CN109913507A - Synchronize the lignocellulosic method for refining for carrying out pretreatment and enzyme hydrolysis - Google Patents
Synchronize the lignocellulosic method for refining for carrying out pretreatment and enzyme hydrolysis Download PDFInfo
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- CN109913507A CN109913507A CN201910209583.1A CN201910209583A CN109913507A CN 109913507 A CN109913507 A CN 109913507A CN 201910209583 A CN201910209583 A CN 201910209583A CN 109913507 A CN109913507 A CN 109913507A
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- lignocellulosic
- urea
- enzyme hydrolysis
- refining
- hydrolase
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- 238000000034 method Methods 0.000 title claims abstract description 36
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 35
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 35
- 230000007062 hydrolysis Effects 0.000 title claims abstract description 31
- 238000006460 hydrolysis reaction Methods 0.000 title claims abstract description 31
- 238000007670 refining Methods 0.000 title claims abstract description 11
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000004202 carbamide Substances 0.000 claims abstract description 26
- 229920002678 cellulose Polymers 0.000 claims abstract description 6
- 239000001913 cellulose Substances 0.000 claims abstract description 5
- 240000008042 Zea mays Species 0.000 claims description 35
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 35
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 35
- 235000005822 corn Nutrition 0.000 claims description 35
- 229940088598 enzyme Drugs 0.000 claims description 31
- 239000010907 stover Substances 0.000 claims description 30
- 108010046334 Urease Proteins 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 108090000604 Hydrolases Proteins 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 6
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 5
- 239000012978 lignocellulosic material Substances 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
- 239000010902 straw Substances 0.000 claims description 4
- 239000000835 fiber Substances 0.000 claims description 3
- 244000068988 Glycine max Species 0.000 claims description 2
- 235000010469 Glycine max Nutrition 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims description 2
- 235000007164 Oryza sativa Nutrition 0.000 claims description 2
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 2
- 229920001131 Pulp (paper) Polymers 0.000 claims description 2
- 235000021307 Triticum Nutrition 0.000 claims description 2
- 244000098338 Triticum aestivum Species 0.000 claims description 2
- 239000010828 animal waste Substances 0.000 claims description 2
- 229940069765 bean extract Drugs 0.000 claims description 2
- 239000007799 cork Substances 0.000 claims description 2
- 239000011121 hardwood Substances 0.000 claims description 2
- 108010002430 hemicellulase Proteins 0.000 claims description 2
- 229940059442 hemicellulase Drugs 0.000 claims description 2
- 244000005700 microbiome Species 0.000 claims description 2
- 238000004064 recycling Methods 0.000 claims description 2
- 235000009566 rice Nutrition 0.000 claims description 2
- 230000028327 secretion Effects 0.000 claims description 2
- 238000009966 trimming Methods 0.000 claims description 2
- 210000002700 urine Anatomy 0.000 claims description 2
- 239000002699 waste material Substances 0.000 claims description 2
- 240000006394 Sorghum bicolor Species 0.000 claims 1
- 235000015505 Sorghum bicolor subsp. bicolor Nutrition 0.000 claims 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 claims 1
- 239000002023 wood Substances 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 16
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 abstract description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 8
- 229910021529 ammonia Inorganic materials 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 4
- 230000000813 microbial effect Effects 0.000 abstract description 3
- 238000000855 fermentation Methods 0.000 abstract description 2
- 230000004151 fermentation Effects 0.000 abstract description 2
- 230000010354 integration Effects 0.000 abstract description 2
- 238000007781 pre-processing Methods 0.000 abstract description 2
- 229920002488 Hemicellulose Polymers 0.000 abstract 1
- 150000003863 ammonium salts Chemical class 0.000 abstract 1
- 150000002772 monosaccharides Chemical class 0.000 abstract 1
- 229920001503 Glucan Polymers 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 11
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 7
- 239000008103 glucose Substances 0.000 description 7
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 6
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- 235000010980 cellulose Nutrition 0.000 description 4
- 229920005610 lignin Polymers 0.000 description 4
- 108010059892 Cellulase Proteins 0.000 description 3
- 229940106157 cellulase Drugs 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical group O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 239000011435 rock Substances 0.000 description 2
- 230000001360 synchronised effect Effects 0.000 description 2
- 244000062793 Sorghum vulgare Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 239000002803 fossil fuel Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 235000019713 millet Nutrition 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- -1 sawdust Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 150000003742 xyloses Chemical class 0.000 description 1
Classifications
-
- 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
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention provides a kind of lignocellulosic method for refining for synchronizing and carrying out pre-processing with enzyme hydrolysis, belong to biorefinery technical field.Lignocellulosic is directly converted to fermentable sugar using urea and hydrolase by the present invention, and urea generates gaseous ammonia or ammonium salt destroys lignocellulosic structure, cellulose and hemicellulose are converted to monosaccharide by hydrolase under the action of urase.Urea, which not only facilitates, improves enzyme hydrolysis effect, is also used as the nitrogen source of microbial fermentation.Present invention process realizes pretreatment and enzyme hydrolysis process integration, reduces the industrialized cost of lignocellulosic.
