CN101555494A - Method for preparing fuel of ethanol from immobilized mixed strain fermented cellulosic hydrolysate - Google Patents

Method for preparing fuel of ethanol from immobilized mixed strain fermented cellulosic hydrolysate Download PDF

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Publication number
CN101555494A
CN101555494A CNA200810103619XA CN200810103619A CN101555494A CN 101555494 A CN101555494 A CN 101555494A CN A200810103619X A CNA200810103619X A CN A200810103619XA CN 200810103619 A CN200810103619 A CN 200810103619A CN 101555494 A CN101555494 A CN 101555494A
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immobilized
hydrolyzed solution
hydrolysate
fermentation
yeast
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魏刚
李艳华
魏云鹏
张雯
房彩琴
熊蓉春
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Beijing University of Chemical Technology
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Beijing University of Chemical Technology
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

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Abstract

The invention relates to a method for preparing fuel of ethanol from immobilized mixed strain fermented cellulosic hydrolysate, which is characterized by utilizing mixed strains of P. tannophilus and S. cerevisilae to graft into diluted acid hydrolysate and fermenting the mixed solution by a supplemented material batch fermentation mode. The method is realized by the following steps: (1) domestication of the P. tannophilus and the S. cerevisilae; (2) immobilization of the mixed strains by calcium alginate; (3) multiplication culture of the immobilized mixed strains; and (4) supplemented material batch fermentation of the immobilized mixed strains. The immobilized mixed strains can be fermented while using xylems and glucose in the hydrolysate for fermentation, thereby the fermentation period is shortened, the utilization ratio of the xylose and the glucose is improved and the ethanol concentration is increased; the immobilized mixed strains can be repeatedly used and have strong anti-hybrid bacteria capability; the supplemented material batch fermentation mode is applied to hydrolysate with cellulose raw material for producing the fuel of ethanol, thereby reducing inhibiting factors in the hydrolysate; and the method is beneficial to thoroughly eliminating the inhibition on substrates, diluting poisonous metabolite, reducing contaminated bacterial, controlling the fermentation time and preventing the generation of side products.

