拜氏梭菌Clostridium beijerinckii NCIMB 8052发酵玉米皮生产丁醇

玉米皮作为玉米淀粉加工的副产物,是一种可用于生产液体燃料的潜在廉价优质的生物质资源。本文以玉米皮为原料,对拜氏梭菌发酵生产丁醇进行了研究。实验结果表明,玉米皮首先在最优的预处理温度140℃下使用0.5%硫酸水溶液以固液比1∶8处理20 min,再添加200 IU/g底物糖化酶、1.0 IU/g底物木聚糖酶进行酶解,可以使原料中的淀粉和半纤维素转化为可发酵糖,此时水解液中的总糖浓度为50.46 g/L。然后使用1.0%的活性炭对水解液进行脱毒处理以去除发酵抑制物,再进行丁醇发酵,丁醇产量为9.72 g/L,总溶剂产量可达14.09 g/L,糖醇转化率为35.1%。上述研究结果证明玉米皮作为一种粮食加工废弃物用于液体燃料丁醇的生产在技术上是完全可行的。     

丙酮丁醇梭菌发酵菊芋汁生产丁醇

对丙酮丁醇梭菌Clostridium acetobutylicum L7发酵菊芋汁酸水解液生产丁醇进行了初步研究。实验结果表明,以该水解液为底物生产丁醇,不需要添加氮源和生长因子。当水解液初始糖浓度为48.36 g/L时,其发酵性能与以果糖为碳源的对照组基本相同,发酵终点丁醇浓度为8.67 g/L,丁醇、丙酮和乙醇的比例为0.58∶0.36∶0.06,但与以葡萄糖为碳源的对照组相比,发酵时间明显延长,表明该菌株葡萄糖转运能力强于果糖。当水解液初始糖浓度提高到62.87 g/L时,发酵终点残糖浓度从3.09 g/L增加到3.26 g/L,但丁醇浓度却提高到11.21 g/L,丁醇、丙酮和乙醇的比例相应为0.64∶0.29∶0.05,表明适量糖过剩有助于C.acetobutylicum L7胞内代谢从丙酮合成向丁醇合成途径调节;继续提高水解液初始糖浓度,发酵终点残糖浓度迅速升高,丁醇生产的技术经济指标受到明显影响。     

丙酮丁醇梭菌XY16丁醇发酵特性

以诱变选育的1株突变菌株丙酮丁醇梭菌XY16为对象,对影响该菌发酵特性的相关因素(N源、生长因子、热激)进行研究。结果显示:无机N源乙酸铵比其他N源更有利于丙酮丁醇的发酵,玉米浆或玉米蛋白可以直接替代生长因子进行丙酮丁醇发酵,热激可以提高总溶剂产量,最高可以达到21.28 g/L。该菌还可以同时利用葡萄糖和木糖,当葡萄糖利用完后,木糖才能被有效利用。     

稻草酶法水解液的丙酮丁醇发酵

利用丙酮丁醇梭菌Ｃ３７５菌株发酵稻草酶法水解液,分别研究了氮源、生长因子ＰＨ等因素对发酵的影响。结果表明,在稻草水解液还原糖浓度为４．２８％时,总溶剂为１２．８ｇ／Ｌ,溶剂组成：丁醇：丙酮：乙醇＝６５．８：２３．８：１０．４,溶剂生成率为２９．９％。     

丙酮丁醇发酵菌的分子遗传改造

丙酮丁醇梭菌及拜氏梭菌是重要的ABE（丙酮、丁醇和乙醇）工业生产菌株,其发酵产物中的丙酮和丁醇均为重要的化工原料,汽车发动机试验证明丁醇还是一种性能优于乙醇的极具潜力的生物燃料和燃料添加剂。随着新生物技术的不断发展及工业生产的需求,遗传工程改造不断应用于丙酮丁醇生产菌株。在前人研究及工业实践的基础上,对丙酮丁醇生产菌株的遗传特性及其分子遗传改造取得的进展进行了详细概述。     

高丁醇比丙酮丁醇梭菌的选育与应用

设计了专一性分离方法,从土样中分离了多株能产生溶剂的梭苗,经多次单细胞分离、纯化,再经亚硝基胍和甲基磺酸乙酯诱变和抗性筛选,获得几株高丁醇的丙酮丁醇梭菌。对高产菌株的性状稳定性、发酵过程、混合原料应用、温度的影响进行了研究。结果证明菌株性状稳定,丁醇产量为总溶剂的７０％;过程为典型的丙酮丁醇发酵,对温度可耐受到３９－４０℃;能利用玉米和薯干,玉米和高梁进行正常发酵。菌株已在百吨生产罐,连续应用一年     

