CN1944658A - Method for producing cellulose alcohol using corncob processing leftover by fermenting - Google Patents
Method for producing cellulose alcohol using corncob processing leftover by fermenting Download PDFInfo
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- CN1944658A CN1944658A CNA200610131965XA CN200610131965A CN1944658A CN 1944658 A CN1944658 A CN 1944658A CN A200610131965X A CNA200610131965X A CN A200610131965XA CN 200610131965 A CN200610131965 A CN 200610131965A CN 1944658 A CN1944658 A CN 1944658A
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- slag
- alcohol
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- liquid
- corn cob
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 158
- 229920002678 cellulose Polymers 0.000 title claims abstract description 44
- 239000001913 cellulose Substances 0.000 title claims abstract description 44
- 238000004519 manufacturing process Methods 0.000 title abstract description 16
- 239000007788 liquid Substances 0.000 claims abstract description 100
- 238000000855 fermentation Methods 0.000 claims abstract description 70
- 230000004151 fermentation Effects 0.000 claims abstract description 70
- 240000008042 Zea mays Species 0.000 claims abstract description 34
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 34
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 34
- 235000005822 corn Nutrition 0.000 claims abstract description 34
- 108010059892 Cellulase Proteins 0.000 claims abstract description 25
- 229940106157 cellulase Drugs 0.000 claims abstract description 25
- 241000894006 Bacteria Species 0.000 claims abstract description 21
- 239000002893 slag Substances 0.000 claims description 127
- 235000019441 ethanol Nutrition 0.000 claims description 116
- HYBBIBNJHNGZAN-UHFFFAOYSA-N furfural Chemical compound O=CC1=CC=CO1 HYBBIBNJHNGZAN-UHFFFAOYSA-N 0.000 claims description 76
- 108090000790 Enzymes Proteins 0.000 claims description 65
- 102000004190 Enzymes Human genes 0.000 claims description 65
- 229940088598 enzyme Drugs 0.000 claims description 65
- 239000000203 mixture Substances 0.000 claims description 51
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 claims description 50
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 claims description 50
- 239000000811 xylitol Substances 0.000 claims description 50
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 claims description 50
- 229960002675 xylitol Drugs 0.000 claims description 50
- 235000010447 xylitol Nutrition 0.000 claims description 50
- 238000000034 method Methods 0.000 claims description 49
- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 claims description 47
- 230000001580 bacterial effect Effects 0.000 claims description 44
- 235000010980 cellulose Nutrition 0.000 claims description 42
- 238000002360 preparation method Methods 0.000 claims description 36
- 238000012262 fermentative production Methods 0.000 claims description 31
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 27
- 239000010909 process residue Substances 0.000 claims description 26
- 238000002156 mixing Methods 0.000 claims description 21
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 20
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 20
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 20
- 235000015099 wheat brans Nutrition 0.000 claims description 20
- 229920000168 Microcrystalline cellulose Polymers 0.000 claims description 11
- 238000010564 aerobic fermentation Methods 0.000 claims description 11
- 238000011534 incubation Methods 0.000 claims description 11
- 235000019813 microcrystalline cellulose Nutrition 0.000 claims description 11
- 239000008108 microcrystalline cellulose Substances 0.000 claims description 11
- 229940016286 microcrystalline cellulose Drugs 0.000 claims description 11
- 241001304120 Trichoderma pseudokoningii Species 0.000 claims description 10
- 239000002023 wood Substances 0.000 claims description 10
- 229920001817 Agar Polymers 0.000 claims description 9
- 239000001888 Peptone Substances 0.000 claims description 9
- 108010080698 Peptones Proteins 0.000 claims description 9
- 239000008272 agar Substances 0.000 claims description 9
- 230000003321 amplification Effects 0.000 claims description 9
- 108010047754 beta-Glucosidase Proteins 0.000 claims description 9
- 102000006995 beta-Glucosidase Human genes 0.000 claims description 9
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 9
- 235000019319 peptone Nutrition 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 239000006052 feed supplement Substances 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 7
- 239000000047 product Substances 0.000 claims description 5
- 238000012807 shake-flask culturing Methods 0.000 claims description 4
- 239000002440 industrial waste Substances 0.000 abstract description 3
- 238000009776 industrial production Methods 0.000 abstract description 2
- 230000001360 synchronised effect Effects 0.000 abstract 1
- 229960004756 ethanol Drugs 0.000 description 31
- 239000000463 material Substances 0.000 description 7
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- 239000002699 waste material Substances 0.000 description 5
- 239000002994 raw material Substances 0.000 description 4
- 238000003912 environmental pollution Methods 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 235000019341 magnesium sulphate Nutrition 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000007836 KH2PO4 Substances 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 229920005610 lignin Polymers 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 108010084185 Cellulases Proteins 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- HEBKCHPVOIAQTA-NGQZWQHPSA-N d-xylitol Chemical compound OC[C@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-NGQZWQHPSA-N 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 230000005619 thermoelectricity Effects 0.000 description 1
- 229960003487 xylose Drugs 0.000 description 1
Images
Classifications
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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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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses fermentation process of producing cellulose alcohol with corn cob processing leftover. The fermentation process includes bacteria sorting, preparing liquid fermentation bacteria, preparing coarse cellulase liquid, synchronous anaerobic saccharification and fermentation to produce alcohol, purifying alcohol and other steps. The present invention has industrial waste utilized, so that it has low production cost, high stability and environment friendship and is suitable for industrial production.
