CN1216150C - Method for producing alcohol by solid fermentation of stalks - Google Patents
Method for producing alcohol by solid fermentation of stalks Download PDFInfo
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- CN1216150C CN1216150C CN 02116592 CN02116592A CN1216150C CN 1216150 C CN1216150 C CN 1216150C CN 02116592 CN02116592 CN 02116592 CN 02116592 A CN02116592 A CN 02116592A CN 1216150 C CN1216150 C CN 1216150C
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Abstract
The present invention discloses a synchronous solid state fermentation method for producing alcohol by straws. The present invention has the steps that rolled, cut and pulverized straws carry out steam explosion pretreatment under a constant pressure between 1.2 and 1.4MPa for 5 to 7 min; 42 IU/g of cellulase is added into the pretreated straw powder, 5% of high temperature resistance yeast is vaccinated, and 7.2% of alcohol is obtained through 24 h of fermentation at 45 DEG C; fermentation residue comprises rich protein and partial reduction carbohydrate; after being boiled, the fermentation residue has the fragrance and has the prospects to be used as feedstuff. The present invention has the advantages of low energy consumption, high production efficiency and no environment pollution such as waste slag, waste liquid, etc.
Description
Technical field
This technology relates to a kind of with the method for stalk as raw material production alcohol, particularly a kind of method of producing fuel alcohol with stalk as raw material by solid fermentation.
Background technology
China is agriculture production big country, nearly 600,000,000 tons of the wastes straw of annual all kinds of farm crop, aboundresources, cheap.In theory, cellulosic biomass producing fuel ethyl alcohol by ferment does not have difficulties, but can not carry out suitability for industrialized production for a long time, and this name of the game occurring is that Financial cost is difficult to reach a standard, and does not have high efficiency processing condition.The maximum problem that current China alcohol industry faces is raw material, cost and three problems of pollution.Solve the basic outlet of fermentation alcohol raw material, be to utilize fibrous material to replace grain; Alcohol Production combines with fodder production; Utilize the gene engineering seed selection can direct fermentation the bacterial strain of Mierocrystalline cellulose, hemicellulose, and thermotolerant yeast, anti-assorted bacterium zymamsis bacterium etc.At present, fermentation cellulosic biomass is converted into alcoholic acid research and still has certain gap apart from practical application, causes the reason of this problem that many aspects are arranged, and mainly shows the pretreated energy consumption height of cellulose raw material; The production cost height of cellulase, enzyme is lived low; Cellulase hydrolysis and yeast-leavened optimum temperuture are inharmonious, and zymotechnique is also not overripened; Production concentration is low, causes product recovery cost height.
Summary of the invention
The objective of the invention is to provide a kind of effective method with producing alcohol by solid fermentation of stalks.
Comprise in order to reach the technical scheme that purpose of the present invention takes:
1) pre-treatment of stalk
After rubbing the stalk process also about coarse reduction to 20 order, carry out pre-treatment with steam explosion, the stalk that will handle, pour in the reactor of blasting assembly, feed saturated vapor, boost to 1.2～1.6MPa and pick up counting, react and put down valve in 5～7 minutes, the stalk behind processed in the reactor is recovered in the receptor;
2) preparation of CELLULASE
Select for use koning trichoderma as starting strain, utilize koning trichoderma producing cellulase through solid-state fermentation song, CELLULASE is directly used further in the fermentation synchronously;
(1) koning trichoderma is inoculated into slant medium and cultivates certain hour, scrape a few ring spores from the activatory inclined-plane and insert the sterilized water of being with granulated glass sphere, vibration, dispersal spore is measured spore concentration, makes spore suspension concentration reach 10
6～10
7Individual/mL;
(2) in container, add straw powder and wheat bran, water, ammonium sulfate; Making straw powder and wheat bran weight ratio is 1: 1～1.5: 1, and the solid-liquid weight ratio is 1: 2～1: 3, and the ammonium sulfate addition is 2～2.5% of a gross weight;
(3) with the medium sterilization in (2), the spore suspension of cooling back inoculation gross weight 5～8% obtains CELLULASE after cultivating 72～90 hours under 28 ℃～30 ℃;
3) activated yeast
Select for use can be under 45～50 ℃ of conditions well-grown thermotolerant yeast, the yeast strains one of inoculation wort slant preservation is encircled in the 5mLYEPD liquid nutrient medium, 30 ℃～45 ℃ following 120r/min cultivate 16～20h, and then transfer by 5% inoculum size that shaking table 120r/min cultivates 16～18h in the 250mL triangular flask that 20mLYEPD liquid is housed, enlarge successively;
4) simultaneous saccharification and fermentation
In pretreated straw powder, add CELLULASE, make the cellulase inulinase work in total substrate reach 42～50IU/g, add the water of 2～3 times of weight of solid substrate in the Mierocrystalline cellulose filter paper enzyme activity; Thermotolerant yeast after the inoculation activation, inoculum size is 5～10% of total substrate quality, in 45 ℃～50 ℃ fermentations 20～24 hours, collects alcohol.
