CN107177637A - The methods and applications pre-processed using microorganism to maize straw hydrolysis and saccharification - Google Patents
The methods and applications pre-processed using microorganism to maize straw hydrolysis and saccharification Download PDFInfo
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- CN107177637A CN107177637A CN201710453769.2A CN201710453769A CN107177637A CN 107177637 A CN107177637 A CN 107177637A CN 201710453769 A CN201710453769 A CN 201710453769A CN 107177637 A CN107177637 A CN 107177637A
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- maize straw
- saccharification
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
- C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2201/00—Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
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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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- Chemical Kinetics & Catalysis (AREA)
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- General Chemical & Material Sciences (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention relates to lignocellulose raw material production ethanol field, the methods and applications that especially a kind of utilization microorganism pre-processes to maize straw hydrolysis and saccharification.This method comprises the following steps：(1) by after corn straw smashing, sieving is placed in standby in drier；(2) maize straw is put in triangular flask, after high pressure steam sterilization, access Coriolus Versicolor and oyster cap fungus mixed bacteria liquid, sterile inorganic salts and land plaster, it is eventually adding sterilized water, it is put into after mixing in constant incubator and carries out microculture, after culture is finished, maize straw is filtered by vacuum, is cleaned and dried successively, the maize straw pre-processed.Using pretreated stalk reduced sugar output increased 32% of the invention, its method is gentle to condition requirement, low for equipment requirements without high-temperature high-pressure apparatus, low energy consumption and does not produce poisonous and harmful, is adapted to industrialized production.
Description
Technical field
It is especially a kind of to utilize microorganism to maize straw the present invention relates to lignocellulose raw material production ethanol field
The methods and applications of hydrolysis and saccharification pretreatment.
Background technology
The main substitute that alcohol has become the conventional fossil fuel energy is produced using stalk, but due to wooden in stalk
Biopolymer is interweaved the design feature being closely connected in cellulose, and it is difficult to improve to make its efficiency hydrolyzed.In recent years, people
In the production process in edible mushroom, it is found that oyster cap fungus can to a certain extent can be with the lignin in degrading straw.
In nature, Coriolus Versicolor is the higher bacterial strain of degrading straw efficiency.
At present, ethanol has received much concern as safety, clean fuel and gasoline additive, greatly develops alternative biography
The New-type fuel of system gasoline is imperative.And ethanol petrol is exactly a good solution, traditional alcohol fermentation industry
Mainly using the foodstuff starch such as corn, wheat or sugar-cane juice as raw material, but its cost of material is up to the 40% of totle drilling cost.And China ground
National conditions present situation more than few people, also determines to cause " grain to be striven with people and animals, with grain for the ethanol large-scale production of raw material based on grain
Food strive ground " unfavorable situation.Also will be by land resource and raw material using the sugar crops such as sugarcane production alcohol fuel in China
The limitation of cost and can not large-scale promotion.Biomass is a kind of important renewable resource, and fuel second is produced using biomass
Alcohol is a kind of new energy utilization type, and environmental pollution is small, is current study hotspot.China's biomass material enriches, wherein
Maize straw annual production is up to more than 700,000,000 tons, accounts for the 35% of biomass material.If biomass resource can be converted into available
Liquid or gaseous fuel, by it is strong alleviate crisis of resource, energy shortage and the environmental pollution that the mankind are faced a series of problems, such as,
Possesses good application prospect.
One of main method by fuel ethanol produced by straw is then to utilize to make wine micro- by microbial hydrolytic production sugar
Biological production alcohol fuel.But because the be interweaved structure that is closely connected of biopolymer is special in lignocellulosic in stalk
Point, it is difficult to improve to make its efficiency hydrolyzed.Coniferyl alcohol, sinapinic alcohol are typically contained in lignin and to three kinds of different types of tonquinol
Phenylpropane monomers, they with plant variety, plant age even cell membrane form position difference and exist different
Ratio.Because the copolymerization of this three classes alcohol produces uneven, without optical activity, intersection bonding, high dispersion and high resistance
The aromatic polymer of biological degradability, the attack to most microorganisms is resistant.Therefore, the decomposition of lignin is special
Slowly, and in complete lignum, lignin is present with a kind of physical barriers, and it surrounds cellulose, as periphery
Matrix, protection cellulose exempts from microorganism attack and the attack of digestive enzyme, and its degraded must be prior to cellulose decomposition.