Description
Technical field
The present invention relates to a kind of method of production of cellulose alcohol, relate in particular to a kind of corn cob process residues that utilizes---xylose residue, Xylitol slag, furfural slag fermentative production cellulase solution, and then the method for enzymolysis xylose residue, Xylitol slag or furfural slag production alcohol, belong to the microbial fermentation production field.
Background technology
Fermentative Production alcohol is raw material with cereal, potato class and molasses mainly at present, its complex manufacturing, and the cost height exists with the people and strives the problem that grain is striven ground.From opening up the new energy and the consideration of curbing environmental pollution, be subject to people's attention day by day with waste cellulose class resource production Study of Alcohol.Although the utilization with waste cellulose class resource is reported to some extent, but, utilize the corn cob process residues---xylose residue, Xylitol slag, furfural slag liquid submerged fermentation production of cellulose enzyme liquid, and then enzymolysis xylose residue, Xylitol slag or furfural slag are produced the method for alcohol, by retrieval, yet there are no report.
Summary of the invention
At the deficiencies in the prior art, the problem to be solved in the present invention is: adopt modern biotechnology, utilize the industrial waste fibrous matter, particularly the corn cob process residues---xylose residue, Xylitol slag, furfural slag are produced high active cellulase liquid by liquid submerged fermentation, and then utilize corn cob process residues such as cellulase solution enzymolysis xylose residue, Xylitol slag, furfural slag to produce the method for alcohol.
The method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol of the present invention, form by following steps:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:, under aseptic condition, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.0～1.2 * 10/ml with step (1) described bacterial strain;
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the bacterial classification proliferated culture medium, its composition and preparation are: xylose residue or Xylitol slag or furfural slag or its are mixed slag, wheat bran, peptone, ammonium sulfate, water by weight 1: 2: 0.3: 0.2: 100 ratio cooperates, the pH nature;
Described shake-flask culture condition is 28 ℃-33 ℃ of temperature, shaking speed 150-180r/min, incubation time 30-60 hour; First class seed pot, secondary seed jar culture condition are 28 ℃-33 ℃ of temperature, tank pressure 0.5Kg/cm
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute the meter 1: 0.8-1.0, mixing speed 100-150r/min, aerobic cultivation 30-60 hour;
(3) Mierocrystalline cellulose crude enzyme liquid preparation: use and produce the enzyme substratum, pack in the fermentor tank with the amount of coefficient 70-80%,, add the described liquid fermenting bacterial classification of 10% step (2) again with the weight percent meter of charge amount, with 28 ℃-33 ℃ of culture temperature, tank pressure 0.6～1kg/cm
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 0.8～1.0, the condition of mixing speed 100～150r/min aerobic fermentation 80-136 hour, makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
Wherein: described product enzyme substratum, its composition and preparation method are: xylose residue or Xylitol slag or furfural slag or its are mixed slag: wheat bran: water is with 2-5: 2-5: 100 ratio cooperates; Again by weight percentage, add microcrystalline cellulose 0.4-0.8%, ammonium sulfate 0.2-0.5%, KH
2PO
40.2-0.4%, MgSO
40.04-0.06%, pH 5.0-6.0;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get crude enzyme liquid that step (3) makes and corn cob process residues-xylose residue or Xylitol slag or furfural slag or its and mix slag and mix in ethanol fermentation tank with the ratio of the dried slag of enzyme amount 10FPU/g, the distillery yeast that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 28 ℃-35 ℃, and the pH value is 4-5; Ferment 12-48 hour the time,, mend or mend in batches above-mentioned xylose residue or Xylitol slag or furfural slag or its again and mix the xylose residue of slag gross dry weight ratio 40%～60% or Xylitol slag or furfural slag or its and mix slag according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 60-72 hour, fermentation is finished;
Wherein: described distillery yeast is the yeast mash that cell count reaches 100,000,000/ml, bud ratio 15-30%;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills, and promptly gets alcohol.
