CN107916279A - A kind of detoxification fermentation method in situ of ligno-cellulose hydrolysate - Google Patents
A kind of detoxification fermentation method in situ of ligno-cellulose hydrolysate Download PDFInfo
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- CN107916279A CN107916279A CN201610886906.7A CN201610886906A CN107916279A CN 107916279 A CN107916279 A CN 107916279A CN 201610886906 A CN201610886906 A CN 201610886906A CN 107916279 A CN107916279 A CN 107916279A
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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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- 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 present invention provides a kind of detoxification fermentation process in situ of ligno-cellulose hydrolysate.This method is：Appropriate lignin is added in ligno-cellulose hydrolysate as detoxification reagent, carries out detoxification in situ to zymotic fluid during the fermentation, suppression of the toxic chemical present in ligno-cellulose hydrolysate to alcohol fermentation is being reduced, is improving alcohol production efficiency.The present invention is without carrying out extra detoxification treatment to lignocellulosic Soviet Union hydrolyzate, saving equipment investment；Detoxification reagent source is extensive, of low cost, recoverable.The invention has the advantages that：(1) technique is simple and convenient to operate.(2) operating time is short, equipment investment is low, saves the energy；(3) effectively improve alcohol fermentation efficiency, reduce industrial production cost, be of great significance to the cost for reducing Production of Alcohol from Lignocellulose.
Description
Technical field
Alcohol fuel field is produced by zymotechnique the present invention relates to biomass material degraded, and in particular to a kind of wooden
The detoxification fermentation method in situ of cellulosic hydrolysate.
Background technology
With the increasingly depleted of fossil energy and increasingly sharpening for environmental pollution, renewable and clean energy resource alcohol fuel is opened
Send out and utilize the extensive concern for receiving people.For traditional alcohol fermentation using sugar or starch as raw material, the two is all the master of food
Source is wanted, producing alcohol fuel as raw material using grain constitutes safely world food threat, finds other raw materials and substitutes
Grain is imperative.The lignocellulosic renewable resource the abundantest and cheap as nature, its main component cellulose
Hemicellulose is the raw materials for production of potential alcohol fuel, is studied using lignocellulosic materials for fuel ethanol as countries in the world
Hot spot.
But cellulose, hemicellulose and lignin three link are close in ligno-cellulosic materials, only to wooden fibre
The plain raw material of dimension is pre-processed, the structure of the lignin of destruction height polymerization, cellulose and hemicellulose, its is carried out enzyme
Hydrolysis obtains fermentable sugars.The conditions such as the high temperature of the heat-chemistry preprocessing process such as acidolysis or steam blasting, high pressure, low pH can make
Generated in hydrolyzate or discharge the inhibiting factors such as organic acid, furans and phenolic compound, these inhibiting factors are to ferment of making wine
Mother's fermentation produces strong inhibition effect, so as to influence the normal growth of saccharomycete and subsequent fermentation process.
The method of traditional removal inhibiting factor includes activated carbon adsorption, ion exchange, calcium hydroxide neutralization, peroxidase
Processing etc..But often cost is higher for these methods, and substantial amounts of waste water or solid waste are produced in processing procedure.Also very
More research reports reduce the mortifier concentration in hydrolyzate, or improvement hair by optimizing lignin matter preprocessing process
Yeast-like fungi kind can all increase considerably cost and still be in development phase to the tolerance of mortifier, these methods.
The content of the invention
For above-mentioned technical problem, the present invention provides a kind of detoxification in situ fermentation of lignocellulose biomass hydrolyzate
New method, can effectively reduce influence of the mortifier to fermentation, improve lignocellulose biomass producing fuel ethyl alcohol by ferment
Efficiency.
A kind of detoxification fermentation process in situ of lignocellulose biomass hydrolyzate, the technical solution of use are：
(1) the hydrolysis slurries that the lignocellulosic of pretreatment obtains after degraded；
(2) the hydrolysis slurries of acquisition are subjected to filtering and remove solid residue, obtain ligno-cellulose hydrolysate；
(3) and then into ligno-cellulose hydrolysate lignin, saccharomyces cerevisiae progress fermentation process are added, obtains fuel second
Alcohol.
The preprocess method that the lignocellulosic of pretreatment described in step (1) is applicable in includes chemical method, Physical, change
- Physical and bioanalysis etc..
The biodegrading process of lignocellulose biomass described in step (1) is mineral matter degraded, the hydrolysis of hydro-thermal sulfur dioxide
One or more of combinations in method, acid steam explosion method and enzymatic isolation method.
Step (3) is specially：Ligno-cellulose hydrolysate pH is 3.5-5.5, and the lignin of addition is as detoxification reagent, wood
The quality of quality and ligno-cellulose hydrolysate volume are 0.5%-20%g/ml, then add saccharomyces cerevisiae, saccharomyces cerevisiae
The OD of cell density600For 0.2~20, ferment under the conditions of being 28-40 DEG C in temperature, fermentation time for 2-120 it is small when, production
Ethanol.
