by step 2) in the Saccharomyces Cerevisiae in S C-X234 of slant culture receive in seed culture medium and cultivate, incubation time 8-10h, culture temperature 35-40 DEG C;
4) fermentation culture: be transferred in fermention medium by the Saccharomyces Cerevisiae in S C-X234 planting liquid cultivation in step 3), inoculum size is 10%, and incubation time is 48h, and culture temperature is 35-40 DEG C.
4. the application of Saccharomyces Cerevisiae in S C-X234 according to claim 3 in fermentative production of ethanol, it is characterized in that: step 1) and 2) in minimum medium comprise the component of following mass percent: carbon source 2%, nitrogenous source 3-4%, agar 2%, all the other are water, pH nature; Wherein said carbon source is one or both in glucose or W-Gum, and described nitrogenous source is peptone or yeast extract paste.
5. the application of Saccharomyces Cerevisiae in S C-X234 according to claim 3 in fermentative production of ethanol, it is characterized in that: step 3) seed culture medium comprises the component of following mass percent: carbon source 5%, nitrogenous source 0.5-1%, inorganic salt 0.01-0.02%, buffer reagent 0.1-0.2%, pH=5.5, wherein carbon source is one or more mixtures in glucose, W-Gum or molasses; Nitrogenous source is one or more in yeast extract paste, ammonium sulfate or Secondary ammonium phosphate; Inorganic salt are one or more in magnesium sulfate and calcium chloride; Buffer reagent is primary ammonium phosphate.
6. the application of Saccharomyces Cerevisiae in S C-X234 according to claim 3 in fermentative production of ethanol, it is characterized in that: step 4) fermention medium comprises the component of following mass percent: carbon source 32-36%, nitrogenous source 1-2%, inorganic salt 0.02-0.05%, buffer reagent 0.5-1%, pH=5.5, wherein carbon source is one or more mixtures in glucose, molasses or W-Gum; Nitrogenous source is one or more in yeast extract paste, ammonium sulfate, and buffer reagent is primary ammonium phosphate, and inorganic salt are one or more in magnesium sulfate and calcium chloride.
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| CN104263665B (en) * | 2014-09-23 | 2017-10-10 | 南京工业大学 | One plant of lignin tolerance S. cervisiae and its application in bio-ethanol production |
| CN104277990B (en) * | 2014-09-25 | 2017-07-18 | 南京工业大学 | One Accharomyces cerevisiae and its application in producing fuel ethyl alcohol by ferment |
| CN105907779B (en) * | 2016-05-25 | 2019-07-09 | 西北农林科技大学 | A kind of recombinant plasmid pY16TEF1- △ SPT15 reducing Ethanol in Saccharomyces cerevisiae yield and its application |
| CN105950649B (en) * | 2016-05-25 | 2019-07-09 | 西北农林科技大学 | A kind of recombinant plasmid pY16TEF1- △ SPT15-125 that Ethanol in Saccharomyces cerevisiae yield can be improved and its application |
| CN109666705A (en) * | 2019-01-07 | 2019-04-23 | 山东理工大学 | Chromatin remodeling factors gene is in the application for improving fermentation alcohol yield in S. cervisiae |
| CN111040957A (en) * | 2019-09-30 | 2020-04-21 | 中国石油化工股份有限公司 | Ethanol-resistant and high-sugar-resistant fermentation strain and construction method thereof |
| CN112921056B (en) * | 2021-04-08 | 2022-10-11 | 中溶科技股份有限公司 | Method for continuously producing cellulase and co-producing ethanol under control of inducer |
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