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: described xylose residue of step (3) or Xylitol slag or furfural slag or its mix slag: wheat bran: the weight ratio of water preferably 3: 3: 100.
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: the described microcrystalline cellulose of step (3) is 0.5-0.7% preferably, and ammonium sulfate is 0.3-0.4% preferably, KH
2PO
40.3-0.4% preferably, MgSO
40.05-0.06% preferably, the preferred 5.4-5.8 of pH.
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: preferably 30 ℃-33 ℃ of step (2) or the described culture temperature of step (3).
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: step (2) or (3) described air flow are with fermentating liquid volume m
3/ volume of air m
3Minute the meter be preferably 1: 0.9.
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: preferably 100-136 hour step (3) described aerobic fermentation time.
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: the described leavening temperature of step (4) is preferably 30 ℃-33 ℃, and the pH value is preferably 4.3～4.8.
Wherein: the described leavening temperature of step (4) most preferably is 30 ℃-32 ℃, and the pH value most preferably is 4.5～4.6.
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: step (2), (3) or (4) described mixing slag are meant the mixture that xylose residue and Xylitol slag or furfural slag or its three cooperate with any part by weight.
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: step (4) is described to be meant that according to the karusen ethanol concn ethanol concn is when volume percent 8% is following; Described feed supplement amount determines that according to the karusen ethanol concn described karusen ethanol concn is low more, and feed supplement number of times and feed supplement amount are many more, and common feed supplement number of times is 1～2 time.
In the above-mentioned method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol: the index that the described fermentation of step (4) is finished is that the karusen ethanol concn reaches volume percent more than 8%.
Utilize method of the present invention can successfully utilize the industrial waste fibrous matter, corn cob process residues particularly---xylose residue, Xylitol slag, furfural slag are produced high active cellulase liquid by liquid submerged fermentation, and then residues such as the described xylose residue of enzymolysis, Xylitol slag, furfural slag make alcohol; Make the alcohol raw material the yield of liquor and reach more than 20%, production cost is lower than grain alcohol, and the waste residue after the fermentation can be made fuel, has realized industrial residue is turned waste into wealth, and has also solved problem of environmental pollution simultaneously, has great application value.
Method of the present invention has following characteristics:
1, the employed raw material of the production of cellulase and Alcohol Production is useless the fibrous matter---xylose residue of industry, Xylitol slag or furfural slag, at present corn cob processing wood sugar, Xylitol, xylo-oligosaccharide, on the ripe basis of applying of series product such as furfural, corn cob process residues---xylose residue, the Xylitol slag, furfural slag is easy to concentrate and collect, and, corn cob and process residues thereof are through last product sequence processing treatment mistake, more help the decomposition utilization of microorganism, can induce a large amount of cellulases synthetic, reduce the raw materials pretreatment operation, reduced production cost.
2, to have adopted plan Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCCNo3.3002 of seed selection be bacterial classification to method of the present invention, liquid submerged fermentation, and the production stability height, pollution microbes is not convenient to large-scale industrial production.Residue main component after the fermentation is a lignin, and more former xylose residue of calorific value or Xylitol slag or furfural slag improve, but burning boiler, for Alcohol Production provides thermoelectric; Utilize residual lignin burning coproduction thermoelectricity, the three class main components that constitute corn cob are converted into the complete biorefinery flow process of variant production respectively, realized industrial residue is turned waste into wealth, also solved problem of environmental pollution simultaneously, had great society and economic worth.
3, technology of alcohol has adopted simultaneous saccharification and fermentation technology, has overcome the inhibition to cellulase of carbohydrate and other product, and has simplified the complicated technology of first saccharification secondary fermentation.
4, adopt the measure of middle feed supplement in the technology of alcohol, improved the ethanol concn of fermentation liquid, reduced the production cost of back extraction process.
