CN113528586A - 以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺 - Google Patents
以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺 Download PDFInfo
- Publication number
- CN113528586A CN113528586A CN202010289312.4A CN202010289312A CN113528586A CN 113528586 A CN113528586 A CN 113528586A CN 202010289312 A CN202010289312 A CN 202010289312A CN 113528586 A CN113528586 A CN 113528586A
- Authority
- CN
- China
- Prior art keywords
- lignin
- pretreatment
- ethanol
- polyhydroxyalkanoate
- lignocellulose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 124
- 239000005014 poly(hydroxyalkanoate) Substances 0.000 title claims abstract description 53
- 229920000903 polyhydroxyalkanoate Polymers 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 40
- 239000002994 raw material Substances 0.000 title claims abstract description 21
- 229920005610 lignin Polymers 0.000 claims abstract description 61
- 238000000855 fermentation Methods 0.000 claims abstract description 27
- 230000004151 fermentation Effects 0.000 claims abstract description 27
- 239000001913 cellulose Substances 0.000 claims abstract description 13
- 229920002678 cellulose Polymers 0.000 claims abstract description 13
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- 241000589776 Pseudomonas putida Species 0.000 claims abstract description 11
- 239000002699 waste material Substances 0.000 claims abstract description 11
- 229920002488 Hemicellulose Polymers 0.000 claims abstract description 9
- 229940059442 hemicellulase Drugs 0.000 claims abstract description 7
- 108010002430 hemicellulase Proteins 0.000 claims abstract description 7
- 239000010902 straw Substances 0.000 claims description 15
- 108090000790 Enzymes Proteins 0.000 claims description 9
- 102000004190 Enzymes Human genes 0.000 claims description 9
- 229940088598 enzyme Drugs 0.000 claims description 9
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 claims description 8
- 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 claims description 8
- 238000003776 cleavage reaction Methods 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 230000007017 scission Effects 0.000 claims description 8
- 240000008042 Zea mays Species 0.000 claims description 7
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 7
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 7
- 235000005822 corn Nutrition 0.000 claims description 7
- 108010059892 Cellulase Proteins 0.000 claims description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 6
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 6
- 229940106157 cellulase Drugs 0.000 claims description 6
- 230000002255 enzymatic effect Effects 0.000 claims description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 4
