CN104313106B - Method for preparing ethanol by using binary regulated aureobasidium pullulans and pichia pastoris fermented reed - Google Patents
Method for preparing ethanol by using binary regulated aureobasidium pullulans and pichia pastoris fermented reed Download PDFInfo
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- CN104313106B CN104313106B CN201410536130.7A CN201410536130A CN104313106B CN 104313106 B CN104313106 B CN 104313106B CN 201410536130 A CN201410536130 A CN 201410536130A CN 104313106 B CN104313106 B CN 104313106B
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- aureobasidium pullulans
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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 discloses a method for preparing ethanol by using binary regulated aureobasidium pullulans and pichia pastoris fermented reed and relates to a method for biomass energy ethanol by using reed. The method specifically comprises the following steps: adding 1g/L yeast extract, 0.5g/L MgCl2, 0.5g/L(NH4)2SO4, 0.5g/L KH2PO4, 20g/L glucose and 15g/L xylose into a 50-60g/L dilute sulphuric acid hydrolysate of reed to prepare a culture medium; and by using aureobasidium pullulans and pichia pastoris as fermenting strains, carrying out liquid fermentation by using the culture medium, controlling the pH of initial fermentation at 6.0 and the initial temperature at 25 DEG C, and when fermentation is carried out for 24 hours, respectively regulating the pH and temperature to 4.0 and 30 DEG C, and fermenting to 40 hours to obtain the ethanol. The process for preparing the reed ethanol, which is disclosed by the invention, has the advantages of being environmental friendly and high in fermentation efficiency, and meanwhile, the addition value of reed utilization is further enhanced.
Description
Technical field
The present invention relates to the preparation method and technique of a kind of biomass energy ethanol, belong to biomass energy manufacture field.
Background technology
Phragmites communiss (Phragmites communis) are described as " the second forest " by scientist, in 17 provinces, autonomies of China
Area, municipality directly under the Central Government are distributed, aboundresources, and total amount is produced every year up to more than 200 ten thousand tons, account for the 6% of its Gross World Product.Phragmites communiss
Used as perennial grass family phragmites communiss platymiscium, content of cellulose is up to 40%～60%, is only second to the content of cellulose of linen-cotton, can
Replace timber as paper making raw material, grain can be replaced as the renewable raw materials of biomass energy.
The external exploitation to phragmites communiss it is more be, how by lignin, carbohydrate inversion, beating, preferably to use
In papermaking.It has been to tackle the environment and energy problem for becoming increasingly conspicuous since the eighties in 20 century, European scientific research personnel is attempted phragmites communiss
It is used as a kind of brand-new energy-source plant.German VEBA petrochemical corporation (complex)s plant experimentally 30ha phragmites communiss in 1992, are given birth to it
Hydrogen producing, applies in the polish of crude oil.Szijarto etc. adopts wet oxidation process phragmites communiss, finds 51.7% half fiber
Element and 58.3% lignin are degraded, and cellulose still has 87.1% to exist in solid form, through cellulase water
Xie Hou, it is 82.4% that cellulose is converted into the conversion ratio of glucose, and after synchronized diastatic fermentation, the yield of ethanol is 8.7g/L,
The 73% of theoretical yield is reached, but needs substantial amounts of cellulase preparation assosting effect, production cost is higher.China is at present to reed
The research of reed is concentrated mainly on seeks the sides such as Phragmites communis population biology, ecogenesis process or pulping process and pharmacological action
Face, the development as biomass energy ethanol is at the early-stage.
The content of the invention
In view of this, there is provided a kind of preparation method with inexpensive and efficient reed by ethanol and technique are actually necessary.
A kind of reed by ethanol, is obtained by following methods：
(1) culture medium is configured：Add 1g/L yeast extracts, 0.5g/L in the dilute sulfuric acid hydrolyzed solution of 50-60g/L phragmites communiss
MgCl2、0.5g/L(NH4)2SO4、0.5g/L KH2PO4, 20g/L glucoses, 15g/L xyloses；
(2) fermented bacterium culture：, using Aureobasidium pullulans and Pichia sp., Aureobasidium pullulans are in 28 DEG C of standings for fermented bacterium
Culture 30h, Pichia sp. cultivates 48h in 30 DEG C in the shaking table of 180rpm；
(3) fermentation control：Using Aureobasidium pullulans and Pichia sp. as fermented bacterium, using the culture medium liquid is carried out
Fermentation, pH and temperature, when fermentation is to 24h, are regulated to respectively 4.0 and 30 by control starting fermentation pH6.0,25 DEG C of initial temperature
DEG C, ferment to 40h, ethanol is obtained.