Description of drawings
Fig. 1: the process flow sheet that utilizes corn cob process residues fermentative production Mierocrystalline cellulose alcohol.
Embodiment
Embodiment 1
Utilize the method for xylose residue fermentative production Mierocrystalline cellulose alcohol, form by following steps:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:,, under 30 ℃ of conditions, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.1 * 10/ml aseptic with step (1) described bacterial strain;
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the bacterial classification proliferated culture medium, its composition and preparation method are: with xylose residue, wheat bran, peptone, ammonium sulfate, water were by weight 1: 2: 0.3: 0.2: 100 ratio cooperates, the PH nature.
30 ℃ of described shake-flask culture temperature, shaking speed 160r/min, incubation time 30 hours, first class seed pot, secondary seed jar: 30 ℃ of culture temperature, tank pressure 0.5kg/cm
2, air flow is with fermentating liquid volume m
3/ gas volume m
3Minute the meter, 1: 0.9, mixing speed 130-150r/min, incubation time, first class seed pot 35 hours, secondary seed jar 45 hours.
(3) Mierocrystalline cellulose crude enzyme liquid preparation: employed substratum is to produce the enzyme substratum, and its composition and preparation method are: cooperate with 3: 3: 100 ratio with weight percent xylose residue, wheat bran, water; Again by weight percentage, add microcrystalline cellulose 06%, ammonium sulfate 0.4%, KH2PO4 0.3%, and MgSO4 0.05%, pH5.6, pack in the fermentor tank with the amount of coefficient 75%,, add the described liquid fermenting bacterial classification of 10% step (2) again with the weight percent meter of charge amount, with 30 ℃ of culture temperature, tank pressure 0.8kg/cm
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 0.9, the condition of mixing speed 120r/min, aerobic fermentation 108 hours makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get the crude enzyme liquid that step (3) makes and mix (enzyme liquid: material=weight ratio 1.5: 1) with xylose residue with the ratio of the dried slag of enzyme amount 10FPU/g in ethanol fermentation tank, the distillery yeast (yeast count reaches the distiller's yeast mash of 100,000,000/ml, bud ratio 15-30%) that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 30 ℃, and the pH value is 4.6; Ferment 24 hours the time,, mend the xylose residue of above-mentioned xylose residue gross dry weight ratio 50% again according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 65-72 hour, fermentation is finished;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills with ordinary method, promptly gets alcohol.
Embodiment 2
Utilize the method for Xylitol slag fermentative production Mierocrystalline cellulose alcohol, form by following steps:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:,, under 28 ℃ of conditions, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.0 * 10/ml aseptic with step (1) described bacterial strain;
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the bacterial classification proliferated culture medium, its composition and preparation method are: with Xylitol slag, wheat bran, peptone, ammonium sulfate, water by weight 1: 2: 0.3: 0.2: 100 ratio cooperates, the PH nature.
Described 28 ℃ of bottle, the culture temperature of shaking; Shaking speed 150-160r/min, 35 hours shaking culture time first class seed pot, secondary seed jar culture temperature are 28 ℃; Tank pressure 0.5Kg/cm
2, air flow fermentating liquid volume m
3/ volume of air m
3Minute the meter, 1: 0.8, mixing speed 100-110r/min, incubation time, first class seed pot 40 hours, secondary seed jar 50 hours.
(3) Mierocrystalline cellulose crude enzyme liquid preparation: the substratum that uses is to produce the enzyme substratum; Its composition and preparation method are: Xylitol slag, wheat bran, water are cooperated by weight the ratio with 2.5: 2.5: 100; Again by weight percentage, add microcrystalline cellulose 0.4%, ammonium sulfate 0.2%, KH
2PO4 0.2%, and MgSO4 0.04%, pH5.0; Pack in the fermentor tank with the amount of coefficient 70%,, add the described liquid fermenting bacterial classification of 10% step (2), with 28 ℃ of culture temperature, tank pressure 0.6kg/cm again with the weight percent meter of charge amount
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 0.8, the condition of mixing speed 100r/min, aerobic fermentation 86 hours makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get the crude enzyme liquid that step (3) makes and mix (enzyme liquid: material=weight ratio 1: 1) with the Xylitol slag with the ratio of the dried slag of enzyme amount 10FPU/g in ethanol fermentation tank, the distillery yeast (yeast count reaches the distiller's yeast mash of 100,000,000/ml, bud ratio 15-30%) that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 28 ℃, and the pH value is 4.3; Ferment 12 hours the time,, mend the Xylitol slag of above-mentioned Xylitol slag gross dry weight ratio 40% more in two batches according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 60-66 hour, fermentation is finished;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills with ordinary method, promptly gets alcohol.
