CN1570123A - 微生物两段发酵法由甘油生产1,3-丙二醇和2,3丁二醇 - Google Patents
微生物两段发酵法由甘油生产1,3-丙二醇和2,3丁二醇 Download PDFInfo
- Publication number
- CN1570123A CN1570123A CN 200410037692 CN200410037692A CN1570123A CN 1570123 A CN1570123 A CN 1570123A CN 200410037692 CN200410037692 CN 200410037692 CN 200410037692 A CN200410037692 A CN 200410037692A CN 1570123 A CN1570123 A CN 1570123A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- glycerine
- glycerol
- ammediol
- butyleneglycol
- 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.)
- Granted
Links
- 238000000855 fermentation Methods 0.000 title claims abstract description 61
- 230000004151 fermentation Effects 0.000 title claims abstract description 61
- 238000000034 method Methods 0.000 title claims abstract description 36
- OWBTYPJTUOEWEK-UHFFFAOYSA-N butane-2,3-diol Chemical compound CC(O)C(C)O OWBTYPJTUOEWEK-UHFFFAOYSA-N 0.000 title claims abstract description 19
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 14
- 244000005700 microbiome Species 0.000 title claims description 9
- YPFDHNVEDLHUCE-UHFFFAOYSA-N propane-1,3-diol Chemical compound OCCCO YPFDHNVEDLHUCE-UHFFFAOYSA-N 0.000 title abstract description 11
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 136
- 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 abstract description 19
- 239000008103 glucose Substances 0.000 claims abstract description 19
- 235000011187 glycerol Nutrition 0.000 claims description 52
- UXFQFBNBSPQBJW-UHFFFAOYSA-N 2-amino-2-methylpropane-1,3-diol Chemical compound OCC(N)(C)CO UXFQFBNBSPQBJW-UHFFFAOYSA-N 0.000 claims description 36
- 230000008569 process Effects 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 238000002156 mixing Methods 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 7
- 230000005587 bubbling Effects 0.000 claims description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 4
- 235000011089 carbon dioxide Nutrition 0.000 claims description 4
- 239000001301 oxygen Substances 0.000 claims description 4
- 229910052760 oxygen Inorganic materials 0.000 claims description 4
- 238000010612 desalination reaction Methods 0.000 claims description 3
- 229910001873 dinitrogen Inorganic materials 0.000 claims description 3
- 238000004821 distillation Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 abstract description 13
- 239000002994 raw material Substances 0.000 abstract description 6
- 238000005516 engineering process Methods 0.000 abstract 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 abstract 1
- 238000010364 biochemical engineering Methods 0.000 abstract 1
- 238000011033 desalting Methods 0.000 abstract 1
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 241000193830 Bacillus <bacterium> Species 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 229920002215 polytrimethylene terephthalate Polymers 0.000 description 7