Description
Technical field
The invention belongs to biorefinery technical fields, are related to a kind of lignocellulosic for synchronizing and carrying out pretreatment and enzyme hydrolysis
Method for refining.
Background technique
Lignocellulosic is expected to substitute fossil fuels as the renewable resource being most easily obtained extensively.Currently, wooden fibre
It ties up in plain conversion process, pretreatment is the technical step of most critical.By pre-treatment step, the structure of lignocellulosic is destroyed,
The structure for changing cellulose, makes more celluloses and cellulase be in contact, and then be converted to sugar fermentation.Traditional pretreatment
Step energy consumption is high, and after pretreatment, needs to cool down or detoxification carries out next step operation, these all hinder lignocellulosic
Industrialized production.
Ammonia process pretreatment is a kind of weak base pretreatment, has hypotoxicity and low-corrosiveness, under cryogenic can be efficient
Delignification, and have many advantages, such as Ke Xunhuanliyong.However, ammonia process pretreatment exists such as equipment requirement height, high compression, ammonia time
The disadvantages of receiving unit is at high cost is unfavorable for applying on a large scale.In the pretreatment of many ammonia process, urea pretreatment has a lot excellent
Gesture, comprising: (1) inexpensive, the reagent of high nitrogen；(2) as the nitrogen source of microbial growth and metabolism；(3) urea is relative to ammonia side
Just it transports, and small (Wang L, the et al.High-solid pretreatment of corn stover using of risk
urea for enzymatic saccharification.[J].Bioresour Technol,2018,259:83-90.).And
And urea reduces lignin with hydrolysis by the electrostatic attraction between reduction hydrophobic interaction and lignin and cellulase
The absorption of enzyme improves the transformation efficiency of sugar.Hydroxyl on urea and water, lignin forms hydroxyl, and destructive enzyme is between lignin
The connection of hydrogen bond.So enzyme hydrolysis effect can be enhanced in addition urea during enzyme hydrolysis.60g/ is added during enzyme hydrolysis
The urea of L, the digestibility of substrate improve 33.1% (Lou H, et al.Effect of Urea on the Enzymatic
Hydrolysis of Lignocellulosic Substrate and Its Mechanism.Bioenergy Research,
2018,11(4):1-10.)。
Summary of the invention
For the limitation that current pretreatment and enzyme hydrolysis process separately carry out, the present invention provides a kind of synchronize and pre-processes
With the lignocellulosic method for refining of enzyme hydrolysis.This method is reduced wooden using urea progress biorefinery synchronous with hydrolase
Cellulose is converted to the cost of target product.
Technical scheme is as follows:
Synchronize the lignocellulosic method for refining for carrying out pretreatment and enzyme hydrolysis, comprising the following steps:
By after crushing or granulating lignocellulosic material, urea liquid, mix containing urease reagents and hydrolase it is equal
It is even, it is heated to 30 DEG C~60 DEG C and reacts 0.5~10 day；Or it is lignocellulosic material, the urea after crushing or granulating is molten
It liquid and is uniformly mixed containing urease reagents, is heated to 30 DEG C~60 DEG C and reacts 0.5~10 day, be then down to 60 DEG C hereinafter, water is added
Enzyme is solved, enzymatic hydrolysis reaction is carried out, is reacted 1~5 day.
In the present invention, the lignocellulosic is selected from wheat stalk, corn stover, agriculture forestry waste, rice straw, height
Fine strain of millet stalk, soybean stalk, forest trimmings, recycling wood pulp cellulose, sawdust, cork, one of hardwood and animal wastes or
It is a variety of.
In the present invention, the diameter of the crushing lignocellulosic is 0.01mm~10cm；Lignocellulosic particles
Diameter is 0.5mm~10cm.
In the present invention, the urease reagents that contain can be urase protein, bean extract, beans rhizome, animal
Urine or microorganism secretion urase albumen.
