CN100339483C - Process for preparing alcohol through utilizing xylose and glucose by microzyme - Google Patents
Process for preparing alcohol through utilizing xylose and glucose by microzyme Download PDFInfo
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- CN100339483C CN100339483C CNB2003101218299A CN200310121829A CN100339483C CN 100339483 C CN100339483 C CN 100339483C CN B2003101218299 A CNB2003101218299 A CN B2003101218299A CN 200310121829 A CN200310121829 A CN 200310121829A CN 100339483 C CN100339483 C CN 100339483C
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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
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- 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 present invention provides a method for producing alcohol by xylose and glucose, which relates to the application of microzyme, particularly to a method for producing the alcohol by using one of tannophil pachysolen tannophilus boidin et adzet 2.1662, tannophil pachysolen tannophilus boidin et adzet ATCC32691 and candida shehatae buckle. Et v. uden ATCC34887 and using the xylose and the glucose as substrates. The present invention uses technical measures such as acclimatization, fluid culture, immobilization, propagation culture, etc. on the basis of the existing microzyme strains, so the period for fermenting and producing the alcohol by the microzyme is shortened, and the substrate utilization ratio is improved. The method of the present invention is mainly used for fermenting and producing the alcohol by using biomass as raw materials.
Description
Technical field
The present invention relates to saccharomycetic application, particularly relate to the method that one of three saccharomycete kind pachysolen tannophilus (Pachysolen tannophilus) 2.1622, pachysolen tannophilus ATCC 32691, shehatae candida (Candida shehatae) ATCC 34887 utilize wood sugar and glucose production alcohol.
Background technology
Alcohol claims ethanol again, is fermenting substrate production with the hexose by yeast saccharomyces cerevisiae usually.With biomass (agricultural crop straw, wood working refuse etc.) is that the raw material production alcohol fuel is the important component part of national energy strategy and national energy security, also being to prevent and reduce automobile burning tail gas and cause the air-polluting important channel, is the important assurance of national economy and social sustainable development.In biomass, Mierocrystalline cellulose and hemicellulose are main chemical compositions, and hemicellulose comprises various poly-pentoses and hexosan, and modal hemicellulose is an xylan, and it accounts for half of herbaceous plant dry weight, also is present in the xylophyta.Lignocellulose can mainly be contained the hydrolysis sugar liquid of glucose and wood sugar after pre-treatment.Is that raw material carries out in the method for zymamsis in research with biomass, and key is will determine and can become the bacterial strain of alcohol with glucose by simultaneously efficient metabolism wood sugar, and then improves technology of alcohol on this basis.The wood sugar of metabolism simultaneously becomes the yeast of alcohol that pachysolen tannophilus is arranged with glucose now, shehatae candida, pichia spp (Pichia stipitis), kluyveromyces marxianus (Kluyveromyces marxianus) and candiyeast (Candida sp XF217) etc., but the actual output that they utilize wood sugar and glucose fermentation to produce alcohol has only about 70% of theoretical yield, and fermentation period is long, about 80 hours.
Summary of the invention
The purpose of this invention is to provide the method that a kind of yeast utilizes wood sugar and glucose fermentation to produce alcohol, this method is based on existing yeast strain, by adopting technique means such as domestication, liquid culture, immobilization, the zymamsis cycle is shortened, substrate utilization ratio improves.
For realizing purpose of the present invention, adopted following technical scheme, this method may further comprise the steps:
1) pachysolen tannophilus 2.1622 being inoculated in the domestication substratum tames;
2) yeast after will taming is inoculated in and carries out liquid culture in the liquid nutrient medium;
3) yeast that liquid culture is obtained carries out immobilization;
4) immobilized yeast is carried out multiplication culture;
5) yeast after the propagation carries out zymamsis;
In the method for above-mentioned production alcohol:
When bacterial strain was tamed, the prescription of domestication substratum was: wood sugar 10-50g/L, peptone 4-6g/L, yeast water 2-4g/L, the domestication condition is pH5.0-5.5,28-35 ℃, and 60-120rpm, each batch domestication time is 48-96 hour, and inoculum size is 2%, and domestication is more than 5 batches.