Above-mentioned reed by ethanol is obtained by the dilute acid hydrolysis liquid of Aureobasidium pullulans and Pichia sp. co-fermentation phragmites communiss, Ke Yiwei
The Aureobasidium pullulans and Pichia sp. of free state are inoculated in the culture medium of phragmites communiss hydrolyzed solution to be fermented to 40h and terminate simultaneously；
Can also be that the Aureobasidium pullulans of single free state first ferment phragmites communiss culture medium 24h, the complete red ferment of free state is then accessed again
Mother continues to ferment together to 40h and terminates；Alternatively fixed Aureobasidium pullulans and the Pichia sp. of free state are inoculated in reed simultaneously
Fermented in the culture medium of reed hydrolyzed solution to 40h and terminated；Alternatively fixed Aureobasidium pullulans first ferment phragmites communiss culture medium
24h, the Pichia sp. that free state is then accessed again continues to ferment together to 40h terminations.
The present invention provides a kind of method for preparing reed by ethanol, not only environmentally friendly, with low cost by the technique of its prepared ethanol,
Also improve the added value that phragmites communiss utilize simultaneously.
Specific embodiment
The present invention is further described with reference to specific embodiment
1st, phragmites communiss prepare the embodiment 1 of ethanol
(1) culture medium is configured：Add 1g/L yeast extracts, 0.5g/L in the dilute sulfuric acid hydrolyzed solution of 50g/L phragmites communiss
MgCl2、0.5g/L(NH4)2SO4、0.5g/L KH2PO4, 20g/L glucoses, 15g/L xyloses；
(2) fermented bacterium culture：, using Aureobasidium pullulans and Pichia sp., Aureobasidium pullulans are in 28 DEG C of standings for fermented bacterium
Culture 30h, Pichia sp. cultivates 48h in 30 DEG C in the shaking table of 180rpm；
(3) fermentation control：Using the Aureobasidium pullulans and Pichia sp. of free state as fermented bacterium, while being inoculated in reed
Liquid fermentation is carried out in the culture medium of reed hydrolyzed solution, starting fermentation pH6.0,25 DEG C of initial temperature is controlled, when fermentation is to 24h,
PH and temperature are regulated to into respectively 4.0 and 30 DEG C, are fermented to 40h, 7.7g/L ethanol is obtained.
2nd, phragmites communiss prepare the embodiment 2 of ethanol
(1) culture medium is configured：Add 1g/L yeast extracts, 0.5g/L in the dilute sulfuric acid hydrolyzed solution of 55g/L phragmites communiss
MgCl2、0.5g/L(NH4)2SO4、0.5g/L KH2PO4, 20g/L glucoses, 15g/L xyloses；
(2) fermented bacterium culture：, using Aureobasidium pullulans and Pichia sp., Aureobasidium pullulans are in 28 DEG C of standings for fermented bacterium
Culture 30h, Pichia sp. cultivates 48h in 30 DEG C in the shaking table of 180rpm；
(3) fermentation control：Using the Aureobasidium pullulans and Pichia sp. of free state as fermented bacterium, first individually inoculation trip
In the culture medium of phragmites communiss hydrolyzed solution, control starting fermentation pH6.0,25 DEG C of initial temperature carry out liquid to the Aureobasidium pullulans of amorph
Fermentation, when fermentation is to 24h, by pH and temperature 4.0 and 30 DEG C is regulated to respectively, inoculates the Pichia sp. of free state in fermentation
Ferment together to 40h in liquid, 8.4g/L ethanol is obtained.
3rd, phragmites communiss prepare the embodiment 3 of ethanol
(1) culture medium is configured：Add 1g/L yeast extracts, 0.5g/L in the dilute sulfuric acid hydrolyzed solution of 60g/L phragmites communiss
MgCl2、0.5g/L(NH4)2SO4、0.5g/L KH2PO4, 20g/L glucoses, 15g/L xyloses；
(2) fermented bacterium culture：, using Aureobasidium pullulans and Pichia sp., Aureobasidium pullulans are in 28 DEG C of standings for fermented bacterium
Culture 30h, Pichia sp. cultivates 48h in 30 DEG C in the shaking table of 180rpm；
(3) fermentation control：Using the Pichia sp. of fixed Aureobasidium pullulans and free state as fermented bacterium, while connecing
Plant carries out liquid fermentation in the culture medium of phragmites communiss hydrolyzed solution, control starting fermentation pH6.0,25 DEG C of initial temperature, in fermentation extremely
During 24h, pH and temperature are regulated to into respectively 4.0 and 30 DEG C, are fermented to 40h, 8.9g/L ethanol is obtained.