The quality of the lignin is preferably in a proportion of 1.0~10%g/ml with ligno-cellulose hydrolysate volume.
The OD of the cell density of the saccharomyces cerevisiae600It is worth and is：0.2~20, it is preferably 8~10.
The lignin is Bu Laosi (Brauns) native lignin (BNL), Nuo De (Nard) lignin, Beckman
(Bjorkman) lignin (MWL), cellulolytic enzyme lignin (CHL), ethanol lignin, coloured glaze guanidine-acetic acid lignin, phenol-lignin, lignin sulphur
Acid, alkali lignin (Kraft lignins), sulfidated lignin, chlorinated lignin, proflavine sulfate (Klason lignins), hydrochloric acid lignin, cuprammonium wood
One or more in element, periodates lignin.
The lignocellulosic comes from wheat stalk, maize straw or/and straw；The branches and leaves of lumbering generation, discarded wood
Head or/and sawdust；Sugar grass or/and withy journey or urban solid garbage, waste paper or/and bagasse.
The lignocellulosic from agricultural wastes, forestry waste, special energy crop or/and it is various containing fibre
Tie up the discarded object of element.
The agricultural wastes are wheat stalk, maize straw or/and straw.
Branches and leaves, discarded wood or/and the sawdust that the forestry waste produces for lumbering.
The special energy crop is sugar grass or/and withy journey.
The discarded object of the various containing celluloses is urban solid garbage, waste paper or/and bagasse.
The present invention provides a kind of new method of ligno-cellulose hydrolysate original position detoxification fermentation, this method is with absorption
The lignin of performance carries out the toxicant in ligno-cellulose hydrolysate for detoxification reagent the detoxification in situ in fermentation process,
So as to high-efficiency fermenting production alcohol fuel.Compared with the prior art, compared with existing fermentation of ligno-cellulose hydrolysate method,
Hydrolyzate of the present invention can be directly used for fermenting without detoxification treatment, simplifies production technology, reduces equipment investment；And detoxification reagent
It is of low cost, derive from a wealth of sources, it is easy to be recycled.
Brief description of the drawings
Fig. 1 is alkali lignin pair during the fermentation of ligno-cellulose hydrolysate producing and ethanol simulated in embodiment 1 and comparative example 1
The influence of concentration of alcohol；
Fig. 2 is alkali lignin pair during the fermentation of ligno-cellulose hydrolysate producing and ethanol simulated in embodiment 1 and comparative example 1
The influence of concentration of glucose；
Fig. 3 is alkali lignin pair during the fermentation of ligno-cellulose hydrolysate producing and ethanol simulated in embodiment 2 and comparative example 2
The influence of concentration of alcohol；
Fig. 4 is alkali lignin pair during the fermentation of ligno-cellulose hydrolysate producing and ethanol simulated in embodiment 2 and comparative example 2
The influence of concentration of glucose；
Fig. 5 is enzymolysis lignin during the fermentation of ligno-cellulose hydrolysate producing and ethanol simulated in embodiment 3 and comparative example 3
Influence to concentration of alcohol；
Fig. 6 is enzymolysis lignin during the fermentation of ligno-cellulose hydrolysate producing and ethanol simulated in embodiment 3 and comparative example 3
Influence to concentration of glucose；
Fig. 7 is that embodiment 4 and enzymolysis lignin during corn stalk hydrolysis fermentation producing and ethanol in comparative example 4 are dense to ethanol
The influence of degree；
Fig. 8 is embodiment 4 with enzymolysis lignin during corn stalk hydrolysis fermentation producing and ethanol in comparative example 4 to glucose
The influence of concentration.
Embodiment
With reference to specific embodiment, the present invention is described further, but protection scope of the present invention is from embodiment
Limitation, the content described in following embodiments and specification merely illustrates the principles of the invention, and is not departing from spirit of the invention
On the premise of scope, various changes and modifications of the present invention are possible, these changes and improvements both fall within this claimed hair
In bright scope.The scope of the present invention is defined by the appended claims and its equivalents.
Embodiment 1
In the present embodiment, the ligno-cellulose hydrolysate of simulation is made of glucose, the buffer solution that PH is 4.5；Measure mould
10 milliliters of the ligno-cellulose hydrolysate of plan, 0.7 gram of alkali lignin of addition, the cell density OD of saccharomyces cerevisiae600For 0.16, sending out
Fermentative production of ethanol under the conditions of when ferment temperature is 33 DEG C, fermentation time is 72 small.