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims description 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims description 4
- 239000000413 hydrolysate Substances 0.000 claims description 4
- 239000002351 wastewater Substances 0.000 claims description 4
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 3
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 3
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 239000010907 stover Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 244000068988 Glycine max Species 0.000 claims description 2
- 235000010469 Glycine max Nutrition 0.000 claims description 2
- 108010029541 Laccase Proteins 0.000 claims description 2
- 241001465754 Metazoa Species 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims description 2
- 235000007164 Oryza sativa Nutrition 0.000 claims description 2
- 102000003992 Peroxidases Human genes 0.000 claims description 2
- 108010059820 Polygalacturonase Proteins 0.000 claims description 2
- 229920001131 Pulp (paper) Polymers 0.000 claims description 2
- 240000006394 Sorghum bicolor Species 0.000 claims description 2
- 235000011684 Sorghum saccharatum Nutrition 0.000 claims description 2
- 235000021307 Triticum Nutrition 0.000 claims description 2
- 244000098338 Triticum aestivum Species 0.000 claims description 2
- 239000003513 alkali Substances 0.000 claims description 2
- 229910021529 ammonia Inorganic materials 0.000 claims description 2
- 238000004517 catalytic hydrocracking Methods 0.000 claims description 2
- 108010093305 exopolygalacturonase Proteins 0.000 claims description 2
- 238000004880 explosion Methods 0.000 claims description 2
- 210000003608 fece Anatomy 0.000 claims description 2
- 239000000835 fiber Substances 0.000 claims description 2
- 239000011121 hardwood Substances 0.000 claims description 2
- 239000002608 ionic liquid Substances 0.000 claims description 2
- 239000010871 livestock manure Substances 0.000 claims description 2
- 239000003960 organic solvent Substances 0.000 claims description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 2
- 238000002203 pretreatment Methods 0.000 claims description 2
- 235000009566 rice Nutrition 0.000 claims description 2
- 239000011122 softwood Substances 0.000 claims description 2
- 239000002023 wood Substances 0.000 claims description 2
- 230000003197 catalytic effect Effects 0.000 claims 1
- 238000007670 refining Methods 0.000 abstract description 12
- 239000007788 liquid Substances 0.000 abstract description 10
- 238000005336 cracking Methods 0.000 abstract 1
- 230000003301 hydrolyzing effect Effects 0.000 abstract 1
- 238000000197 pyrolysis Methods 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 description 12