4th, phragmites communiss prepare the embodiment 4 of ethanol
(1) culture medium is configured：Add 1g/L yeast extracts, 0.5g/L in the dilute sulfuric acid hydrolyzed solution of 60g/L phragmites communiss
MgCl2、0.5g/L(NH4)2SO4、0.5g/L KH2PO4, 20g/L glucoses, 15g/L xyloses；
(2) fermented bacterium culture：, using Aureobasidium pullulans and Pichia sp., Aureobasidium pullulans are in 28 DEG C of standings for fermented bacterium
Culture 30h, Pichia sp. cultivates 48h in 30 DEG C in the shaking table of 180rpm；
(3) fermentation control：Using fixed Aureobasidium pullulans and Pichia sp. as fermented bacterium, first individually inoculation is fixed
Aureobasidium pullulans in the culture medium of phragmites communiss hydrolyzed solution, control starting fermentation pH6.0,25 DEG C of initial temperature carry out liquid and send out
Ferment, when fermentation is to 24h, by pH and temperature 4.0 and 30 DEG C is regulated to respectively, inoculates the Pichia sp. of free state in fermentation liquid
In ferment together to 40h, 9.7g/L ethanol is obtained.
Claims (2)
1. a kind of employing Aureobasidium pullulans joint Pichia sp. two-period form regulating and controlling temperature and pH fermentation phragmites communiss prepare the technique of ethanol,
Including following methods：
(1) culture medium is configured：Add 1g/L yeast extracts, 0.5g/L MgCl in the dilute sulfuric acid hydrolyzed solution of 50-60g/L phragmites communiss2、
0.5g/L(NH4)2SO4、0.5g/L KH2PO4, 20g/L glucoses, 15g/L xyloses；
(2) fermented bacterium culture：, using Aureobasidium pullulans and Pichia sp., Aureobasidium pullulans are in 28 DEG C of quiescent cultures for fermented bacterium
30h, Pichia sp. cultivates 48h in 30 DEG C in the shaking table of 180rpm；
(3) fermentation control：Using Aureobasidium pullulans and Pichia sp. as fermented bacterium, carry out liquid using the culture medium and send out
Ferment, pH and temperature, when fermentation is to 24h, are regulated to respectively 4.0 and 30 by control starting fermentation pH6.0,25 DEG C of initial temperature
DEG C, ferment to 40h, ethanol is obtained.
2. the preparation technology of reed by ethanol as claimed in claim 1, it is characterised in that：It is common by Aureobasidium pullulans and Pichia sp.
The dilute acid hydrolysis liquid of fermentation phragmites communiss, can be that the Aureobasidium pullulans and Pichia sp. of free state are inoculated in phragmites communiss hydrolyzed solution simultaneously
Fermented in culture medium to 40h and terminated；Can also be that the Aureobasidium pullulans of single free state first ferment phragmites communiss culture medium
24h, the Pichia sp. that free state is then accessed again continues to ferment together to 40h terminations；Alternatively fixed Aureobasidium pullulans and
The Pichia sp. of free state is inoculated in the culture medium of phragmites communiss hydrolyzed solution to be fermented to 40h and terminates simultaneously；Alternatively fixed
Aureobasidium pullulans first ferment phragmites communiss culture medium 24h, and the Pichia sp. that free state is then accessed again continues fermentation together to 40h ends
Only.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101497894A (en) * | 2008-01-29 | 2009-08-05 | 中国石油化工集团公司 | Method for preparing ethanol from wood fiber raw material |
| CN102605022A (en) * | 2012-04-13 | 2012-07-25 | 盐城工学院 | Method for co-production of oligose and dietary fiber through wheat bran fermentation |
| CN102876757A (en) * | 2012-10-15 | 2013-01-16 | 南京农业大学 | Technique for preparing (FOs) feruloyl oligosaccharides by adopting two-period-type combined regulation fermentation technology |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101497894A (en) * | 2008-01-29 | 2009-08-05 | 中国石油化工集团公司 | Method for preparing ethanol from wood fiber raw material |
| CN102605022A (en) * | 2012-04-13 | 2012-07-25 | 盐城工学院 | Method for co-production of oligose and dietary fiber through wheat bran fermentation |
| CN102876757A (en) * | 2012-10-15 | 2013-01-16 | 南京农业大学 | Technique for preparing (FOs) feruloyl oligosaccharides by adopting two-period-type combined regulation fermentation technology |
Non-Patent Citations (2)
| Title |
|---|
| 出芽短梗霉发酵合成聚苹果酸的研究;李睿颖等;《食品与发酵科技》;20121031;第47卷(第5期);第2.3节和第3节第1段 * |
| 出芽短梗霉的研究进展;崔堂兵等;《工业微生物》;20020630;第32卷(第2期);第41页第2段 * |
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Effective date of registration: 20191225 Address after: 224300 No. 48, Chaoyang Street, hop Tak Town, Sheyang County, Jiangsu, Yancheng City Patentee after: Sheyang reed development company Address before: 224051 No. 9 Yingbin Road, Jiangsu, Yancheng City Patentee before: Yangcheng Institute of Technology |
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Granted publication date: 20170503 Termination date: 20200926 |