The content of the invention
A kind of pre-processed to maize straw hydrolysis and saccharification it is an object of the invention to provide utilization microorganism method and should
With, by using microorganism to stalk carry out pretreatment production alcohol, for stalk production alcohol new manufacturing technique method is provided.
In order to achieve the above object, the technical scheme is that：
A kind of method that utilization microorganism pre-processes to maize straw hydrolysis and saccharification, comprises the following steps：
(1) by after corn straw smashing, sieving is placed in standby in drier；
(2) maize straw is put in triangular flask, after autoclaving, access Coriolus Versicolor and oyster cap fungus mixed bacteria liquid, nothing
Bacterium inorganic salts and land plaster, are eventually adding to be put into constant incubator after sterilized water, mixing and carry out microculture, wait to cultivate
After finishing, maize straw is filtered by vacuum, is cleaned and dried successively, the maize straw pre-processed.
In the method that described utilization microorganism pre-processes to maize straw hydrolysis and saccharification, step (2), reaction system is consolidated
Liquid mass ratio is 1：5~50.
In the method that described utilization microorganism pre-processes to maize straw hydrolysis and saccharification, step (2), reaction system it is mixed
It is 10mL~100mL/L to close bacterium solution inoculum concentration.
It is miscellaneous in mixed bacteria liquid in the method that described utilization microorganism pre-processes to maize straw hydrolysis and saccharification, step (2)
Color rainbow conk and the mixing quality ratio of oyster cap fungus are 1:0.1~10.
In the method that described utilization microorganism pre-processes to maize straw hydrolysis and saccharification, step (2), the species of inorganic salts
For KH2PO4、MgSO4、FeSO4、CaCl2One or more, the mass ratio of stalk and inorganic salts is 10~150：1, straw
The mass ratio of stalk and land plaster is 50~200：1.
In the method that described utilization microorganism pre-processes to maize straw hydrolysis and saccharification, step (2), microculture
Temperature is 20 DEG C~50 DEG C, and the time of microculture is 7 days~30 days.
The application that described utilization microorganism pre-processes to maize straw hydrolysis and saccharification, the maize straw of pretreatment is in biology
Applied in matter alcohol fuel or biomass ferment engineering.
The present invention design philosophy be：
In recent years, people are in the production process in edible mushroom, it is found that oyster cap fungus can degrade straw to a certain extent
Lignin in stalk.In nature, Coriolus Versicolor is the higher bacterial strain of degrading straw efficiency.The present invention for oyster cap fungus and
The characteristic of Coriolus Versicolor, according to the two influence under mixing condition to stalk hydrolysis and saccharification processing procedure, probes into out corn stalk
The optimum process condition of stalk hydrolysis and saccharification in the case where double bacterium act on, it is intended to provide new manufacturing technique method for stalk production alcohol.
Being mainly by cellulose, hemicellulose with lignin by covalent bonding junction into network structure for stalk raw materials, causes fiber
Plain follow-up saccharification efficiency is low, and alcohol yied is low.It is therefore desirable to which by pretreatment, destruction lignin is fine to cellulose and half
The parcel of element is tieed up, the crystallinity of cellulose is reduced, follow-up saccharification efficiency is improved, and then improve biomass economy rate.
Advantages of the present invention and beneficial effect are：
(1) the inventive method is gentle to condition requirement, low for equipment requirements without high-temperature high-pressure apparatus, low energy consumption and not
Produce poisonous and harmful, suitable industrialized production.
(2) low production cost of the present invention, can be used for a long time after culture presevation.
(3) present invention does not produce poisonous and harmful waste, smaller to environmental hazard.
(4) pretreated stalk reduced sugar output increased 32% of the invention is utilized, 37% is can reach.