Description

The immobilized mixed strain fermented cellulosic hydrolyzed solution prepares the method for alcohol fuel
Technical field
The present invention relates to the method that a kind of immobilized mixed strain prepares alcohol fuel, the fermented by mixed bacterium ligno-cellulose hydrolysate that specifically utilizes immobilization to have a liking for tannin pipe capsule yeast (Pachysolen tannophilus) and yeast saccharomyces cerevisiae (Saccharomycescerevisiae) prepares the production of fuel ethanol method.
Background technology
Ethanol is a kind of important industrial raw material, is widely used in fields such as chemical industry, food, military project, daily-use chemical industry and medical and health, can also be as basic material, the fuel of energy industry.The world today is faced with the challenge of environment and development, requires to use the cry of white gasoline to grow to even greater heights.The further dehydration of ethanol added again forming the distortion alcohol fuel behind an amount of gasoline, ethanol is regarded as substituting and saving the optimum fuel of gasoline abroad, and as a kind of bioenergy, ethanol is expected to replace the fossil oil (oil and coal) that reduces day by day.
The main raw material of alcohol production has Mierocrystalline cellulose and hemicellulose, starch and saccharine material etc.Wherein Mierocrystalline cellulose and hemicellulose are organic substances the abundantest, cheap and that be easy to get.According to estimates, the annual Mierocrystalline cellulose waste residue that can produce about 1,700 hundred million tons on the earth, making full use of that hexose that cellulosic material produces and wood sugar ferment is the key of current alleviating energy crisis.
Immobilization technology is used widely in fields such as industrial production, environmental engineerings.By the pair cell immobilization, can effectively increase cell concn, improve productive rate.Prepare the method for alcohol about immobilized cell fermentation, existing in recent years patent is announced.U.S. Pat 4393136 utilizes immobilization zymomonas mobilis zymohexose to prepare ethanol; U.S. Pat 4350765 is fixed in yeast saccharomyces cerevisiae and prepares ethanol in the alginate calcium; In bilayer capsule, xylose isomerase turns to xylulose and the xylulose fermenting alcohol carries out synchronously with xylose isomerase and urase co-immobilization for U.S. Pat 5254468.The said fixing technology is to increasing cell concn, improving alcohol yied has better effects, but fixedly turns to the master with single cell, and single hexose and wood sugar can only ferment, and cellulosic hydrolysate mainly is made up of hexose and wood sugar, utilizes two kinds of sugared co-fermentation to prepare ethanol simultaneously and is only key.Simultaneously, the fermented bacterium fermentation capacity is strong inadequately, and the xylose utilization rate is low, fermentation period is long, alcohol yied is lower etc. also is the problem that waits to solve.
Summary of the invention
The purpose of this invention is to provide the method that a kind of ligno-cellulose hydrolysate that ferments prepares alcohol fuel.In order to make full use of hexose and the wood sugar in the cellulosic hydrolysate, adopt immobilized mixed strain producing fuel ethyl alcohol by ferment altogether, in order to solve the problem of immobilized mixed strain fermented existence, provide a kind of new wood-sugar fermentation bacterial classification and yeast saccharomyces cerevisiae co-immobilization of utilizing, the fermentation capacity that employing feed supplement batch fermentation mode is fermented and improved alcohol concn, shortening fermentation period, improves bacterial classification.The present invention includes following steps:
To have a liking for tannin pipe capsule yeast (P.tannophilus) and yeast saccharomyces cerevisiae (S.cerevisiae) is tamed simultaneously, the ratio of hydrolyzed solution all is hydrolyzed solution along with the increase of the domestication number of times ratio growth by 20-50% to taming substratum in the substratum.With having a liking in tannin pipe capsule yeast and the sodium alginate soln of yeast saccharomyces cerevisiae bacteria suspension that domestication is cultivated, inject 2%CaCl in 1: 1 ratio access 3% 2In the aqueous solution, solidify 4h, sterilized water washing sodium alginate gel ball (diameter 2-3mm) moves in 1% alum liquor, 0-4 ℃ of gel ball that solidifies being fixed of 24h mixed strains down.
Immobilized gel ball is put in the proliferated culture medium, 30 ℃, multiplication culture 20-48h under the 100r/min condition, after treating that propagation is good, the filling ratio of immobilization gel ball by 20-50% joined in the fermention medium of ligno-cellulose hydrolysate, initial sugared concentration is 3-15%, regulating the pH value is 3.0-7.0, shaker fermentation under 28-35 ℃, 100-150r/min condition replenishes fresh hydrolysis sugar liquid behind the 12-24h at interval, carries out fed-batch fermentation.
In above-mentioned preparation process:
Used yeast is for having a liking for tannin pipe capsule yeast (pachysolen tannophilus): Beijing University of Chemical Technology's preservation.Yeast saccharomyces cerevisiae (Saccharomyces cerevisiae): Beijing University of Chemical Technology's preservation.
When mixed strains was tamed, domestication was cultivated prescription and is: wood sugar 10-50g/L, glucose 10-50g/L, urea 2.5g/L, yeast extract paste 5g/L, (NH 4) 2SO 45g/L, KH 2PO 42g/L, MgSO 47H 2O 1g/L, hydrolyzed solution (20,40,60,70,80,100%), pH nature.30 ℃, 160rpm, each batch domestication time is 18-24h, inoculum size 10-20%.
When immobilized mixed strain was carried out multiplication culture, the multiplication culture based formulas was: wood sugar 10-50g/L, glucose 10-50g/L, yeast extract paste 5g/L, peptone 5g/L, KH 2PO 41g/L, MgSO 47H 2O 1g/L.
Carry out immobilized mixed strain and cellulosic hydrolysate is fermented fermentative medium formula: hydrolyzed solution concentrates liquid glucose 5%-15%, (NH 4) 2SO 40.5%, yeast extract paste 0.2%, KH 2PO 40.2%, MgSO 47H 2O0.1%.