Koning trichoderma slant medium (storage medium) used in the above-mentioned processing step can be made like this:
Taking by weighing wheat bran 10g, to add water an amount of, boils 30min, crosses leaching juice, and juice is settled to 100mL, adds agar powder 2g, and packing sterilization, cooling tilt to place.
The present invention has realized that from the angle of stalk synchronous glycosylation fermentation the solid-state synchronous fermentation of stalk is converted into the process of alcohol, has following characteristics and advantage:
The present invention selects resistant to elevated temperatures yeast for use, has solved the inconsistent technical barrier of optimum temperuture of yeast optimum growth temperature and cellulase hydrolysis, has satisfied the requirement of synchronous fermentation;
Utilize koning trichoderma producing cellulase through solid-state fermentation song, CELLULASE is directly used further in the fermentation synchronously, not only can realize the synchronous producing alcohol by solid-state fermentation of stalk, and saved the process of purifying, preparation cellulase preparation, the substratum that while cellulase-producing song is utilized is based on stalk, only add on a small quantity wheat bran and ammonium sulfate cheaply, not only utilized stalk, and greatly reduce the cost of preparation enzyme;
By research to synchronous solid state fermentation conditions, obtained the zymotechnique route of optimizing, shortened fermentation time greatly, improved fermentation efficiency, fermented synchronously 20～24 hours, can obtain the alcohol (pressing the stalk butt calculates) about 7.2%;
Utilize this technology not only agricultural wastes such as stalk can be converted into fuel alcohol, and whole technological process energy consumption is low, contaminate environment such as no waste residue, waste liquid are the utilization of waste material methods of environmental protection;
Contain abundant cellulose substances in the stalk, produce alcohol with stalk as fermenting raw materials, not only can fundamentally change present alcohol is the production status of raw material with the starchy material, and can obtain the lees feeds of super quality and competitive price simultaneously, and great economic benefit and social benefit are arranged.The fermentation residue that utilizes this technology to produce contains more rich protein and partial reduction carbohydrate, and smell delicate fragrance after the boiling has the application prospect as feed.
Description of drawings
Fig. 1 is the cultural method process flow sheet of CELLULASE among the present invention;
Fig. 2 is the process flow sheet of the present invention with the synchronous producing alcohol by solid-state fermentation method of stalk.
Specific embodiments
Scheme one: as shown in Figure 1 and Figure 2,
(1) stalk is cut pulverizer and rubbed and coarse reduction to 20 order with rubbing; With the stalk after pulverizing, pour in the reactor of blasting assembly, feed saturated vapor, boost to 1.2MPa and pick up counting, react and put down valve in 5 minutes, stalk is recovered in the receptor;
(2) koning trichoderma being inoculated into slant medium, to be cultured to mycelial growth vigorous, scrapes the sterilized water that a few ring spores insert the band granulated glass spherees vibration, dispersal spore from the inclined-plane; Spore suspension concentration is 10
7Individual/mL;
(3) in container, add straw powder and wheat bran, water, ammonium sulfate; Making straw powder and wheat bran weight ratio is 1: 1, and the solid-liquid weight ratio is 1: 3, and the ammonium sulfate addition is 2% of a gross weight, mix, and sterilization, the spore suspension of cooling back inoculation gross weight 5% obtains CELLULASE after cultivating 84 hours under 30 ℃;
(4) the thermotolerant yeast strain one of inoculation wort slant preservation encircles in the 5mLYEPD liquid nutrient medium, and 45 ℃ of following 120r/min cultivate 16h, and then enlarge successively by 5% inoculum size;
(5) add CELLULASE in the straw powder after explosion, make the cellulase inulinase work in total substrate reach 42IU/g in the Mierocrystalline cellulose filter paper enzyme activity, the water that adds 2 times of weight of solid substrate, thermotolerant yeast after the inoculation activation, inoculum size is 5% of total substrate quality, in 45 ℃ of fermentations 24 hours, collect alcohol, can obtain every 100g dry straw and produce alcohol 7.23g approximately.