- 229910052799 carbon Inorganic materials 0.000 description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 230000007062 hydrolysis Effects 0.000 description 5
- 238000006460 hydrolysis reaction Methods 0.000 description 5
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- 241001052560 Thallis Species 0.000 description 4
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- -1 fatty acid ester Chemical class 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 238000004566 IR spectroscopy Methods 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000035508 accumulation Effects 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 150000001721 carbon Chemical group 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000002026 chloroform extract Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 150000001491 aromatic compounds Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006065 biodegradation reaction Methods 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000006652 catabolic pathway Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 230000007154 intracellular accumulation Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 239000010841 municipal wastewater Substances 0.000 description 1
- VOFUROIFQGPCGE-UHFFFAOYSA-N nile red Chemical compound C1=CC=C2C3=NC4=CC=C(N(CC)CC)C=C4OC3=CC(=O)C2=C1 VOFUROIFQGPCGE-UHFFFAOYSA-N 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920006255 plastic film Polymers 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000002910 solid waste Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000004065 wastewater treatment Methods 0.000 description 1
Images
Classifications
-
- 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
-
- 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
- C12P7/62—Carboxylic acid esters
- C12P7/625—Polyesters of hydroxy carboxylic acids
-
- 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
-
- 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
- C12P2203/00—Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
-
- 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
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明公开了一种以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺。所述工艺先将木质纤维素原料预处理得到纤维素、半纤维素和木质素，然后将纤维素和半纤维素酶水解，再以酶解液为培养基，接种乙醇发酵菌株生产乙醇，然后直接以预处理后得到的木质素和乙醇发酵产生的含木质素残渣为培养基，或将预处理后得到的木质素和乙醇发酵产生的含木质素残渣经过裂解后得到的裂解液为培养基，接种恶臭假单胞菌Pseudomonas putida NX‑1生产聚羟基脂肪酸酯。本发明将以木质素为原料生产聚羟基脂肪酸酯的工艺和纤维素生产乙醇工艺进行集成，实现木质纤维素三种主要组分的充分利用，同时降低了炼制过程中废弃物的产生，提高整个木质纤维素炼制过程的经济性。
Description
技术领域
本发明属于生物炼制技术领域，涉及一种以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺。
背景技术
木质纤维素是地球上最丰富的可再生资源，其主要成分为纤维素、半纤维素和木质素。随着预处理、酶水解及发酵菌株开发等技术的不断进步，木质纤维素生物炼制水平得到了较大提升。目前生物乙醇是木质纤维素炼制的代表性产品，世界上许多国家近些年都在大力推进木质纤维素炼制生物乙醇的技术研发及产业化(Nature ReviewsMicrobiology,2016,14(5):288；中国科技成果,2015,21:38-39)。但几乎所有木质纤维素生物炼制过程都会产生大量富含木质素的废液或固体废弃物。