- 238000011218 seed culture Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 230000001580 bacterial effect Effects 0.000 description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 229920000139 polyethylene terephthalate Polymers 0.000 description 4
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 241000588747 Klebsiella pneumoniae Species 0.000 description 3
- 238000010564 aerobic fermentation Methods 0.000 description 3
- 150000002009 diols Chemical class 0.000 description 3
- 238000012262 fermentative production Methods 0.000 description 3
- 239000004310 lactic acid Substances 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- -1 polytrimethylene terephthalate Polymers 0.000 description 3
- 238000007639 printing Methods 0.000 description 3
- HGINCPLSRVDWNT-UHFFFAOYSA-N Acrolein Chemical compound C=CC=O HGINCPLSRVDWNT-UHFFFAOYSA-N 0.000 description 2
- KAKZBPTYRLMSJV-UHFFFAOYSA-N Butadiene Chemical compound C=CC=C KAKZBPTYRLMSJV-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 2
- KKEYFWRCBNTPAC-UHFFFAOYSA-N Terephthalic acid Chemical compound OC(=O)C1=CC=C(C(O)=O)C=C1 KKEYFWRCBNTPAC-UHFFFAOYSA-N 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- RXKJFZQQPQGTFL-UHFFFAOYSA-N dihydroxyacetone Chemical compound OCC(=O)CO RXKJFZQQPQGTFL-UHFFFAOYSA-N 0.000 description 2
- 239000002360 explosive Substances 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- TVMXDCGIABBOFY-UHFFFAOYSA-N octane Chemical compound CCCCCCCC TVMXDCGIABBOFY-UHFFFAOYSA-N 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- DNIAPMSPPWPWGF-VKHMYHEASA-N (+)-propylene glycol Chemical compound C[C@H](O)CO DNIAPMSPPWPWGF-VKHMYHEASA-N 0.000 description 1
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 description 1
- VVSAAKSQXNXBML-UHFFFAOYSA-N 3-acetyloxybutan-2-yl acetate Chemical class CC(=O)OC(C)C(C)OC(C)=O VVSAAKSQXNXBML-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 229910021591 Copper(I) chloride Inorganic materials 0.000 description 1
- 238000005698 Diels-Alder reaction Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 1
- 241000588748 Klebsiella Species 0.000 description 1
- 241000588915 Klebsiella aerogenes Species 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 239000004902 Softening Agent Substances 0.000 description 1
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- WFCLHMCZUVCIHV-UHFFFAOYSA-N butane-2,3-diol Chemical compound CC(O)C(C)O.CC(O)C(C)O WFCLHMCZUVCIHV-UHFFFAOYSA-N 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- OXBLHERUFWYNTN-UHFFFAOYSA-M copper(I) chloride Chemical compound [Cu]Cl OXBLHERUFWYNTN-UHFFFAOYSA-M 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 239000013530 defoamer Substances 0.000 description 1
- 229940120503 dihydroxyacetone Drugs 0.000 description 1