Scheme two:
(1) stalk is rubbed also coarse reduction; With the stalk after pulverizing, pour in the reactor of blasting assembly, feed saturated vapor, boost to 1.2MPa and pick up counting, react and put down valve in 5 minutes, stalk is recovered in the receptor;
(2) scrape a few ring spores from the koning trichoderma slant medium and insert the sterilized water, vibration, dispersal spore, spore suspension concentration is 10
6Individual/mL;
(3) in container, add straw powder and wheat bran, water, ammonium sulfate; Making straw powder and wheat bran weight ratio is 1: 1, and the solid-liquid weight ratio is 1: 2, and the ammonium sulfate addition is 2.5% of a gross weight, mix, and sterilization, the spore suspension of cooling back inoculation gross weight 5% obtains CELLULASE after cultivating 76 hours under 30 ℃;
(4) the thermotolerant yeast strain one of inoculation wort slant preservation encircles in the 5mLYEPD liquid nutrient medium, and 30 ℃ of following 120r/min cultivate 18h, and then enlarge successively by 5% inoculum size;
(5) add CELLULASE in the straw powder after explosion, make the cellulase inulinase work in total substrate reach 47IU/g, add the water of 2 times of weight of solid substrate in the Mierocrystalline cellulose filter paper enzyme activity; Thermotolerant yeast after the inoculation activation, inoculum size is 5% of total substrate quality, in 50 ℃ of fermentations 24 hours, collects alcohol, can obtain every 100g dry straw and produce alcohol 7.14g approximately.
Claims (4)
1, a kind of method with producing alcohol by solid fermentation of stalks comprises following production stage:
1) pre-treatment of stalk
With stalk through rubbing and coarse reduction after, carry out pre-treatment with steam explosion, the stalk that will handle, pour in the reactor of blasting assembly, feed saturated vapor, boost to 1.2～1.6MPa and pick up counting, react and put down valve in 5～7 minutes, the stalk behind processed in the reactor is recovered in the receptor;
2) preparation of CELLULASE
Select for use koning trichoderma as starting strain, utilize koning trichoderma producing cellulase through solid-state fermentation song, CELLULASE is directly used further in the fermentation synchronously;
(1) koning trichoderma is inoculated into slant medium and cultivates certain hour, scrape a few ring spores from the activatory inclined-plane and insert the sterilized water of being with granulated glass sphere, vibration, dispersal spore is measured spore concentration, makes spore suspension concentration reach 10
6～10
7Individual/mL;
(2) in container, add straw powder and wheat bran, water and ammonium sulfate; Making straw powder and wheat bran weight ratio is 1: 1～1.5: 1, and the solid-liquid weight ratio is 1: 2～1: 3, and the ammonium sulfate addition is 2～2.5% of a gross weight;
(3) with the medium sterilization in (2), the spore suspension of cooling back inoculation gross weight 5～8% obtains CELLULASE after cultivating 72～90 hours under 28 ℃～30 ℃;
3) activated yeast
Select for use can be under 45～50 ℃ of conditions well-grown thermotolerant yeast, the yeast strains one of inoculation wort slant preservation is encircled in the 5mLYEPD liquid nutrient medium, 30 ℃～45 ℃ following 120r/min cultivate 16～20h, and then transfer by 5% inoculum size that shaking table 120r/min cultivates 16～18h in the 250mL triangular flask that 20mLYEPD liquid is housed, enlarge successively;
4) simultaneous saccharification and fermentation
In pretreated straw powder, add CELLULASE, make the cellulase inulinase work in total substrate reach 42～50IU/g, add the water of 2～3 times of weight of solid substrate in the Mierocrystalline cellulose filter paper enzyme activity; Thermotolerant yeast after the inoculation activation, inoculum size is 5～10% of total substrate quality, in 45 ℃～50 ℃ fermentations 20～24 hours, collects alcohol.