木质素是自然界中最丰富的芳香族化合物，不同物种来源的木质纤维素中木质素含量可达其总质量的10％-35％。随着木质纤维素炼制乙醇等大宗能源及化学品技术的快速发展，木质纤维素中的纤维素和半纤维素被转化为燃料乙醇或化学品，木质素则被作为残渣剩余。根据原料不同，生产一升纤维素乙醇将伴随0.5-1.5千克木质素残渣的生成(Green Chemistry,2015,17(11):4860-4861)。在当前木质纤维素生物炼制理念中，生产的木质素多被用来焚烧供热、供能，同时储存在木质素中的碳原子以二氧化碳的形式排放到大气中，这种粗暴的焚烧供能策略在一定意义上会造成“碳原子流失”。将木质素转化为附加值更高的化学品对其价值提升及资源利用均具有重要意义。
聚羟基脂肪酸酯是可生物降解且具有生物相容性的天然聚酯，许多微生物在营养限制但存在过量碳源的情况下将其作为碳源和能量存储在细胞内。由于其通用的材料特性，聚羟基脂肪酸酯被认为是石油化学衍生塑料的潜在替代品之一。虽然聚羟基脂肪酸酯的微生物合成、规模化生产与在不同行业中的应用均得到了快速发展，但微生物生产聚羟基脂肪酸酯的工业化仍面临生产成本过高的问题，其中原料成本是聚羟基脂肪酸酯生产成本的重要组成部分。目前最主要的聚羟基脂肪酸酯生产原料为淀粉、葡萄糖及甘油(International Biodeterioration&Biodegradation,2018,126:45-56)。为降低聚羟基脂肪酸酯的原料成本，许多研究人员也尝试使用廉价原料来生产聚羟基脂肪酸酯，包括市政废水、造纸废水、活性污泥、食品生产废弃物等。近年来，研究人员发现一些木质素降解菌自身除了含有丰富的木质素解聚和降解途径，还含有一些产物合成途径，其中就包括聚羟基脂肪酸酯的合成途径(Renewable and Sustainable Energy Reviews,2017,73:610-621)。完备的木质素降解-聚羟基脂肪酸酯合成途径使得以木质素为原料生产聚羟基脂肪酸酯成为了可能。
发明内容
针对木质纤维素炼制生产乙醇的过程中产生的大量木质素无法利用的问题，本发明提供一种以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺。该工艺将以木质素为原料生产聚羟基脂肪酸酯的工艺和纤维素生产乙醇的工艺进行集成，分别将“纤维素+半纤维素组分和木质素组分转化为生物乙醇和聚羟基脂肪酸酯，实现木质纤维素三种主要组分的充分利用。
本发明的技术方案如下：
以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺，包括以下步骤：
步骤1，将木质纤维素原料进行预处理得到纤维素、半纤维素和木质素；
步骤2，将纤维素和半纤维素用纤维素酶和半纤维素酶进行酶水解，得到酶解液；
步骤3，以酶解液为培养基，接种乙醇发酵菌株，发酵生产乙醇；
步骤4，直接以预处理后得到的木质素和步骤3中乙醇发酵产生的含木质素残渣为培养基，或将预处理后得到的木质素和步骤3中乙醇发酵产生的含木质素残渣经过裂解后得到的裂解液为培养基，接种恶臭假单胞菌Pseudomonas putida NX-1，发酵生产聚羟基脂肪酸酯。
步骤1中，所述的木质纤维素原料为本领域常规使用的木质纤维素原料，可以是小麦秸秆、玉米秸秆、农林废料、水稻秸秆、高粱秸秆、大豆秸秆、林业废料、回收木浆纤维、木屑、软木、硬木或动物粪便等。
步骤1中，所述的预处理的方法为本领域常规使用的方法，可以是酸预处理、碱预处理、热水预处理、蒸汽爆破预处理、氨预处理、有机溶剂预处理、离子液预处理或两种以上的联合预处理。
步骤2中，所述的纤维素酶和半纤维素酶为常规使用的纤维素酶和半纤维素酶，例如果胶酶、木聚糖酶等。
步骤3中，所述的乙醇发酵菌株为常规使用的乙醇发酵菌株，可以为单独利用葡萄糖的菌株，单独利用木糖的菌株，以及同时利用葡萄糖和木糖的菌株，例如酿酒酵母Saccharomyces cerevisiae ATCC 26603等。
步骤4中，所述的预处理后得到的木质素可以是预处理后分离得到的木质素固体和/或溶解在废水中的木质素。
步骤4中，所述的裂解的方法为本领域常规使用的方法，可以是高温高压裂解、加氢裂解、使用催化剂裂解或者使用漆酶、过氧化物酶等生物酶进行裂解。
与现有技术相比，本发明具有以下优点：
(1)利用木质纤维素联合生产乙醇和聚羟基脂肪酸酯，利用纤维素和半纤维素组分生产乙醇，木质素组分生产聚羟基脂肪酸酯，不仅实现了木质纤维素三种主要组分的全利用，而且相比传统木质纤维素炼制过程仅获得单一乙醇产品外，还获得了聚羟基脂肪酸酯，一定程度地提高了木质纤维素炼制的碳原子经济性。
(2)现有木质素通常是用来焚烧功能，焚烧过程中储存在木质素的碳原子转变成本CO2释放入大气中，会加剧温室效应的产生。将木质素转变为聚羟基脂肪酸酯则可将碳原子固定在聚羟基脂肪酸酯这一生物材料中，而不是CO2释放入大气中，由此更加有利于环境保护。
(3)木质纤维素炼制废水中通常会含有大量木质素相关的物质，造成后期废水处理困难，本工艺在将废水中的木质素转化为聚羟基脂肪酸酯的同时，也降低了废水中木质素的含量，降低了后续废水处理的成本。
综上所述的，本发明工艺在实现木质纤维素原料到乙醇和聚羟基脂肪酸酯生产的同时，降低了炼制过程中废弃物的产生，有助于提高整个木质纤维素炼制过程的经济性。
附图说明
图1为以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺流程示意图，本工艺中可以用来进行聚羟基脂肪酸酯生产的木质素有四种来源：①预处理之后含木质素的废液；②预处理之后萃取得到的木质素固体；③酶水解后含木质素的残渣；④乙醇发酵结束后含木质素的残渣。
图2为酿酒酵母Saccharomyces cerevisiae ATCC 26603利用木质纤维素原料中的纤维素组分发酵生产乙醇的发酵图。
图3为恶臭假单胞菌Pseudomonas putida NX-1利用木质素发酵生产聚羟基脂肪酸酯的发酵图。
图4为用荧光法检测以木质素为碳源培养的臭假单胞菌Pseudomonas putida NX-1胞内累计的聚羟基脂肪酸酯(A)；及将恶臭假单胞菌Pseudomonas putida NX-1胞内的聚羟基脂肪酸酯提取后制成的膜。