When the yeast after the domestication is carried out liquid culture, the prescription of liquid nutrient medium is: wood sugar 10-50g/L, peptone 4-6g/L, yeast water 2-4g/L, inoculum size is 1-5%, culture condition is pH5.0-5.5,28-35 ℃, 60-120rpm, incubation time are 24-48 hour, cultivate and finish the back concentration, it is standby to collect nutrient solution.
When the yeast that liquid culture is obtained carries out immobilization, at every turn with the concentrated liquid culture of 2ml with contain 1.2%Al
2O
3The 3-6% sodium alginate soln mix at normal temperatures, after under aseptic condition, stirring, be added drop-wise to 2% CaCl
2In the aqueous solution, after leaving standstill 10-20 hour under 4 ℃, obtain gel particles.
When fixed yeast cell was carried out multiplication culture, the multiplication culture based formulas was: wood sugar 20g/L, glucose 50g/L, peptone 3g/L, yeast water 2.5g/L, CaCl
20.25g/L, MgSO
40.25g/L, KH
2PO
42.5g/L the multiplication culture condition is 28-35 ℃, pH5.0-5.5, and 60-120rpm, incubation time were 24-48 hour, changed a fresh medium every 12 hours.
Utilize wood sugar and glucose to carry out zymamsis at last, the prescription of fermention medium is: wood sugar 20g/L, glucose 50g/L, CaCl
22.5g/L, MgSO
40.25g/L, KH
2PO
42.5g/L, yeast water 2.5g/L, peptone 3g/L, urea 0.24g/L, fermentation condition are pH5.0-5.5,28-35 ℃, 60-120rmp, fermentation time are 24-36 hour.
Use high-pressure liquid phase chromatograph measuring in the laboratory:
The xylose utilization rate: the immobilized cell after pachysolen tannophilus 2.1622, pachysolen tannophilus ATCC 32691 and the domestication of shehatae candida ATCC 34,887 three strain bacterial strains is respectively 87.90% to the utilization ratio of wood sugar, 76.62% and 74.68%, pachysolen tannophilus 2.1622 obviously is better than other two bacterial strains.
The glucose utilization rate: the immobilized cell after the above-mentioned three bacterial strains domestication also is significantly improved to the ability of utilizing of glucose.Before the domestication, immobilized cell fermentation 36h, the glucose utilization rate of pachysolen tannophilus 2.1622, pachysolen tannophilus ATCC 32691 and shehatae candida ATCC 34,887 three strain bacterial strains is respectively 99.21%, 96.91%, 92.01%.After the domestication, immobilized cell fermentation 24h, above-mentioned three bacterial strains have all reached more than 85% the utilization ratio of glucose.Fermentation 36h, the glucose utilization rate of pachysolen tannophilus 2.1622, pachysolen tannophilus ATCC 32691 and shehatae candida ATCC 34,887 three strain bacterial strains is respectively 99.99%, 98.21%, 97.46%.48h, the glucose utilization rate of above-mentioned three strain bacterium has all reached 99.99%, near 100%.For immobilized cell pachysolen tannophilus 2.1622, fermentation 24h, the glucose utilization rate has promptly reached 99.99%, and fermentation time has shortened 1/3rd.
The total reducing sugar utilization ratio: before the domestication, the total reducing sugar utilization ratio of pachysolen tannophilus 2.1622, pachysolen tannophilus ATCC 32691 and shehatae candida ATCC 34,887 three strain bacterial strain immobilized cells is respectively 85.08%, 79.85%, 77.81%.After the domestication, the total reducing sugar utilization ratio of above-mentioned three bacterial strain immobilized cells respectively is 93.95%, 88.31%, 87.34%.
The alcohol yied of domestication back three bacterial strains: the alcohol yied of domestication back three bacterial strains has obtained significantly improving.In general, in three bacterial strains, bacterial strain pachysolen tannophilus 2.1622 is better than other two bacterial strains.
Yeast of the present invention utilizes wood sugar and glucose to carry out zymamsis to have the advantages that the cycle is short, substrate conversion efficiency is high, alcohol output is high.
The invention will be further described below by specific embodiment.