2, the method with producing alcohol by solid fermentation of stalks according to claim 1 is characterized in that koning trichoderma slant medium used in this method is made like this:
Taking by weighing wheat bran 10g, to add water an amount of, boils 30min, crosses leaching juice, and juice is settled to 100mL, adds agar powder 2g, and packing sterilization, cooling tilt to place.
3, the method with producing alcohol by solid fermentation of stalks according to claim 1 is characterized in that
(1) stalk is cut pulverizer and rubbed and coarse reduction to 20 order with rubbing; With the stalk after pulverizing, pour in the reactor of blasting assembly, feed saturated vapor, boost to 1.2MPa and pick up counting, react and put down valve in 5 minutes, stalk is recovered in the receptor;
(2) koning trichoderma being inoculated into slant medium, to be cultured to mycelial growth vigorous, scrapes the sterilized water that a few ring spores insert the band granulated glass spherees vibration, dispersal spore from the inclined-plane; Spore suspension concentration is 10
7Individual/mL;
(3) in container, add straw powder and wheat bran, water, ammonium sulfate; Making straw powder and wheat bran weight ratio is 1: 1, and the solid-liquid weight ratio is 1: 3, and the ammonium sulfate addition is 2% of a gross weight, mix, and sterilization, the spore suspension of cooling back inoculation gross weight 5% obtains CELLULASE after cultivating 84 hours under 30 ℃;
(4) the thermotolerant yeast strain one of inoculation wort slant preservation encircles in the 5mLYEPD liquid nutrient medium, and 45 ℃ of following 120r/min cultivate 16h, and then enlarge successively by 5% inoculum size;
(5) add CELLULASE in the straw powder after explosion, make the cellulase inulinase work in total substrate reach 42IU/g in the Mierocrystalline cellulose filter paper enzyme activity, the water that adds 2 times of weight of solid substrate, thermotolerant yeast after the inoculation activation, inoculum size is 5% of total substrate quality, in 45 ℃ of fermentations 24 hours, collect alcohol, can obtain every 100g dry straw and produce alcohol 7.23g approximately.
4, the method with producing alcohol by solid fermentation of stalks according to claim 1 is characterized in that,
(1) stalk is rubbed also coarse reduction; With the stalk after pulverizing, pour in the reactor of blasting assembly, feed saturated vapor, boost to 1.2MPa and pick up counting, react and put down valve in 5 minutes, stalk is recovered in the receptor;
(2) scrape a few ring spores from the koning trichoderma slant medium and insert the sterilized water, vibration, dispersal spore, spore suspension concentration is 10
6Individual/mL;
(3) in container, add straw powder and wheat bran, water, ammonium sulfate; Making straw powder and wheat bran weight ratio is 1: 1, and the solid-liquid weight ratio is 1: 2, and the ammonium sulfate addition is 2.5% of a gross weight, mix, and sterilization, the spore suspension of cooling back inoculation gross weight 5% obtains CELLULASE after cultivating 76 hours under 30 ℃;
(4) the thermotolerant yeast strain one of inoculation wort slant preservation encircles in the 5mLYEPD liquid nutrient medium, and 30 ℃ of following 120r/min cultivate 18h, and then enlarge successively by 5% inoculum size;
(5) add CELLULASE in the straw powder after explosion, make the cellulase inulinase work in total substrate reach 47IU/g, add the water of 2 times of weight of solid substrate in the Mierocrystalline cellulose filter paper enzyme activity; Thermotolerant yeast after the inoculation activation, inoculum size is 5% of total substrate quality, in 50 ℃ of fermentations 24 hours, collects alcohol, can obtain every 100g dry straw and produce alcohol 7.14g approximately.
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| CN 02116592 CN1216150C (en) | 2002-04-12 | 2002-04-12 | Method for producing alcohol by solid fermentation of stalks |
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|---|---|---|---|
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2002
- 2002-04-12 CN CN 02116592 patent/CN1216150C/en not_active Expired - Fee Related
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