图5为利用红外光谱检测、核磁共振等检测手段对以木质素为碳源培养的臭假单胞菌Pseudomonas putida NX-1胞内累计的聚羟基脂肪酸酯进行结构鉴定，A：红外光谱检测图谱；B：核磁共振检测氢谱图；C：核磁共振碳谱图；D：根据上述检测推测的聚羟基脂肪酸酯结构图。
具体实施方式
下面结合实施例和附图对本发明作进一步详述。
下述实施例中，采用的酿酒酵母Saccharomyces cerevisiae ATCC 26603为商业购买的菌株，恶臭假单胞菌Pseudomonas putida NX-1为发明人实验室筛选得到的菌株，已在文献(Environmental Science and Pollution Research,2018,25(2):1-11)公开，并已在中国普通微生物菌种保藏管理中心进行保藏，保藏编号为CGMCC15317。
实施例1
1.通过稀酸预处理玉米秸秆
将玉米秸秆按10％(w/w)的干物浓度加入1％(w/w)硫酸溶液中，用高压灭菌锅在160℃处理10min，随后用真空抽滤泵将液体抽出备用。
2.预处理后的玉米秸秆经过酶水解后发酵产乙醇
预处理后的玉米秸秆按30％(w/w)底物浓度进行酶水解，总酶的添加量为40mg酶/g葡聚糖，纤维素酶与木聚糖酶的比例为8：2，在50℃、250rpm条件下酶解72h，之后过滤得到酶解液。
以上述酶解液作为培养基，将酿酒酵母Saccharomyces cerevisiae ATCC 26603以初始OD600＝1的量接种至酶解液中，厌氧条件下30℃、250rpm培养144h。分别在4h、14h、24h、36h、48h、60h、72h、84h、96h、108h、120h、132h、144h取样，离心后上清液用高效液相色谱检测葡萄糖的消耗量及乙醇的产量。绘制菌体生长、产物合成及底物消耗曲线。
3.利用木质素生产聚羟基脂肪酸酯
将酶水解后含木质素的残渣为原料(图1木质素来源③获得)，以10g/L的添加量加入到M9培养基(Na2HPO4·12H2O 6.78g/L；KH2PO4 3g/L；NaCl 0.5g/L；NH4Cl 0.05g/L；MgSO4·7H2O 0.246g/L；CaCl2 0.056g/L；Hoagland solution溶液10mL/L)中，配置成含有木质素的M9培养基，使用HCl调节pH为7.0。将培养好的恶臭假单胞菌Pseudomonas putidaNX-1种子细胞以初始OD600＝0.2接入600mL含有木质素的基础盐培养基中，250rpm、30℃培养144h，期间每隔24h取样一次。通过尼罗红将细胞染色，在激光共聚焦显微镜下观察胞内聚羟基脂肪酸酯的积累情况。菌体冻干后使用氯仿法提取聚羟基脂肪酸酯，并计算聚羟基脂肪酸酯含量和浓度。使用FT-IR、NMR等检测确定聚羟基脂肪酸酯的结构。
氯仿法提取聚羟基脂肪酸酯的方法如下：将获得的发酵液样品离心弃上清，使用去离子水洗涤两遍并收集菌体，将获得的菌体真空冷冻干燥后，按1g干细胞/10mL氯仿的比例加入抽提瓶中，100℃密封加热抽提4h。冷却后，使用0.22μm有机滤膜过滤掉细胞碎片，将澄清的氯仿抽提液加热蒸发至1/5体积，加入10倍体积的乙醇沉淀聚羟基脂肪酸酯。为使得聚羟基脂肪酸酯充分沉淀，将含有乙醇的氯仿抽提液置于4℃环境中沉淀24h。
图1为以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺流程示意图。图中玉米秸秆经过不同预处理方式使木质纤维素结构破坏，经预处理后的秸秆再经过酶解(纤维素酶、半纤维素酶)进一步发酵产乙醇，而此过程中产生的含木质素的废弃物(液)可用来发酵产聚羟基脂肪酸酯。如图所示，其中可以用来生产聚羟基脂肪酸酯的木质素主要来源于：秸秆预处理后产生的含木质素废液和不溶性木质素(图1中①、②)；将预处理后的秸秆进行酶水解，之后固液分离得到的木质素残渣(图1中③)；预处理后的秸秆同步糖化发酵产乙醇，发酵结束后的含有木质素的残渣(图1中④)。
图2为酿酒酵母Saccharomyces cerevisiae ATCC 26603利用木质纤维素原料中的纤维素组分发酵生产乙醇的发酵图，图中展示了菌体生长、所产乙醇浓度、葡萄糖浓度分别与时间的关系曲线。发酵144h后，葡萄糖消耗完毕，生产乙醇38.89g/L。
图3为恶臭假单胞菌Pseudomonas putida NX-1利用木质素发酵生产聚羟基脂肪酸酯的发酵图。
图4中，A为激光共聚焦显微镜观察到的木质素培养的Pseudomonas putida NX-1细胞内聚羟基脂肪酸酯的积累情况；B为将聚羟基脂肪酸酯提取后制成的塑料膜。
图5中，A为聚羟基脂肪酸酯的红外光谱图，存在聚羟基脂肪酸酯的典型官能团-C＝O；B、C为所产聚羟基脂肪酸酯的氢谱、碳谱图；D为所产聚羟基脂肪酸酯结构式，说明得到的聚羟基脂肪酸脂含有四种单体，碳链长度分别为8个、10个、12个、14个碳原子。
Claims (8)
1.以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺，其特征在于，包括以下步骤：
步骤1，将木质纤维素原料进行预处理得到纤维素、半纤维素和木质素；
步骤2，将纤维素和半纤维素用纤维素酶和半纤维素酶进行酶水解，得到酶解液；
步骤3，以酶解液为培养基，接种乙醇发酵菌株，发酵生产乙醇；
步骤4，直接以预处理后得到的木质素和步骤3中乙醇发酵产生的含木质素残渣为培养基，或将预处理后得到的木质素和步骤3中乙醇发酵产生的含木质素残渣经过裂解后得到的裂解液为培养基，接种恶臭假单胞菌Pseudomonas putida NX-1，发酵生产聚羟基脂肪酸酯。
2.根据权利要求1所述的工艺，其特征在于，步骤1中，所述的木质纤维素原料选自小麦秸秆、玉米秸秆、农林废料、水稻秸秆、高粱秸秆、大豆秸秆、林业废料、回收木浆纤维、木屑、软木、硬木或动物粪便。
3.根据权利要求1所述的工艺，其特征在于，步骤1中，所述的预处理的方法选自酸预处理、碱预处理、热水预处理、蒸汽爆破预处理、氨预处理、有机溶剂预处理、离子液预处理或两种以上的联合预处理。
4.根据权利要求1所述的工艺，其特征在于，步骤2中，所述的半纤维素酶为果胶酶或木聚糖酶。
5.根据权利要求1所述的工艺，其特征在于，步骤3中，所述的乙醇发酵菌株为单独利用葡萄糖的菌株，单独利用木糖的菌株，或同时利用葡萄糖和木糖的菌株。