In the present invention, the hydrolase can be cellulase, hemicellulase, in pectase and zytase
It is one or more kinds of.
Compared with prior art, the invention has the following advantages that
The present invention is pre-processed using urea, and urea provides nitrogen source as high nitrogen compound, for microbial growth and metabolism,；
And urea pretreatment is synchronous with polysaccharide enzyme hydrolysis process to be carried out, and is simplified the step of pretreatment is with during enzyme hydrolysis, is saved
Process costs.
Detailed description of the invention
Fig. 1 is sugared concentration variation diagram during the Factor of Enzymolysis Corn Stalk of the UE process in embodiment 1.
Fig. 2 is the UE-CS and transformation efficiency figure sugared after untreated corn stover 13 days in embodiment 2.
Specific embodiment
To facilitate the understanding of the present invention, the present invention is done more comprehensively in conjunction with the accompanying drawings and embodiments, careful description, but this
The protection scope of invention is not limited to embodiment in detail below.
Hereafter used all technical terms are identical as the normally understood meaning of those skilled in the art, unless otherwise fixed
Justice.Technical term used herein is intended merely to describe the purpose of specific example, is not intended to and limits guarantor of the invention
Protect range.
Unless otherwise specified, the reagent used in the present invention, raw material, instrument and equipment etc. can market be commercially available
Or existing method is prepared.
Abbreviation below is used in embodiment:
" UE " is urea and enzyme hydrolysis process integration (Urea andenzyme), " CS " corn stover (Cornstover).
Embodiment 1
UE process corn stalk enzyme hydrolysis, comprising the following steps:
1, after the corn stover in farmland is collected, it raw material preparation: is ground into the particle of 1-4mm after natural drying.
2, UE technique: smashed corn stover is placed in 250mL shaking flask, and the concentration of substrate is 17.4% corn stover
(6% glucan), remaining is urea liquid, and the concentration of urea is 120g/L, and urease content is 0.1% (based on corn stover
Dry weight), be placed in 80 DEG C of baking oven, total coreaction 13 days, every other day take out and rock primary, the 7th day, hydrolysis is added
Enzyme under the conditions of 250rpm, carries out enzymatic hydrolysis reaction, the reaction time is remaining 6 days at 50 DEG C.
In the present embodiment, UE-CS enzyme hydrolysis effect curve under the conditions of 17.4% corn stover (6% glucan), such as
Shown in Fig. 1.Indicate the concentration of 17.4% corn stover (6% glucan) enzyme hydrolysis arabinose,
Indicate the concentration of 17.4% corn stover (6% glucan) enzyme hydrolysis xylose,Indicate 17.4% corn stover
The concentration of (6% glucan) enzyme hydrolysis glucose,Indicate 17.4% corn stover (6% glucan) enzyme hydrolysis
The concentration of cellobiose.Corn stover passes through UE technique as can be seen from Figure 1, and concentration of glucose can reach 49.4g/ within the 10th day
L, xylose concentration also have 12.7g/L.At subsequent three days, enzyme hydrolysis rate slowed down, and concentration of glucose reaches 57.5g/L after 13 days,
Xylose concentration is 14.0g/L, and cellobiose concentration also reaches 8.0g/L.Show the process corn stover, enzyme hydrolysis
The transformation efficiency of considerable sugar is realized in journey, for pretreatment and enzyme hydrolysis synchronize carry out providing the possibility of realization, be also fibre at present
It ties up plain ethyl alcohol production and a kind of new technique is provided.
Embodiment 2
The corn stover of UE processing and the comparison of unpretreated corn stalk sugar conversion ratio, comprising the following steps:
1, after the corn stover in farmland is collected, it raw material preparation: is ground into the particle of 1-4mm after natural drying.
2, UE technique: smashed corn stover is placed in 250mL shaking flask, and the concentration of substrate is 17.4% corn stover
(6% glucan), remaining is urea liquid, and the concentration of urea is 120g/L, and urease content is 0.1% (based on corn stover
Dry weight), be placed in 80 DEG C of baking oven, total coreaction 13 days, every other day take out and rock primary, the 7th day, hydrolysis is added
Enzyme under the conditions of 250rpm, carries out enzymatic hydrolysis reaction, the reaction time is remaining 6 days at 50 DEG C.
3, the enzyme hydrolysis of corn stover is not pre-processed: smashed corn stover is placed in the shaking flask of 250ml, substrate
Concentration is 6% glucan corn stover, and hydrolase and water is added, and enzyme digestion reaction is carried out in 50 DEG C of oscillation casees (250rpm),
Reaction 13 days.