木薯发酵产丁醇的研究

对丙酮丁醇梭菌发酵木薯产溶剂进行研究,分别考察了N源、木薯含量、酶处理条件和培养基pH对发酵产丁醇的影响。结果表明：最佳的产丁醇发酵培养基为木薯粉120g／L,乙酸铵6g／L;木薯粉先用高温淀粉酶按酶量20U／g、90℃水解60min,再糊化30min;发酵初始pH为6．0,发酵96h。在此条件下,5L发酵罐中丁醇产量达到13．5g／L,总溶剂达到22．8g／L。     

丁酸胁迫耦联丙酮丁醇梭菌-酿酒酵母混合培养强化丁醇发酵性能

为改善丁醇发酵性能,提出丁酸胁迫与丙酮丁醇梭菌-酿酒酵母混合培养体系协同作用的新型丁醇发酵优化控制策略.7L发酵罐中,在溶剂生产期(24 h)添加4.0 g/L-broth的丁酸浓缩液和0.2 g-DCW/L-broth的酿酒酵母进行发酵,丁醇浓度、丁醇/丙酮比和总溶剂生产效率与对照相比分别提高35％、43％和79％,达到15.74 g/L、2.83和0.52 g/L/h的最高水平.若将精馏后溶剂混合物作为高效柴油添加剂,柴油添加剂中B∶A∶E比例可达74∶17∶9(w/w)的高水平,产品质量获得显著改善.试验及分析阐明该优化控制策略可大幅诱发赖氨酸的分泌及在梭菌中的吸收/利用,提高梭菌对高丁醇浓度环境的耐受能力,促进丁醇合成;可强化梭菌对底物利用的竞争能力、提高电子往复穿梭传递系统中还原力再生速率、产生更多用于丁醇合成的NADH.两者的协同作用大幅提高了丁醇发酵的整体性能.     

萃取耦合发酵工艺生产丁醇研究

萃取耦合发酵可有效减弱产物抑制和提高底物利用效率,本文就萃取耦合发酵生产丁醇工艺中的萃取剂的选择、萃取剂加入量、底物浓度等发酵条件进行了研究。结果表明:最佳萃取剂为大豆油生物柴油,油水比为3∶5,发酵过程无需搅拌,静置发酵为宜,在发酵之初加入萃取剂。分别以玉米和木薯为发酵底物,确定其最适底物浓度为100 g/L,以玉米为原料萃取耦合发酵中丁醇和总溶剂产量分别为18.17 g/L和29.31 g/L。以木薯为原料萃取耦合发酵生产丁醇及总溶剂产量比传统发酵分别提高了48.69%和51.80%。     

蔗渣水解液发酵乙醇的研究

研究了酵母（Ｐｉｃｈｉａｓｔｉｐｉｔｉｓ）Ｙ７１２４在限制供氧条件下尽管反应初期葡萄糖消耗速率大于木糖,但在一定时间后,葡萄糖的消耗速率变慢,而木糖消耗速率变快直至耗尽的现象。建立了气升柱以Ｐ．Ｓｔｉｐｉｔｉｓ转化木糖为目的和以溢流柱Ｓａｃｈａｒｏｍｙｃｅｓｃｅｒｉｖｉｓｉａｅ转化残留葡萄糖为目的的串联发酵乙醇工艺,即流加５倍浓缩的蔗渣水解液,Ｄ＝０.１ｈ^－１,还原糖总利用率为９７.２％,酒精浓度为４６.５ｇ／Ｌ,生产率为４.１ｇ／Ｌ·ｈ。     

Butanol production by Clostridium beijerinckii. Part I: use of acid and enzyme hydrolyzed corn fiber

Fermentation of sulfuric acid treated corn fiber hydrolysate (SACFH) inhibited cell growth and butanol production (1.7 ± 0.2 g/L acetone butanol ethanol or ABE) by Clostridium beijerinckii BA101. Treatment of SACFH with XAD-4 resin removed some of the inhibitors resulting in the production of 9.3 ± 0.5 g/L ABE and a yield of 0.39 ± 0.015. Fermentation of enzyme treated corn fiber hydrolysate (ETCFH) did not reveal any cell inhibition and resulted in the production of 8.6 ± 1.0 g/L ABE and used 24.6 g/L total sugars. ABE production from fermentation of 25 g/L glucose and 25 g/L xylose was 9.9 ± 0.4 and 9.6 ± 0.4 g/L, respectively, suggesting that the culture was able to utilize xylose as efficiently as glucose. Production of only 9.3 ± 0.5 g/L ABE (compared with 17.7 g/L ABE from fermentation of 55 g/L glucose-control) from the XAD-4 treated SACFH suggested that some fermentation inhibitors may still be present following treatment. It is suggested that inhibitory components be completely removed from the SACFH prior to fermentation with C. beijerinckii BA101. In our fermentations, an ABE yield ranging from 0.35 to 0.39 was obtained, which is higher than reported by the other investigators.     