Embodiment 3
Utilize the method for xylose residue fermentative production Mierocrystalline cellulose alcohol, form by following steps:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:,, under 33 ℃ of conditions, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.2 * 10/ml aseptic with step (1) described bacterial strain;
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the bacterial classification proliferated culture medium, its composition and preparation method are: with xylose residue, wheat bran, peptone, ammonium sulfate, water by weight 1: 2: 0.3: 0.2: 100 ratio cooperates, the pH nature;
Described 33 ℃ of bottle, the culture temperature of shaking; Shaking speed 170-180r/min, 38 hours shaking culture time first class seed pot, secondary seed jar culture temperature are 33 ℃; Tank pressure 0.5Kg/cm
2, air flow fermentating liquid volume m
3/ volume of air m
3Minute the meter, 1: 1.0, mixing speed 140-150r/min, incubation time, first class seed pot 46 hours, secondary seed jar 60 hours.
(3) Mierocrystalline cellulose crude enzyme liquid preparation: employed substratum is to produce the enzyme substratum, and its composition and preparation method are: xylose residue, wheat bran, water cooperate with 5: 5: 100 ratio; Again by weight percentage, add microcrystalline cellulose 0.8%, ammonium sulfate 0.5%, KH
2PO
40.4%, MgSO
40.06%, pH 6.0; Pack in the fermentor tank with the amount of coefficient 80%,, add the described liquid fermenting bacterial classification of 10% step (2), with 33 ℃ of culture temperature, tank pressure 1kg/cm again with the weight percent meter of charge amount
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 1.0, the condition of mixing speed 150r/min, aerobic fermentation 120 hours makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get the crude enzyme liquid that step (3) makes and mix (enzyme liquid: material=weight ratio 2: 1) with xylose residue with the ratio of the dried slag of enzyme amount 10FPU/g in ethanol fermentation tank, the distillery yeast (yeast count reaches the distiller's yeast mash of 100,000,000/ml, bud ratio 15-30%) that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 33 ℃, and the pH value is 5.0; Ferment 48 hours the time,, mend the xylose residue of above-mentioned xylose residue gross dry weight ratio 60% more in three batches according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 70-72 hour, fermentation is finished;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills with ordinary method, promptly gets alcohol.
Embodiment 4
Utilize the method for xylose residue and Xylitol slag balanced mix slag fermentative production Mierocrystalline cellulose alcohol, form by following steps:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:,, under 30 ℃ of conditions, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.1 * 10/ml aseptic with step (1) described bacterial strain;
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the growing microorganism substratum, its composition and preparation method are: with xylose residue and Xylitol slag balanced mix slag, wheat bran, peptone, ammonium sulfate, water by weight 1: 2: 0.3: 0.2: 100 ratio cooperates, and PH is naturally;
Described shaking 30 ℃ of bottle, culture temperature, shaking speed 160-170r/min, 30 hours shaking culture time; First class seed pot, secondary seed jar culture temperature are 30 ℃; Tank pressure 0.5Kg/cm
2, air flow fermentating liquid volume m
3/ volume of air m
3Minute the meter, 1: 0.9, mixing speed 120-130r/min, incubation time, first class seed pot 35 hours, secondary seed jar 45 hours.
(3) Mierocrystalline cellulose crude enzyme liquid preparation: employed substratum is to produce the enzyme substratum, and its composition and preparation method are: cooperate by weight 3: 3: 100 ratio with xylose residue and Xylitol slag balanced mix slag, wheat bran, water; Again by weight percentage, add microcrystalline cellulose 0.6%, ammonium sulfate 0.4%, KH
2PO
40.3%, MgSO
40.05%, pH 5.6; Pack in the fermentor tank with the amount of coefficient 75%,, add the described liquid fermenting bacterial classification of 10% step (2), with 30 ℃ of culture temperature, tank pressure 0.8kg/cm again with the weight percent meter of charge amount
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 0.9, the condition of mixing speed 120r/min, aerobic fermentation 108 hours makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get the crude enzyme liquid that step (3) makes and in ethanol fermentation tank mix (enzyme liquid: material=weight ratio 1.5: 1) with Xylitol slag balanced mix slag with the ratio of the dried slag of enzyme amount 10FPU/g with xylose residue, the distillery yeast (yeast count reaches the distiller's yeast mash of 100,000,000/ml, bud ratio 15-30%) that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 30 ℃, and the pH value is 4.6; Ferment 24 hours the time,, mend the xylose residue and the Xylitol slag balanced mix slag of above-mentioned xylose residue and Xylitol slag balanced mix slag gross dry weight ratio 50% again according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 65-72 hour, fermentation is finished;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills with ordinary method, promptly gets alcohol.