Embodiment
Embodiment 1
Pachysolen tannophilus 2.1622 utilizes the method for wood sugar and glucose production alcohol.
The domestication substratum of 100ml is housed in the triangular flask of 250ml, and the prescription of domestication substratum is: wood sugar 20g/L, peptone 5g/L, yeast water 3g/L.With the yeast pachysolen tannophilus 2.1622 that the inclined-plane is preserved, be inoculated in the domestication substratum, inoculum size is 2%.At pH5.0,30 ℃, cultivated under the condition of 60rpm 48 hours, carry out 7 batches domestication altogether and cultivate.With the microscope direct-counting method bacteria containing amount in the nutrient solution is counted, bacteria containing amount increases and obviously increases with domestication batch, and pachysolen tannophilus 2.1622 is after cultivating through 7 batches domestication, and bacteria containing amount reaches 3.3 * 10
8/ ml, and the bacteria containing amount of this bacterial strain after the 1st domestication has only 5.0 * 10
7/ ml, bacteria containing amount has increased by 1 order of magnitude again on the basis after the domestication for the first time.The increase of bacteria containing amount illustrates that the ability of its metabolism wood sugar and glucose improves.
Yeast pachysolen tannophilus 2.1622 after the domestication is carried out liquid culture.The liquid nutrient medium of 100ml is housed in the triangular flask of 250ml, and the prescription of liquid nutrient medium is: wood sugar 20g/L, peptone 5g/L, yeast water 3g/L.Pachysolen tannophilus 2.1622 after the domestication is inoculated in the liquid nutrient medium, and inoculum size is 2%.At pH5.0,30 ℃, cultivated 24 hours under the condition of 60rpm, cultivate and finish the back concentration, it is standby to collect nutrient solution.
Use this culture as inoculum then, pachysolen tannophilus 2.1622 cells are carried out immobilization.Get the step cultivation bacterium liquid 2ml and the 68ml that obtain at every turn and contain 1.2%Al
2O
33% sodium alginate soln mix at normal temperatures, after under aseptic condition, stirring, be added drop-wise to 2% CaCl
2In the aqueous solution, under 4 ℃, left standstill 10 hours.Pachysolen tannophilus 2.1622 cells are made gel particles through after the immobilization, and its diameter D is about 3mm.After the immobilization gel particles usefulness sterile water wash three times, change in the proliferated culture medium.
Immobilized pachysolen tannophilus 2.1622 is carried out multiplication culture.The proliferated culture medium of 100ml is housed in the triangular flask of 250ml, and the prescription of proliferated culture medium is: wood sugar 20g/L, glucose 50g/L, peptone 3g/L, yeast water 2.5g/L, CaCl
20.25g/L, MgSO
40.25g/L, KH
2PO
42.5g/L.Gel particles is inserted in the proliferated culture medium of 100ml, at 30 ℃, pH5.0 cultivated 24 hours under the condition of 80rpm, will change one time fresh medium after 12 hours.
Carry out zymamsis with the pachysolen tannophilus behind the multiplication culture 2.1622 at last.The fermention medium of 100ml is housed in the triangular flask of 250ml, and the prescription of fermention medium is: wood sugar 20g/L, glucose 50g/L, CaCl
22.5g/L, MgSO
40.25g/L, KH
2PO
42.5g/L, yeast water 2.5g/L, peptone 3g/L, urea 0.24g/L.Pachysolen tannophilus behind the multiplication culture 2.1622 is inserted in the fermention medium of 100ml,, 30 ℃, carried out fermentation reaction under the condition of pH value 5.0 24 hours at 60rpm.
Measure through high pressure liquid chromatograph: the xylose utilization rate is 87.90%, and the glucose utilization rate has reached 99.99%, and the total reducing sugar utilization ratio is 93.95%.With the ethanol content in the gas chromatographic analysis fermented liquid, the presentation of results alcohol yied reaches 97%.
Embodiment 2
Pachysolen tannophilus ATCC 32691 utilizes the method for wood sugar and glucose production alcohol.