Brief description of the drawings
Fig. 1 is the schematic flow sheet of lignin degradation of the present invention.
Embodiment
In specific implementation process, the method pre-processed using microorganism to maize straw hydrolysis and saccharification, including following step
Suddenly：
(1) maize straw is chosen, sieving after crushed is chosen；
(2) maize straw is placed in sterilization treatment in triangular flask, then accesses Coriolus Versicolor and oyster cap fungus, 24 DEG C of bars
Cultivated 7~20 days under part, after culture terminates, maize straw is filtered by vacuum, is cleaned and dried successively, is pre-processed
Maize straw, then through hydrolysis production sugar.
The carbon containing, methoxyl group of Coriolus Versicolor lignin degrading product, hydrogen are fewer than substrate element, and contain compared with polyoxy, carbonyl, relatively
For armaticity reduce, α β unsaturated acyclic hydrocarbons increase, but approached with the molecular weight of former substrate.Therefore, according to its catabolite point
Analysis, the process of lignin degradation can be divided into：Demethylation and hydroxyl simultaneously form polyphenol class formation；Oxygenation cracks polyphenol ring, produces chain
Hydrocarbon；Hydrolysis shortens aliphatic hydrocarbon.Enzyme system is that extracellular or be bound by cell membrane, its enzyme system produced mainly includes laccase
(Lac), peroxidase (LiP) and manganese compound enzyme (MnP).
As shown in figure 1, the preprocess method of maize straw production ethanol of the present invention, maize straw and Fenton reagent is main
Course of reaction is as follows：
Peroxidase+H2O2+ the H of → multienzyme complex I2O (1)
+ the SH of multienzyme complex I2→ SH*+ multienzyme complexs II (2)
+ the SH of multienzyme complex II2→ SH*+ peroxidase (3)
2SH*+2 (lignin) → 2 (lignin free radical)+2SH2 (4)
Overall reaction：H2O2+ 2 (lignin) → 2 (lignin free radical)+H2O
The present invention is expanded on further with reference to embodiment, it should be appreciated that embodiment described herein
It is merely to illustrate and explain the present invention, is not intended to limit the invention.
Embodiment 1
In the present embodiment, the method pre-processed using microorganism to maize straw hydrolysis and saccharification is comprised the following steps：
1st, by after corn straw smashing, 40 mesh sieves is crossed, are placed in standby in drier.
2nd, 10g stalks are weighed to be placed in triangular flask, are gone out after being sealed with gauze and brown paper with 121 DEG C of high pressure 100kPa steam
Bacterium 30 minutes.
3rd, 10mL Coriolus Versicolors and oyster cap fungus mixed bacteria liquid (the mixing matter of Coriolus Versicolor and oyster cap fungus are added into bottle
Amount ratio is 1:1), 0.3 gram of KH2PO4, 0.1 gram of land plaster, and add sterilized water and mend to 100mL.Reacted 7 days under the conditions of 24 DEG C,
105 DEG C of drying, the maize straw pre-processed after vacuum filtration, cleaning.
Embodiment 2
In the present embodiment, the method pre-processed using microorganism to maize straw hydrolysis and saccharification is comprised the following steps：
1st, by after corn straw smashing, 40 mesh sieves is crossed, are placed in standby in drier.
2nd, 10g stalks are weighed to be placed in triangular flask, are gone out after being sealed with gauze and brown paper with 121 DEG C of high pressure 100kPa steam
Bacterium 30 minutes.
3rd, 20mL Coriolus Versicolors and oyster cap fungus mixed bacteria liquid (the mixing matter of Coriolus Versicolor and oyster cap fungus are added into bottle
Amount ratio is 1:2), 0.3 gram of MgSO4, 0.1 gram of land plaster, and add sterilized water and mend to 100mL.Reacted 7 days under the conditions of 24 DEG C,
105 DEG C of drying, the maize straw pre-processed after vacuum filtration, cleaning.
Embodiment 3
In the present embodiment, the method pre-processed using microorganism to maize straw hydrolysis and saccharification is comprised the following steps：
1st, by after corn straw smashing, 40 mesh sieves is crossed, are placed in standby in drier.