Comparative example 1
The present embodiment is the comparative example of above-described embodiment 1.In the present embodiment except not adding lignin, remaining step
Suddenly it is identical with embodiment 1.10 milliliters of the ligno-cellulose hydrolysate of simulation is measured, adds the cell density of saccharomyces cerevisiae
OD600For 0.16, fermentation temperature be 33 DEG C, fermentation time be 72 small when under the conditions of fermentative production of ethanol.
After when fermentation 72 is small in above-described embodiment 1 and comparative example 1 glucose with concentration of alcohol as shown in Figure 1, 2.Have
Fig. 1,2 understand glucose solution ferment under the conditions of existing for alkali lignin 24 can complete fermentation generate ethanol, with comparative example phase
Compare, when fermentation termination shortening 24 is small.
Embodiment 2
In the present embodiment, the ligno-cellulose hydrolysate of simulation is by glucose, 2.0 grams/L phenol, the buffer solution that PH is 4.5
Composition；Measure 10 milliliters of the ligno-cellulose hydrolysate of simulation, 0.7 gram of alkali lignin of addition, the cell density OD of saccharomyces cerevisiae600
For 0.16, fermentation temperature be 33 DEG C, fermentation time be 72 small when under the conditions of fermentative production of ethanol.
Comparative example 2
The present embodiment is the comparative example of above-described embodiment 2.In the present embodiment except not adding lignin, remaining step
Suddenly it is identical with embodiment 2.10 milliliters of the ligno-cellulose hydrolysate of simulation is measured, adds the cell density of saccharomyces cerevisiae
OD600For 0.16, fermentation temperature be 33 DEG C, fermentation time be 72 small when under the conditions of fermentative production of ethanol.
After when fermentation 72 is small in above-described embodiment 2 and comparative example 2 glucose with concentration of alcohol as shown in Figure 3,4.Have
Fig. 3,4 understand that 2.0 grams/L phenol suppresses glucose fermentation, when fermentation 48 is small after only 18 g/l of ethanol produce；But when addition
After 0.7 gram of alkali lignin, glucose fermentation efficiency is obviously improved, fermentation 48 it is small when after concentration of alcohol significantly improve to 26 g/l.This
Absolutely prove that alkali lignin plays obvious detoxification in situ to 2.0 g/l of phenol.
Embodiment 3
In the present embodiment, the ligno-cellulose hydrolysate of simulation is by glucose, 2.0 grams/L phenol, the buffer solution that PH is 4.5
Composition；Measure 10 milliliters of the ligno-cellulose hydrolysate of simulation, 0.7 gram of addition cellulase hydrolysis lignin, the cell of saccharomyces cerevisiae
Density OD600For 0.16, fermentation temperature be 33 DEG C, fermentation time be 72 small when under the conditions of fermentative production of ethanol.
Comparative example 3
The present embodiment is the comparative example of above-described embodiment 3.In the present embodiment except not adding lignin, remaining step
Suddenly it is identical with embodiment 3.10 milliliters of the ligno-cellulose hydrolysate of simulation is measured, adds the cell density of saccharomyces cerevisiae
OD600For 0.16, fermentation temperature be 33 DEG C, fermentation time be 72 small when under the conditions of fermentative production of ethanol.
After when fermentation 72 is small in above-described embodiment 3 and comparative example 3 glucose with concentration of alcohol as shown in Figure 5,6.Have
Fig. 5,6 understand that 2.0 grams/L phenol suppresses glucose fermentation, when fermentation 48 is small after only 18 g/l of ethanol produce；But when addition
After 0.7 gram of cellulase hydrolysis lignin, glucose fermentation efficiency is obviously improved, fermentation 48 it is small when after concentration of alcohol significantly improve to 26
G/l.This has absolutely proved that cellulase hydrolysis lignin also plays obvious detoxification in situ to 2.0 g/l of phenol.
Embodiment 4
In the present embodiment, lignocellulose biomass is maize straw.Maize straw abolishes half fiber through steaming acid steam explosion
Cellulase hydrolysis is carried out after dimension element and obtains lignocellulosic hydrolysis slurries, residue remaining liq is then filtered to remove and ferment
To ethanol.Measure 10 milliliters of ligno-cellulose hydrolysate, 0.7 gram of addition cellulase hydrolysis lignin, the cell density of saccharomyces cerevisiae
OD600For 0.16, fermentation temperature be 33 DEG C, fermentation time be 72 small when under the conditions of fermentative production of ethanol.
After tested, following fermentation inhibitor is mainly contained in corn stalk hydrolysis：Phenol, guaiacol, vanillic aldehyde, fourth
Fragrant aldehyde, furfural, 5 hydroxymethyl furfural, benzoic acid.