- 238000007323 disproportionation reaction Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 229920006351 engineering plastic Polymers 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 239000002316 fumigant Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- 238000005984 hydrogenation reaction Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000003317 industrial substance Substances 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000012092 media component Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000004745 nonwoven fabric Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 230000009965 odorless effect Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 238000003068 pathway analysis Methods 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 229920000166 polytrimethylene carbonate Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- QQONPFPTGQHPMA-UHFFFAOYSA-N propylene Natural products CC=C QQONPFPTGQHPMA-UHFFFAOYSA-N 0.000 description 1
- 125000004805 propylene group Chemical group [H]C([H])([H])C([H])([*:1])C([H])([H])[*:2] 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明公开了属于生物化工技术领域,是采用前期厌氧后期好氧两段发酵的新工艺利用微生物由甘油同时生产1,3-丙二醇和2,3-丁二醇的方法,发酵过程中先将种子液加入含工业甘油或甘油发酵液的初始发酵培养基中,发酵开始3~5小时后流加甘油和葡萄糖混合溶液。通过两段发酵过程中通气量的调节以及发酵温度的控制来调控1,3-丙二醇和2,3-丁二醇的产物浓度,发酵结束时通过脱盐、蒸馏、真空精馏等步骤将1,3-丙二醇和2,3-丁二醇分离、提纯,制备产品。该方法可以有效提高1,3-丙二醇的发酵浓度、得率和生产强度,同时还可获得较高浓度的2,3-丁二醇,从而提高了的原料利用率、降低了生产成本。
Description
技术领域
本发明属于生物化工技术领域,特别涉及一种微生物两段发酵法由甘油生产1,3-丙二醇和2,3-丁二醇。
背景技术
1,3-丙二醇(PDO)是一种重要的化工原料,可作为有机溶剂应用于油墨、印染、涂料、润滑剂、抗冻剂等行业。1,3-丙二醇最主要的用途是作为聚酯和聚氨酯合成的单体,特别是与对苯二甲酸聚合生成的聚对苯二甲酸丙二酯(PTT),显示了比以1,2-丙二醇、丁二醇、乙二醇为单体合成的聚合物更优良的性能。目前全球每年消费数千万吨聚对苯二甲酸乙二酯(PET),而PTT的化学稳定性、生物可降解性等与PET相当,但耐污染性、韧性和回弹性及抗紫外性能等更优越。此外PTT纤维还具有耐磨、吸水性低、低静电等优点,可在地毯领域与尼龙竞争。它还可用于具有优良性能的无纺布、工程塑料、服装、家庭装饰、垫衬料、织物等方面。PTT被评为美国98年六大石化新产品之一,被认为将是PET的升级产品。
PTT的优越性能及市场潜力早在50年前就被人们所认识,只因原料1,3-丙二醇生产技术难度大、成本高而导致PTT很难大规模工业化生产,迄今为止,只有Dupont和Shell两家跨国公司采用传统的化学合成路线,以环氧乙烷或丙烯为原料生产仅供它们合成PTT自用的1,3-丙二醇。化学合成法的缺点是副产物多,选择性差,操作条件需高温高压,设备投资巨大,原料为不可再生资源,且环氧乙烷和另一路线的中间产物丙烯醛分别是易燃易爆或剧毒的危险品。由于发酵法生产1,3-丙二醇选择性高,操作条件温和,因此近年来受到特别的重视。
2,3-丁二醇(2,3-butanediol)作为发酵法生产1,3-丙二醇的一种副产物也是一种重要的化工原料。它是一种无色无味液体,可作为燃料,可用来制备聚合物、油墨、香水、防冻剂、熏蒸剂、增湿剂、软化剂、增塑剂、炸药以及药物的手性载体等。2,3-丁二醇还可作为一个很有价值的化工原料来合成其他化学品,如2,3-丁二醇脱水可产生甲乙酮,甲乙酮的应用相当广泛,再进一步脱水可形成1,3-丁二烯。2,3-丁二醇可通过Diels-Alder反应聚合生成苯乙烯。2,3-丁二醇与甲乙酮缩合并进行加氢反应生成辛烷,辛烷可用来产生高质量的飞行原料。2,3-丁二醇与乙酸反应生成2,3-丁二醇二乙酸酯,此酯类可添加到奶油中改善风味。但由于其在1,3-丙二醇发酵中产量较低,一般不作为产物进行分离、提纯。
目前,由甘油发酵生产1,3-丙二醇主要有以下几条途径:
1)采用肠道细菌厌氧条件下将甘油歧化为1,3-丙二醇(USP5254467,EP0373230 A1)。
2)克雷伯氏杆菌等厌氧菌在厌氧条件下发酵生产1,3-丙二醇(Ruch etal.Regulation of glycerol catabolism in Klebsiella aerogenes.J Bacteriol.1974,119(1):50-56;Streekstra et al.Overflow metabolism during anaeric growthof Klebsiella pneumoniae NCTC418 on glycerol and dihydroxyacetone inchemostat culture.Arch Microbiol.1987,147:268-275;Zeng et al.Pathwayanalysis of glycerol fermentation by Klebsiella pneumoniae:Regulation ofreducing equivalent balance and product formation.Enzyme Microbiol Technol.1993,15:770-779.)。