Embodiment 5
Utilize xylose residue and Xylitol slag to mix the method for slag fermentative production Mierocrystalline cellulose alcohol at 3: 2, form by following steps:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:,, under 32 ℃ of conditions, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.0 * 10/ml aseptic with step (1) described bacterial strain; (bacterium number should with the OD value representation).
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the bacterial classification proliferated culture medium, its composition and preparation method are: xylose residue and Xylitol slag are mixed slag, wheat bran, peptone, ammonium sulfate, water by weight 1: 2: 0.3 at 3: 2: 0.2: 100 ratio cooperates, the PH nature;
Described shaking 32 ℃ of bottle, culture temperature, shaking speed 160-170r/min, 33 hours shaking culture time; First class seed pot, secondary seed jar culture temperature are 32 ℃; Tank pressure 0.5Kg/cm
2, air flow fermentating liquid volume m
3/ volume of air m
3Minute the meter, 1: 1.0, mixing speed 110-120r/min, incubation time, first class seed pot 38 hours, secondary seed jar 48 hours.
(3) Mierocrystalline cellulose crude enzyme liquid preparation: employed substratum is to produce the enzyme substratum, and its composition and preparation method are: xylose residue and Xylitol slag mixing in 3: 2 slag, wheat bran, water are cooperated by weight 3: 3: 100 ratio; Again by weight percentage, add microcrystalline cellulose 0.6%, ammonium sulfate 0.5%, KH
2PO
40.4%, MgSO
40.06%, pH 5.8;
Pack in the fermentor tank with the amount of coefficient 75%,, add the described liquid fermenting bacterial classification of 10% step (2), with 32 ℃ of culture temperature, tank pressure 1.0kg/cm again with the weight percent meter of charge amount
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 1.0, the condition of mixing speed 130r/min, aerobic fermentation 118 hours makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get crude enzyme liquid that step (3) makes and mix slag at 3: 2 with xylose residue and Xylitol slag and mix (enzyme liquid: material=weight ratio 1.3: 1) with the ratio of the dried slag of enzyme amount 10FPU/g in ethanol fermentation tank, the distillery yeast (yeast count reaches the distiller's yeast mash of 100,000,000/ml, bud ratio 15-30%) that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 32 ℃, and the pH value is 4.8; Ferment 28 hours the time,, mend above-mentioned xylose residue more in two batches and mix slag at 3: 2 with the Xylitol slag with the xylose residue that the Xylitol slag mixes slag gross dry weight ratio 50% at 3: 2 according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 68-72 hour, fermentation is finished;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills with ordinary method, promptly gets alcohol.
Embodiment 6
Utilize the method for furfural slag fermentative production Mierocrystalline cellulose alcohol, form by following steps:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:,, under 30 ℃ of conditions, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.1 * 10/ml aseptic with step (1) described bacterial strain;
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the bacterial classification proliferated culture medium, its composition and preparation method are: with furfural slag, wheat bran, peptone, ammonium sulfate, water were by weight 1: 2: 0.3: 0.2: 100 ratio cooperates, the PH nature.
30 ℃ of described shake-flask culture temperature, shaking speed 160r/min, incubation time 30 hours, first class seed pot, secondary seed jar: 30 ℃ of culture temperature, tank pressure 0.5kg/cm
2, air flow is with fermentating liquid volume m
3/ gas volume m
3Minute the meter, 1: 0.9, mixing speed 130-150r/min, incubation time, first class seed pot 35 hours, secondary seed jar 45 hours.