The domestication substratum of 100ml is housed in the triangular flask of 250ml, and the prescription of domestication substratum is: wood sugar 10g/L, peptone 4g/L, yeast water 2g/L.With the yeast pachysolen tannophilus ATCC32691 that the inclined-plane is preserved, be inoculated in the domestication substratum, inoculum size is 3%.At pH is 5.5,35 ℃, cultivates under the condition of 80rpm 60 hours, carries out 6 batches domestication altogether and cultivates.With the microscope direct-counting method bacteria containing amount in the nutrient solution is counted, bacteria containing amount increases and obviously increases with domestication batch.The increase of bacteria containing amount illustrates that the ability of its metabolism wood sugar and glucose improves.
Yeast pachysolen tannophilus ATCC 32691 after the domestication is carried out liquid culture.The liquid nutrient medium of 100ml is housed in the triangular flask of 250ml, and the prescription of liquid nutrient medium is: wood sugar 10g/L, peptone 4g/L, yeast water 2g/L.Pachysolen tannophilus ATCC 32691 after the domestication is inoculated in the liquid nutrient medium, and inoculum size is 3%.At pH5.5,35 ℃, cultivated 36 hours under the condition of 80rpm, cultivate and finish the back concentration, it is standby to collect nutrient solution.
Use this culture as inoculum then, pachysolen tannophilus ATCC 32691 cells are carried out immobilization.Get the step cultivation bacterium liquid 2ml and the 68ml that obtain at every turn and contain 1.2%Al
2O
35% sodium alginate soln mix at normal temperatures, after under aseptic condition, stirring, be added drop-wise to 2% CaCl
2In the aqueous solution, under 4 ℃, left standstill 15 hours.Pachysolen tannophilus ATCC 32691 cells are made gel particles through after the immobilization, and its diameter D is about 3mm.After the immobilization gel particles usefulness sterile water wash three times, change in the proliferated culture medium.
Immobilized pachysolen tannophilus ATCC 32691 is carried out multiplication culture.The proliferated culture medium of 100ml is housed in the triangular flask of 250ml, and the prescription of proliferated culture medium is: wood sugar 20g/L, glucose 50g/L, peptone 3g/L, yeast water 2.5g/L, CaCl
20.25g/L, MgSO
40.25g/L, KH
2PO
42.5g/L.Gel particles is inserted in the 100ml proliferated culture medium, and at 35 ℃, pH5.5 cultivated 36 hours under the condition of 70rpm, changed a fresh medium every 12 hours.
Carry out zymamsis with the pachysolen tannophilus ATCC behind the multiplication culture 32691 at last.The fermention medium of 100ml is housed in the triangular flask of 250ml, and the prescription of fermention medium is: wood sugar 20g/L, glucose 50g/L, CaCl
22.5g/L, MgSO
40.25g/L, KH
2PO
42.5g/L, yeast water 2.5g/L, peptone 3g/L, urea 0.24g/L.Pachysolen tannophilus ATCC behind the multiplication culture 32691 is inserted in the fermention medium of 100ml,, 35 ℃, carried out fermentation reaction under the condition of pH5.5 30 hours at 80rpm.
Measure through high pressure liquid chromatograph: the xylose utilization rate is 76.62%, and the glucose utilization rate has reached 98.08%, and the total reducing sugar utilization ratio is 88.31%.With the ethanol content in the gas chromatographic analysis fermented liquid, the presentation of results alcohol yied has reached 96%.
Embodiment 3
Shehatae candida ATCC 34887 utilizes the method for wood sugar and glucose production alcohol.
The domestication substratum of 100ml is housed in the triangular flask of 250ml, and the prescription of domestication substratum is: wood sugar 50g/L, peptone 6g/L, yeast water 4g/L.With the yeast shehatae candida ATCC34887 that the inclined-plane is preserved, be inoculated in the domestication substratum, inoculum size is 5%.At pH5.5,28 ℃, cultivated under the condition of 120rpm 96 hours, carry out 5 batches domestication altogether and cultivate.With the microscope direct-counting method bacteria containing amount in the nutrient solution is counted, bacteria containing amount increases and obviously increases with domestication batch.The increase of bacteria containing amount illustrates that the ability of its metabolism wood sugar and glucose improves.