3)采用克雷伯氏杆菌在微氧条件下发酵生产1,3-丙二醇(王剑锋等,克雷伯氏菌微氧发酵生产1,3-丙二醇的研究,现代化工,2001,21(5):28-31。修志龙等,一种微生物微氧发酵生产1,3-丙二醇的方法,中国专利公开号:CN1348007)
4)采用克雷伯氏杆菌在厌氧条件下发酵产1,3-丙二醇和2,3-丁二醇(Biebl et al.Fermentation of glycerol to 1,3-propanediol and 2,3-butanediol.Appl Microbiol Biotechnol,1998,50:24-29)。
发明内容
本发明的目的是提供一种微生物两段发酵法由甘油生产1,3-丙二醇和2,3-丁二醇,其特征在于:所述由甘油生产1,3-丙二醇和2,3-丁二醇的方法是采用微生物前期厌氧、后期好氧的两段发酵新工艺。第一步厌氧发酵是将种子液加入含甘油浓度为20-80g/l的工业甘油或甘油发酵液的初始发酵培养基,在发酵温度30~40℃条件下利用厌氧菌进行厌氧发酵,在发酵的过程中通入0.2-0.4vvm的惰性气体氮气或二氧化碳,发酵开始3~5小时后流加900g/l甘油和90g/l葡萄糖混合溶液,其中甘油∶葡萄糖=10∶1(w/w);第二步进行好氧发酵,发酵温度仍保持在30~40℃,在发酵过程中通入空气或氧气,通气量为0.2-0.6vvm,发酵过程中同时流加500-1000g/l甘油、50-200g/l葡萄糖和3-5M氢氧化钠,其中甘油∶葡萄糖∶氢氧化钠=10∶1∶1(w/w/w)。发酵结束时通过常规的脱盐、蒸馏和真空精馏步骤将1,3-丙二醇和2,3-丁二醇分离、提纯,制备产品。
本发明的有益效果:采用前期厌氧后期好氧的两段发酵新工艺,可通过两段发酵过程中通气量的调节以及两段发酵转换时机的控制来调控1,3-丙二醇和2,3-丁二醇的产物浓度。提高产物浓度、得率和生产强度,并在得到产物1,3-丙二醇的同时将较高浓度的2,3-丁二醇分离,得到了产物2,3-丁二醇,从而有效提高原料利用率,降低生产成本。
具体实施方式
本发明提供一种微生物两段发酵法由甘油生产1,3-丙二醇和2,3-丁二醇,所述由甘油生产1,3-丙二醇和2,3-丁二醇的方法是采用微生物前期厌氧、后期好氧的两段发酵新工艺:第一步厌氧发酵是将种子液加入含甘油浓度为20-80g/l的工业甘油或甘油发酵液的初始发酵培养基中,在发酵温度30~40℃条件下利用厌氧菌-克雷伯氏杆菌进行厌氧发酵,在发酵的过程中通入0.2-0.4vvm的惰性气体氮气或二氧化碳等,发酵开始3~5小时后流加900g/l甘油和90g/l葡萄糖混合溶液,其中甘油∶葡萄糖=10∶1(w/w);第二步进行好氧发酵,发酵温度仍保持在30~40℃,在发酵过程中通入空气或氧气,通气量为0.2-0.6vvm,发酵过程中同时流加500-1000g/l甘油、50-200g/l葡萄糖和3-5M氢氧化钠,其中甘油∶葡萄糖∶氢氧化钠=10∶1∶1(w/w/w),发酵结束时通过常规的脱盐、蒸馏和真空精馏步骤将1,3-丙二醇和2,3-丁二醇分离、提纯,制备产品。
下面再举具体实施例对本发明予以进一步说明。
实例1:
(1)菌种:克雷伯氏杆菌(Klebsiella pneumoniae)M5al
(2)培养基
培养基组分 种子培养基 发酵培养基 微量元素溶液
(/l) (/l) (mg/l)
甘油 20g 20-80g ZnCl2 70
K2HPO4·3H2O 4.45g 2.225g MnCl2·4H2O 100
(NH4)2SO4 2.0g 2.0g H3BO3 60
KH2PO4 1.3g 0.65g CoCl2·6H2O 200
MgSO4·7H2O 0.2g 0.2g NiCl2·6H2O 25
酵母粉 1.0g 1.5g NiCl2·H2O 27.64
微量元素溶液 2ml 2ml Na2MoO4·2H2O 35
CaCO3 2.0g CuCl2·H2O 20
消泡剂 0.1ml CuSO4·5H2O 29.28
浓盐酸(37%) 0.9ml
(3)培养方式:
A.种子培养:
甘油管保藏的菌种转接至LB斜面,30℃下活化12小时。种子培养采用有氧培养,使用500ml三角瓶,装液量100ml。培养温度30℃,摇床转速140rpm。
B.发酵培养:发酵培养使用5L发酵罐,装液量4L,培养温度35℃,pH值7。发酵过程中用含浓度为50g/l的工业甘油的初始发酵培养基,发酵开始3.5小时后流加900g/l甘油和90g/l葡萄糖混合溶液,其中甘油∶葡萄糖=10∶1(w/w)。
(4)发酵方式
克雷伯氏杆菌发酵产1,3-丙二醇和2,3-丁二醇:发酵过程中发酵温度控制为35℃,前期厌氧发酵,通入氮气,通气量0.2vvm。在发酵进行至30小时进行后期好氧发酵,发酵过程中通入空气,通气量0.2vvm,发酵罐搅拌转速150rpm。。
(5)发酵结果如下:
发酵共进行68小时,残余甘油浓度8.3g/l,发酵液中含1,3-丙二醇61.7g/l,2-丁二醇20g/l,乳酸12g/l浓度,醋酸5g/l。1,3-丙二醇摩尔得率0.48,生产强度0.91g/(1h),2,3-丁二醇摩尔得率0.13。总二醇摩尔得率0.61
实例2:
(1)菌种:同实例1
(2)培养基:同实例1
(3)培养方式:
A.种子培养:
甘油管保藏的菌种转接至LB斜面,30℃下活化12小时。种子培养采用有氧培养,使用500ml三角瓶,装液量100ml。培养温度30℃,摇床转速140rpm。
B.发酵培养:发酵培养使用5L发酵罐,装液量4L,培养温度37℃,pH值7。发酵过程中用含浓度为30g/l工业甘油的初始发酵培养基,发酵开始4小时后流加800g/l甘油和80g/l葡萄糖混合溶液。其中甘油∶葡萄糖=10∶1(w/w)
(4)发酵方式:
克雷伯氏杆菌发酵产1,3-丙二醇和2,3-丁二醇:发酵过程中发酵温度控制为37℃,前期厌氧发酵,发酵过程中通入二氧化碳,通气量0.2vvm。在发酵进行至32小时进行后期好氧发酵,发酵过程中通入空气,通气量0.2vvm,发酵罐搅拌转速150rpm。。
(5)发酵结果如下:
发酵共进行68小时,残余甘油浓度10.7g/l,发酵液中含1,3-丙二醇69.6g/l,2,3-丁二醇16g/l,乳酸9.2g/l浓度,醋酸5.2g/l。1,3-丙二醇摩尔得率0.51,生产强度1.0g/(1h),2,3-丁二醇2,3-丁二醇0.09。总二醇摩尔得率0.6.