6.根据权利要求1所述的工艺，其特征在于，步骤3中，所述的乙醇发酵菌株为酿酒酵母Saccharomyces cerevisiae ATCC 26603。
7.根据权利要求1所述的工艺，其特征在于，步骤4中，所述的预处理后得到的木质素为预处理后分离得到的木质素固体和/或溶解在废水中的木质素。
8.根据权利要求1所述的工艺，其特征在于，步骤4中，所述的裂解的方法为高温高压裂解、加氢裂解、使用催化剂裂解或者使用漆酶或过氧化物酶的生物酶进行裂解。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010289312.4A CN113528586A (zh) | 2020-04-14 | 2020-04-14 | 以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010289312.4A CN113528586A (zh) | 2020-04-14 | 2020-04-14 | 以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113528586A true CN113528586A (zh) | 2021-10-22 |
Family
ID=78120050
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010289312.4A Pending CN113528586A (zh) | 2020-04-14 | 2020-04-14 | 以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113528586A (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114277067A (zh) * | 2021-12-27 | 2022-04-05 | 常州柯纳生物科技有限公司 | 一种利用韧皮纤维生产pha材料的方法 |
| CN114294905A (zh) * | 2021-11-19 | 2022-04-08 | 中粮生物科技股份有限公司 | 利用红外或微波干燥聚羟基脂肪酸酯的方法 |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586339A (zh) * | 2012-02-20 | 2012-07-18 | 清华大学 | 一种甜高粱秆联产燃料乙醇和木质素的方法 |
| CN104726502A (zh) * | 2015-04-03 | 2015-06-24 | 南京林业大学 | 一种纤维素废弃物生物制备乙醇并联产壳聚糖的方法 |
| CN105400830A (zh) * | 2015-12-29 | 2016-03-16 | 四川大学 | 一种利用木质纤维素原料生产乙醇的方法 |
| KR20160068391A (ko) * | 2014-12-05 | 2016-06-15 | 서울대학교산학협력단 | 생분해성 폴리하이드록시알카노에이트 및 이의 제조방법 |
| CN108949842A (zh) * | 2018-07-20 | 2018-12-07 | 中南大学 | 一种木质纤维素酸法预处理废液的资源化利用方法 |
| US20180371502A1 (en) * | 2013-12-06 | 2018-12-27 | Alliance For Sustainable Energy, Llc | Lignin conversion to fuels, chemicals and materials |
-
2020
- 2020-04-14 CN CN202010289312.4A patent/CN113528586A/zh active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586339A (zh) * | 2012-02-20 | 2012-07-18 | 清华大学 | 一种甜高粱秆联产燃料乙醇和木质素的方法 |
| US20180371502A1 (en) * | 2013-12-06 | 2018-12-27 | Alliance For Sustainable Energy, Llc | Lignin conversion to fuels, chemicals and materials |
| KR20160068391A (ko) * | 2014-12-05 | 2016-06-15 | 서울대학교산학협력단 | 생분해성 폴리하이드록시알카노에이트 및 이의 제조방법 |
| CN104726502A (zh) * | 2015-04-03 | 2015-06-24 | 南京林业大学 | 一种纤维素废弃物生物制备乙醇并联产壳聚糖的方法 |
| CN105400830A (zh) * | 2015-12-29 | 2016-03-16 | 四川大学 | 一种利用木质纤维素原料生产乙醇的方法 |
| CN108949842A (zh) * | 2018-07-20 | 2018-12-07 | 中南大学 | 一种木质纤维素酸法预处理废液的资源化利用方法 |
Non-Patent Citations (1)
| Title |
|---|
| 宋存江等: "《制浆造纸产业生物质精炼发展研究》", 中国轻工业出版社, pages: 364 - 25 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114294905A (zh) * | 2021-11-19 | 2022-04-08 | 中粮生物科技股份有限公司 | 利用红外或微波干燥聚羟基脂肪酸酯的方法 |
| CN114277067A (zh) * | 2021-12-27 | 2022-04-05 | 常州柯纳生物科技有限公司 | 一种利用韧皮纤维生产pha材料的方法 |
| WO2023125546A1 (zh) * | 2021-12-27 | 2023-07-06 | 常州柯纳生物科技有限公司 | 一种利用韧皮纤维生产pha材料的方法 |