Production alcohol fuel of the present invention can be rice straw, maize straw, corn cob, wood fragments bits, depleted Case Board, Corrugating Base Paper, depleted newspaper and office with paper etc. with fibrous material.
The present invention is used has a liking for the bacterial classification that tannin pipe capsule yeast is a kind of zymohexose simultaneously and wood sugar, tannin pipe capsule yeast is had a liking in immobilization after the utilization domestication is handled and the yeast saccharomyces cerevisiae mixed strains carries out the fermentation of dilute acid hydrolysis liquid, improved the fermentation capacity of bacterial classification, utilize wood sugar, glucose in the hydrolyzed solution to ferment simultaneously, shorten fermentation period, improved sugared utilization ratio, increased alcohol concn, immobilized mixed strains can be reused, and anti-assorted bacterium ability strengthens; Simultaneously feed supplement batch fermentation mode is applied to the cellulosic material hydrolyzed solution and prepares alcohol fuel, it is with respect to the advantage of other fermentation modes: the one, and the supressor in the hydrolyzed solution is joined in the reactive system by a small amount of batch-wise, can avoid once a large amount of the adding and the bacterial metabolism that causes is subjected to the influence of environment flip-flop; The 2nd, the feed supplement batch fermentation helps eliminating substrate and suppresses, dilutes the toxic metabolite product, reduces microbiological contamination and control fermentation time.
Embodiment
Embodiment 1:
What domestication was cultivated has a liking in tannin pipe capsule yeast and the sodium alginate soln of yeast saccharomyces cerevisiae bacteria suspension in 1: 1 ratio access 3%, injects 2%CaCl 2In the aqueous solution, solidify 4h, sterilized water washing sodium alginate gel ball (diameter 2-3mm) moves in 1% alum liquor, 0-4 ℃ of gel ball that solidifies being fixed of 12-24h mixed strains down.Immobilized gel ball is put in the proliferated culture medium, 30 ℃, multiplication culture 48h under the 100r/min condition, treat that propagation is good after, the immobilization gel ball is moved in the fermention medium of ligno-cellulose hydrolysate, initial sugared concentration 143.41g/L, regulating the pH value is 5.6, the stacking volume of fermentating liquid volume and gel ball is 3: 2,30 ℃, the constant temperature shaking table top fermentation of 100r/min, fermentation 12h, the mensuration ethanol concn is 11.42g/L, spirit yield is 53.6%.
Embodiment 2:
Other is with embodiment 1, the immobilized mixed strain that propagation is good moves in the cellulosic hydrolysate fermention medium, initial sugared concentration 88.428g/L, regulating the pH value is 5.7, and the stacking volume of fermentating liquid volume and gel ball is 3: 2,30 ℃, the constant temperature shaking table top fermentation of 100r/min, replenish fresh hydrolyzed solution behind the 12h at interval, fermentation 24h, the mensuration ethanol concn is 15.09g/L, spirit yield is 78.5%.
Embodiment 3:
Other is with embodiment 1, the immobilized mixed strain that propagation is good moves in the cellulosic hydrolysate fermention medium, initial sugared concentration 118.42g/L, regulating the pH value is 6.0, and the stacking volume of fermentating liquid volume and gel ball is 3: 2,32 ℃, the constant temperature shaking table top fermentation of 110r/min, replenish fresh hydrolyzed solution behind the 12h at interval, fermentation 24h, the mensuration ethanol concn is 19.84g/L, spirit yield is 83.13%.
Embodiment 4:
Other is with embodiment 1, the immobilized mixed strain that propagation is good moves in the cellulosic hydrolysate fermention medium, initial sugared concentration 50.587g/L, regulating the pH value is 5.7, and the stacking volume of fermentating liquid volume and gel ball is 3: 2,32 ℃, the constant temperature shaking table top fermentation of 100r/min, replenish fresh hydrolyzed solution behind the 12h at interval, fermentation 24h, the mensuration ethanol concn is 6.47g/L, spirit yield is 35.05%.
Embodiment 5:
Other is with embodiment 1, the immobilized mixed strain that propagation is good moves in the cellulosic hydrolysate fermention medium, initial sugared concentration 148.908g/L, regulating the pH value is 5.2, and the stacking volume of fermentating liquid volume and gel ball is 3: 2,28 ℃, the constant temperature shaking table top fermentation of 120r/min, replenish fresh hydrolyzed solution behind the 24h at interval, fermentation 48h, the mensuration ethanol concn is 14.39g/L, spirit yield is 58.39%.
Embodiment 6:
Other is with embodiment 1, the immobilized mixed strain that propagation is good moves in the cellulosic hydrolysate fermention medium, initial sugared concentration 98.92g/L, regulating the pH value is 3.7, and the stacking volume of fermentating liquid volume and gel ball is 3: 2,33 ℃, the constant temperature shaking table top fermentation of 130r/min, replenish fresh hydrolyzed solution behind the 12h at interval, fermentation 24h, the mensuration ethanol concn is 11.421g/L, spirit yield is 53.6%.
Embodiment 7:
Other is with embodiment 1, the immobilized mixed strain that propagation is good moves in the cellulosic hydrolysate fermention medium, initial sugared concentration 32.97g/L, regulating the pH value is 5.6, and the stacking volume of fermentating liquid volume and gel ball is 3: 2,29 ℃, the constant temperature shaking table top fermentation of 100r/min, replenish fresh hydrolyzed solution behind the 24h at interval, fermentation 70h, the mensuration ethanol concn is 6.67g/L, spirit yield is 37.72%.
Embodiment 8:
Other is with embodiment 1, the immobilized mixed strain that propagation is good moves in the cellulosic hydrolysate fermention medium, initial sugared concentration 88.428g/L, regulating the pH value is 5.2, and the stacking volume of fermentating liquid volume and gel ball is 3: 2,33 ℃, the constant temperature shaking table top fermentation of 100r/min, replenish fresh hydrolyzed solution behind the 12h at interval, fermentation 24h, the mensuration ethanol concn is 12.412g/L, spirit yield is 56.05%.