Yeast shehatae candida ATCC 34887 after the domestication is carried out liquid culture.The liquid nutrient medium of 100ml is housed in the triangular flask of 250ml, and the prescription of liquid nutrient medium is: wood sugar 50g/L, peptone 6g/L, yeast water 4g/L.Pachysolen tannophilus ATCC 34887 after the domestication is inoculated in the liquid nutrient medium, and inoculum size is 5%.At pH5.5,28 ℃, cultivated 48 hours under the condition of 120rpm, cultivate and finish the back concentration, it is standby to collect nutrient solution.
Use this culture as inoculum then, shehatae candida ATCC 34887 cells are carried out immobilization.Get the step cultivation bacterium liquid 2ml and the 68ml that obtain at every turn and contain 1.2%Al
2O
36% sodium alginate soln mix at normal temperatures, after under aseptic condition, stirring, be added drop-wise to 2% CaCl
2In the aqueous solution, under 4 ℃, left standstill 20 hours.Shehatae candida ATCC 34887 cells are made gel particles through after the immobilization, and its diameter D is about 3mm.After the immobilization gel particles usefulness sterile water wash three times, change in the proliferated culture medium.
Immobilized shehatae candida ATCC 34887 is carried out multiplication culture.The proliferated culture medium of 100ml is housed in the triangular flask of 250ml, and the prescription of proliferated culture medium is: wood sugar 20g/L, glucose 50g/L, peptone 3g/L, yeast water 2.5g/L, CaCl
20.25g/L, MgSO
40.25g/L, KH
2PO
42.5g/L.Gel particles is inserted in the 100ml proliferated culture medium, and at 28 ℃, pH5.5 cultivated 48 hours under the condition of 120rpm, changed a fresh medium every 12 hours.
Carry out zymamsis with the shehatae candida ATCC behind the multiplication culture 34887 at last.The fermention medium of 100ml is housed in the triangular flask of 250ml, and the prescription of fermention medium is: wood sugar 20g/L, glucose 50g/L, CaCl
22.5g/L, MgSO
40.25g/L, KH
2PO
42.5g/L, yeast water 2.5g/L, peptone 3g/L, urea 0.24g/L.Shehatae candida ATCC behind the multiplication culture 34887 is inserted in the fermention medium of 100ml,, 28 ℃, carried out fermentation reaction under the condition of pH value 5.5 36 hours at 120rpm.
Measure through high pressure liquid chromatograph: the xylose utilization rate is 74.68%, and the glucose utilization rate has reached 97.46%, and the total reducing sugar utilization ratio is 87.34%.With the ethanol content in the gas chromatographic analysis fermented liquid, the presentation of results alcohol yied has reached 95%.
Claims (6)
1. a yeast utilizes the method for wood sugar and glucose production alcohol, it is characterized in that this method may further comprise the steps:
1) pachysolen tannophilus 2.1622 be inoculated in the domestication substratum tame;
2) yeast after will taming is inoculated in and carries out liquid culture in the liquid nutrient medium;
3) yeast that liquid culture is obtained carries out immobilization;
4) immobilized yeast is carried out multiplication culture;
5) yeast after the propagation carries out zymamsis;
In the method for above-mentioned production alcohol:
When bacterial strain was tamed, the prescription of domestication substratum was: wood sugar 10-50g/L, and peptone 4-6g/L, yeast water 2-4g/L, the domestication condition is pH5.0-5.5,28-35 ℃, 60-120rpm, each batch domestication time is 48-96 hour.
2. the method for a production alcohol as claimed in claim 1, when it is characterized in that bacterial strain tamed, inoculum size is 2%, domestication is more than 5 batches.
3. the method for a production alcohol as claimed in claim 2, it is characterized in that with the yeast after the domestication prescription of substratum is: wood sugar 10-50g/L, peptone 4-6g/L when carrying out liquid culture, yeast water 2-4g/L, inoculum size is 1-5%, and culture condition is pH5.0-5.5,28-35 ℃, 60-120rpm, incubation time is 24-48 hour, cultivates and finishes the back concentration, and it is standby to collect nutrient solution.