| CN114277067B (zh) * | 2021-12-27 | 2023-08-04 | 常州柯纳生物科技有限公司 | 一种利用韧皮纤维生产pha材料的方法 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Selvakumar et al. | Optimization of binary acids pretreatment of corncob biomass for enhanced recovery of cellulose to produce bioethanol | |
| CN101514349B (zh) | 一种由竹材纤维制备燃料乙醇的方法 | |
| US20140227757A1 (en) | Integrated processes for conversion of lignocellulosic biomass to bioproducts and systems and apparatus related thereto | |
| CN102154381B (zh) | 一种以木质纤维素为原料联产乙醇和微生物油脂的方法 | |
| Jafari et al. | Efficient bioconversion of whole sweet sorghum plant to acetone, butanol, and ethanol improved by acetone delignification | |
| Junluthin et al. | Efficient conversion of night-blooming giant water lily into bioethanol and biogas | |
| WO2013106113A2 (en) | Integrated processes for conversion of lignocellulosic biomass to bioproducts and systems and apparatus related thereto | |
| Mohit et al. | Production of bio-ethanol from Jatropha oilseed cakes via dilute acid hydrolysis and fermentation by Saccharomyces cerevisiae | |
| CN113528586A (zh) | 以木质纤维素为原料联合生产乙醇和聚羟基脂肪酸酯的工艺 | |
| US20140242656A1 (en) | Method for producing alcohols and/or solvents from paper pulps with recycling of the non-hydrolysated plant material in a regeneration reactor | |
| KR100965851B1 (ko) | 팝핑법을 이용한 리그노셀룰로스계 바이오매스의 전처리방법, 및 이를 이용한 당화합물 및 바이오에탄올의생산방법 | |
| Eshtiaghi et al. | Preliminary study for bioconversion of water hyacinth (Eichhornia crassipes) to bioethanol | |
| Manmai et al. | Transformation of lignocellulose from corn stove for bioethanol production | |
| CN106929547A (zh) | 一种利用稻草高效联产甲烷和乙醇的方法 | |
| KR101504197B1 (ko) | 목질계 바이오매스로부터 바이오에탄올을 제조하는 방법 | |
| Roy et al. | Bioethanol Production from Lignocellulose Agricultural Waste Biomass | |
| Ylitervo | Production of ethanol and biomass from orange peel waste by Mucor indicus | |
| CN104531806A (zh) | 一种生产可发酵性糖的方法 | |
| CN102476128B (zh) | 一种城乡生活垃圾绿色转化可再生能源的方法 | |
| Gunawan et al. | Bioethanol production from rice straw waste by enzyme catalyst using separated hydrolysis and fermentation (SHF) method | |
| Taha et al. | Successive application of physicochemical and enzymatic treatments of office paper waste for the production of bioethanol with possible using of carbon dioxide as an indicator for the determination of the bioethanol concentration | |
| Kirti et al. | Bioethanol production from agro waste–Pigeon pea (Cajanus cajan (L.) Millsp.) stalk using solid state fermentation | |
| CN110894513B (zh) | 一种木质纤维素生物质联产乙醇和富含酚类生物油的方法 | |
| Aderibigbe et al. | Optimized Production of Bioethanol by Fermentation of Acid Hydrolyzed-Corn Stover Employing Saccharomyces Cerevisiae Yeast Strain | |
| CN102154128A (zh) | 一株能直接利用木糖产生酒精的异常毕赤酵母菌菌株 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211022 |