实例3:
(1)菌种:同实例1
(2)培养基:同实例1
(3)培养方式:
A.种子培养:
甘油管保藏的菌种转接至LB斜面,30℃下活化12小时。种子培养采用有氧培养,使用500ml三角瓶,装液量100ml。培养温度30℃,摇床转速140rpm。
B.发酵培养:发酵培养使用5L发酵罐,装液量4L,培养温度40℃,pH值7。发酵过程中用含80g/l甘油的甘油发酵液作为初始发酵培养基,发酵开始5小时后流加1000g/l甘油和100g/l葡萄糖混合溶液,其中甘油∶葡萄糖=10∶1(w/w)。
(4)发酵方式:
克雷伯氏杆菌发酵产1,3-丙二醇和2,3-丁二醇:发酵过程中发酵温度控制为40℃,前期厌氧发酵,发酵过程中通入氮气,通气量0.2vvm。在发酵进行至32小时进行后期好氧发酵,发酵过程中通入空气,通气量0.2vvm,发酵罐搅拌转速150rpm。
(5)发酵结果如下:
发酵共进行68小时,残余甘油浓度11.8g/l,发酵液中含1,3-丙二醇52.4g/l,2,3-丁二醇10.6g/l,乳酸7.4g/l浓度,醋酸3.7g/l。1,3-丙二醇摩尔得率0.53,生产强度0.77g/(1h),2,3-丁二醇摩尔得率0.09。总二醇摩尔得率0.62。
Claims (1)
1.一种微生物两段发酵法由甘油生产1,3-丙二醇和2,3-丁二醇,其特征在于:所述由甘油生产1,3-丙二醇和2,3-丁二醇的方法是采用微生物前期厌氧、后期好氧的两段发酵工艺:第一步厌氧发酵是将种子液加入含甘油浓度为20-80g/l的工业甘油或甘油发酵液的初始发酵培养基中,在发酵温度30~40℃条件下利用厌氧菌进行厌氧发酵,在发酵的过程中通入0.2-0.4vvm的惰性气体氮气或二氧化碳,发酵开始3~5小时后流加900g/l甘油和90g/l葡萄糖混合溶液,其中甘油∶葡萄糖=10∶1(w/w);第二步进行好氧发酵,发酵温度仍保持在30~40℃,在发酵过程中通入空气或氧气,通气量为0.2-0.6vvm,发酵过程中同时流加500-1000g/l甘油、50-200g/l葡萄糖和3-5M氢氧化钠,其中甘油∶葡萄糖∶氢氧化钠=10∶1∶1(w/w/w)。发酵结束时通过常规的脱盐、蒸馏和真空精馏步骤将1,3-丙二醇和2,3-丁二醇分离、提纯,制备产品。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 200410037692 CN1246465C (zh) | 2004-04-29 | 2004-04-29 | 微生物两段发酵法由甘油生产1,3-丙二醇和2,3-丁二醇 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 200410037692 CN1246465C (zh) | 2004-04-29 | 2004-04-29 | 微生物两段发酵法由甘油生产1,3-丙二醇和2,3-丁二醇 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1570123A true CN1570123A (zh) | 2005-01-26 |
CN1246465C CN1246465C (zh) | 2006-03-22 |
Family
ID=34481715
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN 200410037692 Expired - Lifetime CN1246465C (zh) | 2004-04-29 | 2004-04-29 | 微生物两段发酵法由甘油生产1,3-丙二醇和2,3-丁二醇 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN1246465C (zh) |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2006128381A1 (fr) * | 2005-06-03 | 2006-12-07 | Tsinghua University | Procede permettant de preparer le propanediol-1,3 en se servant de la glycerine en tant que sous-produit du carburant diesel biologique |
WO2006133637A1 (fr) * | 2005-06-17 | 2006-12-21 | Tsinghua University | Procédé de préparation du 1,3-propanediol et du 2,3-butanediol à partir de matériau d'amidon brut |
CN100463968C (zh) * | 2006-01-27 | 2009-02-25 | 华侨大学 | 一种由生物催化转化甘油制备1,3-丙二醇和二羟基丙酮的方法 |
CN101768610A (zh) * | 2010-03-05 | 2010-07-07 | 广东绿百多生物科技有限公司 | 一种高效培养乳酸菌和生产乳酸的方法 |
CN100999742B (zh) * | 2007-01-09 | 2011-05-11 | 清华大学 | 一种发酵生产1,3-丙二醇的高产工艺 |
US7985566B2 (en) | 2005-04-22 | 2011-07-26 | Mitsubishi Chemical Corporation | Biomass-resource-derived polyester and production process thereof |