Butanol production from agricultural residues: Impact of degradation products on Clostridium beijerinckii growth and butanol fermentation

During pretreatment and hydrolysis of fiber-rich agricultural biomass, compounds such as salts, furfural, hydroxymethyl furfural (HMF), acetic, ferulic, glucuronic, rho-coumaric acids, and phenolic compounds are produced. Clostridium beijerinckii BA101 can utilize the individual sugars present in lignocellulosic [e.g., corn fiber, distillers dry grain solubles (DDGS), etc] hydrolysates such as cellobiose, glucose, mannose, arabinose, and xylose. In these studies we investigated the effect of some of the lignocellulosic hydrolysate inhibitors associated with C. beijerinckii BA101 growth and acetone-butanol-ethanol (ABE) production. When 0.3 g/L rho-coumaric and ferulic acids were introduced into the fermentation medium, growth and ABE production by C. beijerinckii BA101 decreased significantly. Furfural and HMF are not inhibitory to C. beijerinckii BA101; rather they have stimulatory effect on the growth of the microorganism and ABE production.     

一株拜氏梭菌利用甘蔗废糖蜜发酵生产丙酮丁醇

以甘蔗废糖蜜作为原料,利用Clostridium beijerinckii DSM 6422菌株进行丙酮丁醇发酵的初步研究.结果表明:采用H2SO4预处理糖蜜,初糖质量浓度60 g/L,(NH4)2SO4 2g/L,CaCO3 10 g/L,温度30℃,pH 5.5～7.0,接种量6％(体积分数),在5L发酵罐中发酵培养96 h,总溶剂产量为16.17 g/L,其中丁醇质量浓度为10.07 g/L,总溶剂产率为30.2％,糖利用率为89.3％.     

Production of biobutanol from acid-pretreated corncob using Clostridium beijerinckii TISTR 1461: Process optimization studies

Corncob is a potential feedstock in Thailand that can be used for fermentable sugar production through dilute sulfuric acid pretreatment and enzymatic hydrolysis. To recover high amounts of monomeric sugars from corncob, the sulfuric pretreatment conditions were optimized by using response surface methodology with three independent variables: sulfuric acid concentration, temperature, and time. The highest response of total sugars, 48.84 g/L, was found at 122.78°C, 4.65 min, and 2.82% (v/v) H₂SO₄. With these conditions, total sugars from the confirmation experiment were 46.29 g/L, with 5.51% error from the predicted value. The hydrolysate was used as a substrate for acetone–butanol–ethanol fermentation to evaluate its potential for microbial growth. The simultaneous saccharification and fermentation (SSF) showed that C. beijerinckii TISTR 1461 can generate acetone–butanol–ethanol products at 11.64 g/L (5.29 g/L acetone, 6.26 g/L butanol, and 0.09 g/L ethanol) instantly using sugars from the hydrolysed corncob with Novozymes 50013 cellulase enzyme without an overliming process.     

Development of a cost-effective glucose-corn steep medium for production of butanol by Clostridium beijerinckii

Corn steep water (CSW) medium (1.6% solids plus 6% glucose) was evaluated for growth and butanol production by Clostridium beijerinckii NCIMB 8052 wild-type and hyper-amylolytic, hyper-butanol-producing mutant strain BA101. CSW alone was not a suitable substrate, whereas addition of glucose supported growth and butanol production by both strains. In a batch-scale fermentation using an optimized 6% glucose-1.6% solids CSW medium, C. beijerinckii NCIMB 8052 and strain BA101 produced 10.7 g L⁻¹ and 14.5 g L⁻¹ of butanol, respectively. The total solvents (acetone, butanol, and ethanol) produced by C. beijerinckii NCIMB 8052 and strain BA101 were 14 g L⁻¹ and 20 g L⁻¹, respectively. Initial fermentation in small-scale flasks containing 6% maltodextrin-1.6% solids concentration CSW medium resulted in 6 g L⁻¹ and 12.6 g L⁻¹ of butanol production by C. beijerinckii NCIMB 8052 and strain BA101, respectively. CSW can serve as an economic source of nitrogen, vitamins, amino acids, minerals, and other nutrients. Thus, it is feasible to use 6% glucose-1.6% solids CSW medium in place of semi-defined P2 medium. Received 9 February 1998/ Accepted in revised form 1 September 1998