2nd, 10g stalks are weighed to be placed in triangular flask, are gone out after being sealed with gauze and brown paper with 121 DEG C of high pressure 100kPa steam
Bacterium 30 minutes.
3rd, 30mL Coriolus Versicolors and oyster cap fungus mixed bacteria liquid (the mixing matter of Coriolus Versicolor and oyster cap fungus are added into bottle
Amount ratio is 1:3), 0.3 gram of CaCl2, 0.1 gram of land plaster, and add sterilized water and mend to 100mL.Reacted 7 days under the conditions of 24 DEG C,
105 DEG C of drying, the maize straw pre-processed after vacuum filtration, cleaning.
Embodiment 4
In the present embodiment, the method pre-processed using microorganism to maize straw hydrolysis and saccharification is comprised the following steps：
1st, by after corn straw smashing, 40 mesh sieves is crossed, are placed in standby in drier.
2nd, 10g stalks are weighed to be placed in triangular flask, are gone out after being sealed with gauze and brown paper with 121 DEG C of high pressure 100kPa steam
Bacterium 30 minutes.
3rd, 40mL Coriolus Versicolors and oyster cap fungus mixed bacteria liquid (the mixing matter of Coriolus Versicolor and oyster cap fungus are added into bottle
Amount ratio is 1:0.6), 0.3 gram of FeSO4, 0.1 gram of land plaster, and add sterilized water and mend to 100mL.7 are reacted under the conditions of 24 DEG C
My god, 105 DEG C of drying, the maize straw pre-processed after vacuum filtration, cleaning.
Embodiment 5
In the present embodiment, the method pre-processed using microorganism to maize straw hydrolysis and saccharification is comprised the following steps：
1st, by after corn straw smashing, 40 mesh sieves is crossed, are placed in standby in drier.
2nd, 10g stalks are weighed to be placed in triangular flask, are gone out after being sealed with gauze and brown paper with 121 DEG C of high pressure 100kPa steam
Bacterium 30 minutes.
3rd, 50mL Coriolus Versicolors and oyster cap fungus mixed bacteria liquid (the mixing matter of Coriolus Versicolor and oyster cap fungus are added into bottle
Amount ratio is 1:0.3), 0.2 gram of CaCl2, 0.1 gram of MgSO4, 0.1 gram of land plaster, and add sterilized water and mend to 100mL.24 DEG C of bars
Reacted 7 days under part, 105 DEG C of drying, the maize straw pre-processed after vacuum filtration, cleaning.
Application examples
The maize straw pre-processed in embodiment 1 to 5 is applied in biomass fuel ethanol or biomass ferment engineering
In, experimental data and technique effect are as follows：
The yield for mixing pretreatment maize straw production reduced sugar with the double bacterium of Coriolus Versicolor and oyster cap fungus is tested, in production sugar
When under the microbial inoculum addition 30mL/L treatment conditions on peak, of a relatively high Reducing sugar is 20.1%, and reduction candy output can be carried
It is high by 32%.The mixed bacteria liquid pretreated straw reduced sugar candy output of different adding proportions is shown in Table 1.
Reduced sugar candy output changes before and after the different mixed bacteria liquid addition pretreated straws of table 1
Embodiment and application examples result show, by the preprocess method of maize straw of the present invention, Coriolus Versicolor with it is rough
Under the collective effect that skin is picked up the ears, lignin is substantially destroyed to the parcel of cellulose hemicellulose in stalk, so that in certain journey
Improved on degree production and alcohol saccharification pretreatment stage sugared rate, this is to produce alcohol using stalk to provide new technique on the way
Footpath and processing method.Also, stalk can be more utilized in edible mushroom actual production, and will be given up after Edible Fungi
The oyster cap fungus section abandoned reclaims the raw material as production alcohol.Working condition of the present invention is gentle, with low cost, can after culture presevation
Use for a long time, and do not produce poisonous and harmful waste, it is smaller to environmental hazard, resource utilization is not only improved, can also be
Alleviate the pressure that energy scarcity is brought to a certain extent.