CN1884560B (zh) * | 2006-05-31 | 2011-08-24 | 华东理工大学 | 一种发酵生产2,3-丁二醇的方法 |
US8324434B2 (en) | 2008-03-02 | 2012-12-04 | Dow Global Technologies, Llc | Hydrogenation process |
US8658408B2 (en) | 2008-06-09 | 2014-02-25 | Lanza Tech New Zealand Limited | Process for production of alcohols by microbial fermentation |
CN103890185A (zh) * | 2011-08-19 | 2014-06-25 | 基因组股份公司 | 用于生产2,4-戊二烯酸、丁二烯、丙烯、1,3-丁二醇和相关醇的微生物和方法 |
CN104611401A (zh) * | 2013-11-05 | 2015-05-13 | 中国石油化工股份有限公司 | 一种提高微生物发酵生产2,3-丁二醇的方法 |
-
2004
- 2004-04-29 CN CN 200410037692 patent/CN1246465C/zh not_active Expired - Lifetime
Cited By (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9080009B2 (en) | 2005-04-22 | 2015-07-14 | Mitsubishi Chemical Corporation | Biomass-resource-derived polyester and production process thereof |
US11732087B2 (en) | 2005-04-22 | 2023-08-22 | Mitsubishi Chemical Corporation | Biomass-resource-derived polyester and production process thereof |
US7985566B2 (en) | 2005-04-22 | 2011-07-26 | Mitsubishi Chemical Corporation | Biomass-resource-derived polyester and production process thereof |
US8021864B2 (en) | 2005-04-22 | 2011-09-20 | Mitsubishi Chemical Corporation | Biomass-resource-derived polyester and production process thereof |
US11560449B2 (en) | 2005-04-22 | 2023-01-24 | Mitsubishi Chemical Corporation | Biomass-resource-derived polyester and production process thereof |
US10870727B2 (en) | 2005-04-22 | 2020-12-22 | Mitsubishi Chemical Corporation | Biomass-resource-derived polyester and production process thereof |
US10287393B2 (en) | 2005-04-22 | 2019-05-14 | Mitsubishi Chemical Corporation | Biomass-resource-derived polyester and production process thereof |
WO2006128381A1 (fr) * | 2005-06-03 | 2006-12-07 | Tsinghua University | Procede permettant de preparer le propanediol-1,3 en se servant de la glycerine en tant que sous-produit du carburant diesel biologique |
WO2006133637A1 (fr) * | 2005-06-17 | 2006-12-21 | Tsinghua University | Procédé de préparation du 1,3-propanediol et du 2,3-butanediol à partir de matériau d'amidon brut |
US7968319B2 (en) | 2005-06-17 | 2011-06-28 | Tsinghua University | Method for producing 1,3-propanediol and 2,3-butanediol from raw starch material |
CN100463968C (zh) * | 2006-01-27 | 2009-02-25 | 华侨大学 | 一种由生物催化转化甘油制备1,3-丙二醇和二羟基丙酮的方法 |
CN1884560B (zh) * | 2006-05-31 | 2011-08-24 | 华东理工大学 | 一种发酵生产2,3-丁二醇的方法 |
CN100999742B (zh) * | 2007-01-09 | 2011-05-11 | 清华大学 | 一种发酵生产1,3-丙二醇的高产工艺 |
US8324434B2 (en) | 2008-03-02 | 2012-12-04 | Dow Global Technologies, Llc | Hydrogenation process |