(3) Mierocrystalline cellulose crude enzyme liquid preparation: employed substratum is to produce the enzyme substratum, and its composition and preparation method are:
Cooperate with 3: 3: 100 ratio with weight percent furfural slag, wheat bran, water; Again by weight percentage, add microcrystalline cellulose 06%, ammonium sulfate 0.4%, KH2PO4 0.3%, and MgSO4 0.05%, pH5.6, pack in the fermentor tank with the amount of coefficient 75%,, add the described liquid fermenting bacterial classification of 10% step (2) again with the weight percent meter of charge amount, with 30 ℃ of culture temperature, tank pressure 0.8kg/cm
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 0.9, the condition of mixing speed 120r/min, aerobic fermentation 108 hours makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get the crude enzyme liquid that step (3) makes and mix (enzyme liquid: material=weight ratio 1.5: 1) with furfural slag with the ratio of the dried slag of enzyme amount 10FPU/g in ethanol fermentation tank, the distillery yeast (yeast count reaches the distiller's yeast mash of 100,000,000/ml, bud ratio 15-30%) that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 30 ℃, and the pH value is 4.6; Ferment 24 hours the time,, mend the furfural slag of above-mentioned furfural slag gross dry weight ratio 50% again according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 65-72 hour, fermentation is finished;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills with ordinary method, promptly gets alcohol.
Embodiment 7
Utilize the method for xylose residue and Xylitol slag, furfural slag balanced mix slag fermentative production Mierocrystalline cellulose alcohol, form by following steps:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:,, under 30 ℃ of conditions, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.1 * 10/ml aseptic with step (1) described bacterial strain;
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the growing microorganism substratum, its composition and preparation method are: with xylose residue and Xylitol slag, furfural slag balanced mix slag, wheat bran, peptone, ammonium sulfate, water by weight 1: 2: 0.3: 0.2: 100 ratio cooperates, and pH is naturally;
Described shaking 30 ℃ of bottle, culture temperature, shaking speed 160-170r/min, 30 hours shaking culture time; First class seed pot, secondary seed jar culture temperature are 30 ℃; Tank pressure 0.5Kg/cm
2, air flow fermentating liquid volume m
3/ volume of air m
3Minute the meter, 1: 0.9, mixing speed 120-130r/min, incubation time, first class seed pot 35 hours, secondary seed jar 45 hours.
(3) Mierocrystalline cellulose crude enzyme liquid preparation: employed substratum is to produce the enzyme substratum, and its composition and preparation method are: cooperate by weight 3: 3: 100 ratio with xylose residue and Xylitol slag, furfural slag balanced mix slag, wheat bran, water; Again by weight percentage, add microcrystalline cellulose 0.6%, ammonium sulfate 0.4%, KH
2PO
40.3%, MgSO
40.05%, pH 5.6; Pack in the fermentor tank with the amount of coefficient 75%,, add the described liquid fermenting bacterial classification of 10% step (2), with 30 ℃ of culture temperature, tank pressure 0.8kg/cm again with the weight percent meter of charge amount
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 0.9, the condition of mixing speed 120r/min, aerobic fermentation 108 hours makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get the crude enzyme liquid that step (3) makes and in ethanol fermentation tank mix (enzyme liquid: material=weight ratio 1.5: 1) with Xylitol slag, furfural slag balanced mix slag with the ratio of the dried slag of enzyme amount 10FPU/g with xylose residue, the distillery yeast (yeast count reaches the distiller's yeast mash of 100,000,000/ml, bud ratio 15-30%) that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 30 ℃, and the pH value is 4.6; Ferment 24 hours the time,, mend the xylose residue of above-mentioned xylose residue and Xylitol slag, furfural slag balanced mix slag gross dry weight ratio 50% and Xylitol slag, furfural slag balanced mix slag again according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 65-72 hour, fermentation is finished;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills with ordinary method, promptly gets alcohol.