Comparative example 4
The present embodiment is the comparative example of above-described embodiment 4.In the present embodiment except not adding lignin, remaining step
Suddenly it is identical with embodiment 4.10 milliliters of ligno-cellulose hydrolysate is measured, adds the cell density OD of saccharomyces cerevisiae600For
0.16, fermentation temperature be 33 DEG C, fermentation time be 72 small when under the conditions of fermentative production of ethanol.
Ethanol and concentration of glucose change curve be such as in system after everfermentation in above-described embodiment 4 and comparative example 4
Fig. 7, shown in 8.There are Fig. 7,8 to understand, when fermentation 96 is small after concentration of alcohol be 30 g/l, and without second in the zymotic fluid of detoxification
Alcohol content is only 3.9 g/l.This absolutely proved cellulase hydrolysis lignin to contain in ligno-cellulose hydrolysate phenol,
The toxic chemical of the suppression such as guaiacol, syringaldehyde, furfural, 5 hydroxymethyl furfural fermentation plays obvious detoxification in situ and makees
With.
Technical scheme is described in detail in all embodiments described above, it should be understood that the above is only
For the specific embodiment of the present invention, it is not intended to limit the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (12)
1. the detoxification fermentation process in situ of a kind of ligno-cellulose hydrolysate, it is characterised in that follow the steps below：
(1) the hydrolysis slurries that the lignocellulosic of pretreatment obtains after degraded；
(2) the hydrolysis slurries of acquisition are subjected to filtering and remove solid residue, obtain ligno-cellulose hydrolysate；
(3) and then into ligno-cellulose hydrolysate lignin, saccharomyces cerevisiae progress fermentation process are added, obtains alcohol fuel.
2. a kind of detoxification fermentation process in situ of ligno-cellulose hydrolysate described in accordance with the claim 1, it is characterized in that：It is described
The biodegrading process of step (1) lignocellulose biomass be mineral matter degraded, hydro-thermal sulfur dioxide Hydrolyze method, acid steam explosion method and
One or more of combinations in enzymatic isolation method.
A kind of 3. detoxification fermentation process in situ of ligno-cellulose hydrolysate described in accordance with the claim 1, it is characterised in that：Institute
The lignin stated is Bu Laosi native lignins, promise moral lignin, Beckman lignin, cellulose hydrolyzation lignin, ethanol lignin, dioxy
Own ring lignin, coloured glaze guanidine-acetic acid lignin, phenol-lignin, lignosulphonic acid, alkali lignin, sulfidated lignin, chlorinated lignin, proflavine sulfate, hydrochloric acid
One or more in lignin, cuprammonium lignin, periodates lignin.
A kind of 4. detoxification fermentation process in situ of ligno-cellulose hydrolysate described in accordance with the claim 1, it is characterised in that：Institute
The lignocellulosic stated is lignin in protist matter raw material, and the protist matter raw material is various trees, crops straw
One or several kinds of mixtures in stalk, farming industry byproduct, feces of livestock and poultry and energy crop etc., described kind of trees are
Cork or/and hardwood.
A kind of 5. detoxification fermentation process in situ of ligno-cellulose hydrolysate described in accordance with the claim 1, it is characterised in that：Institute
The lignocellulosic stated is discarded from agricultural wastes, forestry waste, special energy crop or/and various containing celluloses
Thing.
6. according to a kind of detoxification fermentation process in situ of ligno-cellulose hydrolysate described in claim 5, it is characterised in that：Institute
It is wheat stalk, maize straw or/and straw to state agricultural wastes.
7. according to a kind of detoxification fermentation process in situ of ligno-cellulose hydrolysate described in claim 5, it is characterised in that：Institute
State branches and leaves, discarded wood or/and sawdust that forestry waste produces for lumbering.
8. according to a kind of detoxification fermentation process in situ of ligno-cellulose hydrolysate described in claim 5, it is characterised in that：Institute
It is sugar grass or/and withy journey to state special energy crop.
9. according to a kind of detoxification fermentation process in situ of ligno-cellulose hydrolysate described in claim 5, it is characterised in that：Institute
The discarded object for stating various containing celluloses is urban solid garbage, waste paper or/and bagasse.
A kind of 10. detoxification fermentation process in situ of ligno-cellulose hydrolysate described in accordance with the claim 1, it is characterised in that：
The ratio between the lignin quality and the volume of ligno-cellulose hydrolysate are 0.5-20%g/ml.
A kind of 11. detoxification fermentation process in situ of ligno-cellulose hydrolysate described in accordance with the claim 1, it is characterised in that：
The OD of the cell density of saccharomyces cerevisiae described in step (3)600It is worth and is：0.2~20, fermentation temperature for 28-40 DEG C, fermentation when
Between for 2-120 it is small when.
12. according to a kind of detoxification fermentation process in situ of ligno-cellulose hydrolysate described in claim 11, it is characterised in that：
The OD of the cell density of the saccharomyces cerevisiae600Value is preferably 8~10.
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Cited By (1)
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