In the present embodiment, UE-CS and unpretreated corn stover are in 17.4% corn stover (6% glucan) substrate
Under the conditions of, the comparison of enzyme hydrolysis conversion ratio, as shown in Fig. 2,Indicate 17.4% corn stover (6% glucan) enzyme
The concentration of hydrolyzation of glucose,Indicate the concentration of 17.4% corn stover (6% glucan) enzyme hydrolysis xylose.From Fig. 2
It can be seen that UE-CS technique, the conversion ratio of glucose is 82% after 13 days, and without the glucose of pretreated corn stover
Conversion ratio be 36%.UE ratio does not pre-process Factor of Enzymolysis Corn Stalk, and the conversion ratio of glucose improves 46%.UE-CS technique
The conversion ratio of 13 days xyloses is 41%, is significantly improved compared to the conversion ratio for not pre-processing Factor of Enzymolysis Corn Stalk xylose.Show
UE process advan is in the conversion ratio for the sugar for improving corn stover.
Claims (5)
1. synchronizing the lignocellulosic method for refining for carrying out pretreatment and enzyme hydrolysis, which comprises the following steps:
By after crushing or granulating lignocellulosic material, urea liquid, be uniformly mixed containing urease reagents and hydrolase, add
Heat to 30 DEG C~60 DEG C react 0.5~10 day；Or by after crushing or granulating lignocellulosic material, urea liquid and contain
Urease reagents uniformly mix, be heated to 30 DEG C~60 DEG C react 0.5~10 day, be then down to 60 DEG C hereinafter, be added hydrolase, into
Row enzymatic hydrolysis reaction reacts 1~5 day.
2. lignocellulosic method for refining according to claim 1, which is characterized in that the lignocellulosic is selected from small
Wheat Straw, corn stover, agriculture forestry waste, rice straw, broomcorn straw, soybean stalk, forest trimmings, recycling wood pulp cellulose, wood
Bits, cork, one or more of hardwood and animal wastes.
3. lignocellulosic method for refining according to claim 1, which is characterized in that the crushing lignocellulosic
Diameter is 0.01mm~10cm；The diameter of lignocellulosic particles is 0.5mm~10cm.
4. lignocellulosic method for refining according to claim 1, which is characterized in that described is selected from urea containing urease reagents
Zymoprotein, bean extract, beans rhizome, the urine of animal or the urase albumen of microorganism secretion.
5. lignocellulosic method for refining according to claim 1, which is characterized in that the hydrolase can be fiber
Plain enzyme, hemicellulase, one or more of pectase and zytase.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112844324A (en) * | 2019-11-26 | 2021-05-28 | 南京理工大学 | Lignin/manganese oxide composite adsorption material and preparation method and application thereof |
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|---|---|---|---|---|
| US20060104939A1 (en) * | 2002-08-02 | 2006-05-18 | Covington Anthony D | Degrading lignocellulosic materials |
| US20090162923A1 (en) * | 2007-12-20 | 2009-06-25 | Young Jeffrey W | Methods and Compositions for Digestion of Organic Waste |
| CN105755071A (en) * | 2016-04-01 | 2016-07-13 | 江苏大学 | Method for preparing fermentable sugar or bioethanol by utilizing biomass and 'one-pot method' |
-
2019
- 2019-03-19 CN CN201910209583.1A patent/CN109913507A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060104939A1 (en) * | 2002-08-02 | 2006-05-18 | Covington Anthony D | Degrading lignocellulosic materials |
| US20090162923A1 (en) * | 2007-12-20 | 2009-06-25 | Young Jeffrey W | Methods and Compositions for Digestion of Organic Waste |
| CN105755071A (en) * | 2016-04-01 | 2016-07-13 | 江苏大学 | Method for preparing fermentable sugar or bioethanol by utilizing biomass and 'one-pot method' |
Non-Patent Citations (4)
| Title |
|---|
| HONGMING LOU等: "Effect of Urea on the Enzymatic Hydrolysis of Lignocellulosic Substrate and Its Mechanism", 《BIOENERGY RESEARCH》 * |
| WANG L等: "High-solid pretreatment of corn stover using urea for enzymatic saccharification", 《BIORESOUR TECHNOL》 * |
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| 魏蔚等: "复合菌剂对玉米秸秆的降解及土壤生态特性的影响", 《土壤通报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112844324A (en) * | 2019-11-26 | 2021-05-28 | 南京理工大学 | Lignin/manganese oxide composite adsorption material and preparation method and application thereof |
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Application publication date: 20190621 |