4. the method for a production alcohol as claimed in claim 3, when it is characterized in that the yeast that liquid culture obtains carried out immobilization, at every turn with the concentrated liquid culture of 2ml with contain 1.2%Al
2O
3The 3-6% sodium alginate soln mix at normal temperatures, after under aseptic condition, stirring, be added drop-wise to 2% CaCl
2In the aqueous solution, after leaving standstill 10-20 hour under 4 ℃, make gel particles.
5. the method for a production alcohol as claimed in claim 4, when it is characterized in that fixed yeast cell carried out multiplication culture, the prescription of proliferated culture medium is: wood sugar 20g/L, glucose 50g/L, peptone 3g/L, yeast water 2.5g/L, CaCl
20.25g/L, MgSO
40.25g/L, KH
2PO
42.5g/L the multiplication culture condition is 28-35 ℃, pH5.0-5.5, and 60-120rpm, incubation time were 24-48 hour, changed a fresh medium every 12 hours.
6. the method for a production alcohol as claimed in claim 5, when it is characterized in that yeast behind the multiplication culture carries out zymamsis, the prescription of fermention medium is: wood sugar 20g/L, glucose 50g/L, CaCl
22.5g/L, MgSO
40.25g/L, KH
2PO
42.5g/L, yeast water 2.5g/L, peptone 3g/L, urea 0.24g/L, fermentation condition are pH5.0-5.5,28-35 ℃, 60-120rmp, fermentation time are 24-36 hour.
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| CNB2003101218299A CN100339483C (en) | 2003-12-19 | 2003-12-19 | Process for preparing alcohol through utilizing xylose and glucose by microzyme |
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| CNB2003101218299A CN100339483C (en) | 2003-12-19 | 2003-12-19 | Process for preparing alcohol through utilizing xylose and glucose by microzyme |
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| CN100339483C true CN100339483C (en) | 2007-09-26 |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110056126A1 (en) * | 2009-08-13 | 2011-03-10 | Harvey J T | Process for producing high value products from biomass |
| CN101705253B (en) * | 2009-10-28 | 2012-03-14 | 安徽丰原发酵技术工程研究有限公司 | Method for treating xylose mother solution |
| CN102154128B (en) * | 2010-12-17 | 2012-07-04 | 云南大学 | Pichia anomala strain capable of directly preparing xylose into alcohol |
| CN102174631B (en) * | 2010-12-30 | 2014-01-08 | 吉林省农业科学院 | Method for improving yield of ethanol produced by sugar beet fermentation |
| CN104673712B (en) * | 2015-01-16 | 2021-07-13 | 山东省科学院能源研究所 | Strain for producing alcohol fuel by synchronously utilizing glucose and xylose and application thereof |
| CN114854795B (en) * | 2022-05-11 | 2023-08-18 | 淮阴师范学院 | Method for producing ethanol by double-bacteria fermentation of raw starch |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63219386A (en) * | 1987-03-09 | 1988-09-13 | Hiroaki Horitsu | Production of sugaralcohol |
| CN1141057A (en) * | 1993-11-08 | 1997-01-22 | 普渡研究基金会 | Recombinant yeasts for effective fermentation of glucose and xylose |
| CN1225125A (en) * | 1996-05-06 | 1999-08-04 | 普渡研究基金会 | Stable recombinant yeasts for fermenting xylose to ethanol |
-
2003
- 2003-12-19 CN CNB2003101218299A patent/CN100339483C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63219386A (en) * | 1987-03-09 | 1988-09-13 | Hiroaki Horitsu | Production of sugaralcohol |
| CN1141057A (en) * | 1993-11-08 | 1997-01-22 | 普渡研究基金会 | Recombinant yeasts for effective fermentation of glucose and xylose |
| CN1225125A (en) * | 1996-05-06 | 1999-08-04 | 普渡研究基金会 | Stable recombinant yeasts for fermenting xylose to ethanol |
Non-Patent Citations (3)
| Title |
|---|
| 原料的酒精发酵研究 郑州轻工业学院学报，第18卷第3期 2003 * |
| 发酵木糖酵母菌株的选育 孙玉英，谭海刚等，广州食品工业科技，第18卷第4期 2002 * |
| 木质纤维素酒精发酵菌种的筛选 李素玉，陈新芳，田沈，王菊，杨秀山，太阳能学报，第24卷第2期 2003 * |
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