Claims (10)
1. method of utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol, form by following step:
(1) bacterial classification is selected: select for use cellulase producing bacteria to intend Kang Shi wood enzyme (Trichoderma Pseudokoningii) CGMCC No3.3002;
(2) liquid fermenting strain preparation:, under aseptic condition, through the test tube slant, shake bottle, first class seed pot, secondary seed jar amplification culture step by step, make the liquid fermenting bacterial classification that the bacterium number reaches OD value 1.0～1.2 * 10/ml with step (1) described bacterial strain;
Wherein:
Described slant culture based component is by weight percentage: 10% bran water adds 2% agar powder;
The described substratum that shakes bottle, first class seed pot, the use of secondary seed jar is the bacterial classification proliferated culture medium, its composition and preparation are: xylose residue or Xylitol slag or furfural slag or its are mixed slag, wheat bran, peptone, ammonium sulfate, water by weight 1: 2: 0.3: 0.2: 100 ratio cooperates, the pH nature;
Described shake-flask culture condition is 28 ℃-33 ℃ of temperature, shaking speed 150-180r/min, incubation time 30-60 hour; First class seed pot, secondary seed jar culture condition are 28 ℃-33 ℃ of temperature, tank pressure 0.5Kg/cm
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute the meter 1: 0.8-1.0, mixing speed 100-150r/min, aerobic cultivation 30-60 hour;
(3) Mierocrystalline cellulose crude enzyme liquid preparation: use and produce the enzyme substratum, pack in the fermentor tank with the amount of coefficient 70-80%,, add the described liquid fermenting bacterial classification of 10% step (2) again with the weight percent meter of charge amount, with 28 ℃-33 ℃ of culture temperature, tank pressure 0.6～1kg/cm
2, air flow is with fermentating liquid volume m
3/ volume of air m
3Minute meter 1: 0.8～1.0, the condition of mixing speed 100～150r/min aerobic fermentation 80-136 hour, makes the crude enzyme liquid of high active cellulase and beta-glucosidase;
Wherein: described product enzyme substratum, its composition and preparation method are: xylose residue or Xylitol slag or furfural slag or its are mixed slag: wheat bran: water is with 2-5: 2-5: 100 ratio cooperates; Again by weight percentage, add microcrystalline cellulose 0.4-0.8%, ammonium sulfate 0.2-0.5%, KH
2PO
40.2-0.4%, MgSO
40.04-0.06%, pH 5.0-6.0;
(4) the anaerobism simultaneous saccharification and fermentation is produced alcohol: get crude enzyme liquid that step (3) makes and corn cob process residues-xylose residue or Xylitol slag or furfural slag or its and mix slag and mix in ethanol fermentation tank with the ratio of the dried slag of enzyme amount 10FPU/g, the distillery yeast that adds total liquid weight 10% again, the anaerobism simultaneous saccharification and fermentation, leavening temperature is 28 ℃-35 ℃, and the pH value is 4-5; Ferment 12-48 hour the time,, mend or mend in batches above-mentioned xylose residue or Xylitol slag or furfural slag or its again and mix the xylose residue of slag gross dry weight ratio 40%～60% or Xylitol slag or furfural slag or its and mix slag according to the karusen ethanol concn; So that reach volume percent more than 8% in the karusen ethanol concn of fermentation time during to 60-72 hour, fermentation is finished;
Wherein: described distillery yeast is the yeast mash that cell count reaches 100,000,000/ml, bud ratio 15-30%;
(5) alcohol purifying: the alcohol mash after the fermentation that step (4) is made distills, and promptly gets alcohol.
2. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 1 is characterized in that: described xylose residue of step (3) or Xylitol slag or furfural slag or its mix slag: wheat bran: the weight ratio of water is 3: 3: 100.
3. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 1 is characterized in that: the described microcrystalline cellulose of step (3) is 0.5-0.7%, and ammonium sulfate is 0.3-0.4%, KH
2PO
4Be 0.3-0.4%, MgSO
4Be 0.05-0.06%, natural pH 5.4-5.8.
4. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 1 is characterized in that: step (2) or the described culture temperature of step (3) are 30 ℃-33 ℃.
5. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 1 is characterized in that: step (2) or (3) described air flow are with fermentating liquid volume m
3/ volume of air m
3Minute count 1: 0.9.
6. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 1 is characterized in that: the described aerobic fermentation time of step (3) is 100-136 hour.
7. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 1 is characterized in that: the described leavening temperature of step (4) is 30 ℃-33 ℃, and the pH value is 4.3～4.8.
8. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 7 is characterized in that: step (2), (3) or (4) described mixing slag are meant the mixture that cooperates with any part by weight between xylose residue and Xylitol slag or furfural slag or its three.
9. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 1 is characterized in that: step (4) is described to be meant that according to the karusen ethanol concn ethanol concn is when volume percent 8% is following; Described feed supplement amount determines that according to the karusen ethanol concn described karusen ethanol concn is low more, and feed supplement number of times and feed supplement amount are many more.
10. the method for utilizing corn cob process residues fermentative production Mierocrystalline cellulose alcohol as claimed in claim 1 is characterized in that: the index that the described fermentation of step (4) is finished is that the karusen ethanol concn reaches volume percent more than 8%.
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