Claims (6)

1, a kind of immobilized mixed strain fermented cellulosic hydrolyzed solution prepares the method for alcohol fuel, it is characterized in that the mixed strains that utilizes immobilization to have a liking for tannin pipe capsule yeast (Pachysolen tannophilus) and yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) inserts dilute acid hydrolysis liquid, adopts the mode of feed supplement batch fermentation to ferment.Produce alcohol fuel by following steps:
To have a liking for tannin pipe capsule yeast (P.tannophilus) and yeast saccharomyces cerevisiae (S.cerevisiae) is tamed simultaneously, the ratio of hydrolyzed solution all is hydrolyzed solution along with the increase of the domestication number of times ratio growth by 20-50% to taming substratum in the substratum.With having a liking in tannin pipe capsule yeast and the sodium alginate soln of yeast saccharomyces cerevisiae bacteria suspension that domestication is cultivated, inject 2%CaCl in 1: 1 ratio access 3% 2In the aqueous solution, solidify 2-6h, sterilized water washing sodium alginate gel ball (diameter 2-3mm) moves in 1% alum liquor, 0-4 ℃ of gel ball that solidifies being fixed of 12-24h mixed strains down.
Immobilized gel ball is put in the proliferated culture medium, 28-35 ℃, multiplication culture 20-48h under the 100-150r/min condition, after treating that propagation is good, the filling ratio of immobilization gel ball by 20-50% joined in the fermention medium of ligno-cellulose hydrolysate, initial sugared concentration is 3-15%, regulating the pH value is 3.0-7.0, shaker fermentation under 28-35 ℃, 100-150r/min condition replenishes fresh hydrolysis sugar liquid behind the 12-48h at interval, carries out fed-batch fermentation.
Above-mentioned domestication culture medium prescription is wood sugar 10-50g/L, glucose 10-50g/L, urea 2.5g/L, yeast extract paste 5g/L, (NH 4) 2SO 45g/L, KH 2PO 42g/L, MgSO 47H 2O 1g/L, 30 ℃, 160rpm, the hydrolyzed solution ratio increases progressively by 5-40%, and each batch domestication time is 18-24h, domestication batch 6-24 time, yeast liquid inoculum size 10-20%.
2, the immobilized mixed strain fermented cellulosic hydrolyzed solution prepares the method for alcohol fuel according to claim 1, it is characterized in that fermented bacterium is selected for use to have a liking for tannin pipe capsule yeast (P.tannophilus) and yeast saccharomyces cerevisiae (S.cerevisiae).
3, the immobilized mixed strain fermented cellulosic hydrolyzed solution prepares the method for alcohol fuel according to claim 1, it is characterized in that utilizing fixedly mixed strains of calcium alginate gel ball, add 1% Tai-Ace S 150 and strengthen the intensity of immobilization gel ball, be 12-24h set time.
4, the immobilized mixed strain fermented cellulosic hydrolyzed solution prepares the method for alcohol fuel according to claim 1, the ratio that it is characterized in that taming hydrolyzed solution in the substratum is by increasing progressively by 5-20%, each batch domestication time is 18-24h, domestication batch 6-24 time, yeast liquid inoculum size 10-20%.
5, the immobilized mixed strain fermented cellulosic hydrolyzed solution prepares the method for alcohol fuel according to claim 1, it is characterized in that the immobilization gel ball joins in the fermention medium of cellulosic hydrolysate by the filling ratio of 20-50%, the initial sugared concentration of cellulosic hydrolysate is 3-15%, the pH value is 3.0-7.0, shaker fermentation under 28-35 ℃, 100-150r/min condition.
6, the immobilized mixed strain fermented cellulosic hydrolyzed solution prepares the method for alcohol fuel according to claim 1, and it is characterized in that producing alcohol fuel can be maize straw, corn cob, rice straw, depleted Case Board, Corrugating Base Paper, disregard message paper, depleted office with paper etc. with raw material.
CNA200810103619XA 2008-04-09 2008-04-09 Method for preparing fuel of ethanol from immobilized mixed strain fermented cellulosic hydrolysate Pending CN101555494A (en)