Claims (7)
1. a kind of method that utilization microorganism pre-processes to maize straw hydrolysis and saccharification, it is characterised in that comprise the following steps：
(1) by after corn straw smashing, sieving is placed in standby in drier；
(2) maize straw is put in triangular flask, after autoclaving, access Coriolus Versicolor and oyster cap fungus mixed bacteria liquid, sterile nothing
Machine salt and land plaster, are eventually adding to be put into constant incubator after sterilized water, mixing and carry out microculture, treat that culture is finished
Afterwards, maize straw is filtered by vacuum, is cleaned and dried successively, the maize straw pre-processed.
2. the method that utilization microorganism according to claim 1 pre-processes to maize straw hydrolysis and saccharification, it is characterised in that：
In step (2), the solid-liquid mass ratio example of reaction system is 1：5~50.
3. the method that utilization microorganism according to claim 1 pre-processes to maize straw hydrolysis and saccharification, it is characterised in that：
In step (2), the mixed bacteria liquid inoculum concentration of reaction system is 10mL~100mL/L.
4. the method that utilization microorganism according to claim 1 pre-processes to maize straw hydrolysis and saccharification, it is characterised in that：
In step (2), the mixing quality ratio of Coriolus Versicolor and oyster cap fungus is 1 in mixed bacteria liquid:0.1~10.
5. the method that utilization microorganism according to claim 1 pre-processes to maize straw hydrolysis and saccharification, it is characterised in that：
In step (2), the species of inorganic salts is KH2PO4、MgSO4、FeSO4、CaCl2One or more, stalk and inorganic salts
Mass ratio is 10~150：1, the mass ratio of stalk and land plaster is 50~200：1.
6. the method that utilization microorganism according to claim 1 pre-processes to maize straw hydrolysis and saccharification, it is characterised in that：
In step (2), the temperature of microculture is 20 DEG C~50 DEG C, and the time of microculture is 7 days~30 days.
7. the application that the utilization microorganism described in a kind of one of claim 1 to 6 pre-processes to maize straw hydrolysis and saccharification, it is special
Levy and be：The maize straw of pretreatment is applied in biomass fuel ethanol or biomass ferment engineering.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108728500A (en) * | 2018-06-26 | 2018-11-02 | 深圳崀途科技有限公司 | A kind of technique that alcohol fuel is prepared using stalk |
| CN112226473A (en) * | 2020-07-15 | 2021-01-15 | 湖北大学 | Method for improving enzymolysis efficiency of corn straw by using rhodococcus bifidus |
| CN114908133A (en) * | 2022-04-19 | 2022-08-16 | 天津商业大学 | Method for improving yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment |
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| CN1880415A (en) * | 2005-06-14 | 2006-12-20 | 河南农业大学 | Process for preparing fuel ethanol by using straw fiber materials |
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| CN1880415A (en) * | 2005-06-14 | 2006-12-20 | 河南农业大学 | Process for preparing fuel ethanol by using straw fiber materials |
| CN102870598A (en) * | 2012-09-28 | 2013-01-16 | 安徽农业大学 | Method for screening Pleurotus ostreatus strains capable of efficiently degrading cotton stalk lignin |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108728500A (en) * | 2018-06-26 | 2018-11-02 | 深圳崀途科技有限公司 | A kind of technique that alcohol fuel is prepared using stalk |
| CN112226473A (en) * | 2020-07-15 | 2021-01-15 | 湖北大学 | Method for improving enzymolysis efficiency of corn straw by using rhodococcus bifidus |
| CN114908133A (en) * | 2022-04-19 | 2022-08-16 | 天津商业大学 | Method for improving yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment |
| CN114908133B (en) * | 2022-04-19 | 2024-02-13 | 天津商业大学 | Method for improving reducing sugar produced by corn straw hydrolysis through biological combined ammoniation pretreatment |
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Application publication date: 20170919 |