US8658408B2 (en) | 2008-06-09 | 2014-02-25 | Lanza Tech New Zealand Limited | Process for production of alcohols by microbial fermentation |
CN101768610A (zh) * | 2010-03-05 | 2010-07-07 | 广东绿百多生物科技有限公司 | 一种高效培养乳酸菌和生产乳酸的方法 |
CN103890185A (zh) * | 2011-08-19 | 2014-06-25 | 基因组股份公司 | 用于生产2,4-戊二烯酸、丁二烯、丙烯、1,3-丁二醇和相关醇的微生物和方法 |
CN104611401A (zh) * | 2013-11-05 | 2015-05-13 | 中国石油化工股份有限公司 | 一种提高微生物发酵生产2,3-丁二醇的方法 |
CN104611401B (zh) * | 2013-11-05 | 2018-04-10 | 中国石油化工股份有限公司 | 一种提高微生物发酵生产2,3‑丁二醇的方法 |
Also Published As
Publication number | Publication date |
---|---|
CN1246465C (zh) | 2006-03-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP1897955B1 (en) | Method for preparing 1,3-propanediol and 2,3-butanediol from raw starch | |
CN1696297A (zh) | 利用生物柴油副产物甘油生产1,3-丙二醇的方法 | |
Deckwer | Microbial conversion of glycerol to 1, 3-propanediol | |
US8658408B2 (en) | Process for production of alcohols by microbial fermentation | |
CN100491537C (zh) | 微生物好氧发酵生产1,3-丙二醇的方法 | |
US20120045807A1 (en) | Process for producing chemicals using microbial fermentation of substrates comprising carbon monoxide | |
CN108603205B (zh) | 具有内部分隔器的低压分离器及其用途 | |
CN1246465C (zh) | 微生物两段发酵法由甘油生产1,3-丙二醇和2,3-丁二醇 | |
US10883121B2 (en) | Processes for fermentation and purification of value added products from gaseous substrates | |
CN101307336B (zh) | 构建基因工程菌发酵联产pdo、bdo和php的方法 | |
CN105705630B (zh) | 一种聚合级乳酸单体生产菌及其构建方法与乳酸制造技术 | |
JP6595508B2 (ja) | ピルビン酸塩由来生成物の生成及び制御のための発酵プロセス | |
CN102597252A (zh) | 生物燃料物质及生物化学物质的制备方法 | |
Vivek et al. | Production and applications of 1, 3-propanediol | |
CN106220512B (zh) | 一种制备丁二胺的方法 | |
de Cárdenas et al. | Production of organic acids via fermentation of sugars generated from lignocellulosic biomass | |
CN1304582C (zh) | 一种外源添加反丁烯二酸促进微生物合成1,3-丙二醇的方法 | |
Isar et al. | 1, 3-Propanediol: From Waste to Wardrobe | |
CN109337938B (zh) | 利用食烷菌发酵降解石蜡生产2-氧代己二酸的方法 | |
Fernández Naveira | Biofuels production (ethanol, butanol, hexanol) from renewable sources | |
WO2010079500A2 (en) | Aerobic production of 1,3-propanediol from crude glycerol from biodiesel process. | |
KR101453621B1 (ko) | 신규한 류코노스톡 메센터로이드 kpx-14 및 이를 이용한 d형 유산을 생산하는 방법 | |
AU2008321615B2 (en) | Novel bacteria and methods of use thereof | |
WO2009011645A1 (en) | Auto-immobilized co-fermentation |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CX01 | Expiry of patent term | ||
CX01 | Expiry of patent term |
Granted publication date: 20060322 |