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Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101851656A (en) * 2010-04-19 2010-10-06 中国科学院青岛生物能源与过程研究所 Method for producing cellulosic ethanol
CN101906489A (en) * 2010-07-28 2010-12-08 张琴 Method for dilute acid hydrolysis of cotton stalks and method for producing alcohol
CN102093961A (en) * 2010-12-13 2011-06-15 山东轻工业学院 Brewing yeast SC1230 and application thereof in producing ethanol
CN102154382A (en) * 2011-01-11 2011-08-17 南阳天冠生物发酵有限公司 Method for producing cellulosic ethanol by using 1308 Saccharomyces cerevisiae
CN102635871A (en) * 2012-04-18 2012-08-15 东方电气集团东方锅炉股份有限公司 Method and device for solving contamination of boiler convection heating surface
CN103898166A (en) * 2012-12-25 2014-07-02 中粮营养健康研究院有限公司 Method of producing ethanol
CN103923905A (en) * 2013-01-16 2014-07-16 浙江海洋学院 Preparation method for immobilized microbe oil-spill repairing agent
CN104046658A (en) * 2014-06-26 2014-09-17 无锡市崇安区科技创业服务中心 Alcohol fermentation culture medium and application thereof
CN105296329A (en) * 2015-11-09 2016-02-03 北京化工大学 Device and method for performing continuous pretreatment on lignocellulose material by utilizing microwave-diluted acid steam explosion
CN114107161A (en) * 2021-11-15 2022-03-01 泸州老窖股份有限公司 Method for producing ellagic acid by degrading pomegranate rind with mixed domesticated strain

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101851656B (en) * 2010-04-19 2012-08-29 中国科学院青岛生物能源与过程研究所 Method for producing cellulosic ethanol
CN101851656A (en) * 2010-04-19 2010-10-06 中国科学院青岛生物能源与过程研究所 Method for producing cellulosic ethanol
CN101906489A (en) * 2010-07-28 2010-12-08 张琴 Method for dilute acid hydrolysis of cotton stalks and method for producing alcohol
CN102093961A (en) * 2010-12-13 2011-06-15 山东轻工业学院 Brewing yeast SC1230 and application thereof in producing ethanol
CN102093961B (en) * 2010-12-13 2012-09-05 山东海瑞特生物工程有限公司 Brewing yeast SC1230 and application thereof in producing ethanol
CN102154382A (en) * 2011-01-11 2011-08-17 南阳天冠生物发酵有限公司 Method for producing cellulosic ethanol by using 1308 Saccharomyces cerevisiae
CN102154382B (en) * 2011-01-11 2012-08-01 南阳天冠生物发酵有限公司 Method for producing cellulosic ethanol by using 1308 Saccharomyces cerevisiae
CN102635871A (en) * 2012-04-18 2012-08-15 东方电气集团东方锅炉股份有限公司 Method and device for solving contamination of boiler convection heating surface
CN102635871B (en) * 2012-04-18 2014-08-27 东方电气集团东方锅炉股份有限公司 Method and device for solving contamination of boiler convection heating surface
CN103898166A (en) * 2012-12-25 2014-07-02 中粮营养健康研究院有限公司 Method of producing ethanol
CN103923905A (en) * 2013-01-16 2014-07-16 浙江海洋学院 Preparation method for immobilized microbe oil-spill repairing agent
CN104046658A (en) * 2014-06-26 2014-09-17 无锡市崇安区科技创业服务中心 Alcohol fermentation culture medium and application thereof
CN105296329A (en) * 2015-11-09 2016-02-03 北京化工大学 Device and method for performing continuous pretreatment on lignocellulose material by utilizing microwave-diluted acid steam explosion
CN105296329B (en) * 2015-11-09 2018-05-25 北京化工大学 A kind of device and method continuously pre-processed to lignocellulose material using microwave-diluted acid steam explosion
CN114107161A (en) * 2021-11-15 2022-03-01 泸州老窖股份有限公司 Method for producing ellagic acid by degrading pomegranate rind with mixed domesticated strain

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