粘性丝孢酵母的诱变及以玉米秸秆水解液为原料产油条件优化
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摘要
对粘性丝胞酵母进行紫外诱变,获得一株产油率较高的菌株,较原菌株提高了1.53倍。将该菌株接种于用1%硫酸和酶水解处理并浓缩至还原糖浓度为5%的玉米秸秆水解液中培养,生长较好。通过四因素三水平正交实验,确定培养条件为初始pH值7.0、接种量1%、发酵温度30℃、发酵时间5 d时产油率最高。对最佳产油条件进行验证,测得油脂含量为21.3%。从而为利用农业废弃物大规模生产微生物油脂提供了试验数据。气相色谱-质谱联用分析仪显示油脂脂肪酸组成为棕榈酸28.36%,油酸55.86%,10-十八烯酸9.23%,硬脂酸6.70%,可以作为原料生产生物柴油。
A strain with high oil yield rate was obtained by UV mutagenesis on Trichosporon mucoides, which was 1.53 times higher than that of the original strain. The strain was cultured in the corn straw hydrolysate which was treated with 1% sulfuric acid and enzymatic hydrolysis and condensed to 5% reducing sugar. It was found that the strain grew well. Through the orthogonal experiment of four factors and three levels, the oil yield rate was the highest when the culture conditions including the initial p H of 7.0, the inoculum concentration of 1%, the fermentation temperature of 30℃ and the fermentation time of 5 days. Through verifying the optimal condition of oil yield, the lipid content was measured as 21.3%. Thus, the experiment data might be provided for large-scale production of microbial oil by using agricultural wastes. The analysis of gas-chromatograph and mass spectrometry showed that the oil fatty acid compositions were 28.36% of hexadecanoic acid, 55.86% of oleic acid, 9.23% of10-octadecenoic acid, 6.70% of stearic acid, which could be used as raw material to produce biodiesel.
A strain with high oil yield rate was obtained by UV mutagenesis on Trichosporon mucoides, which was 1.53 times higher than that of the original strain. The strain was cultured in the corn straw hydrolysate which was treated with 1% sulfuric acid and enzymatic hydrolysis and condensed to 5% reducing sugar. It was found that the strain grew well. Through the orthogonal experiment of four factors and three levels, the oil yield rate was the highest when the culture conditions including the initial p H of 7.0, the inoculum concentration of 1%, the fermentation temperature of 30℃ and the fermentation time of 5 days. Through verifying the optimal condition of oil yield, the lipid content was measured as 21.3%. Thus, the experiment data might be provided for large-scale production of microbial oil by using agricultural wastes. The analysis of gas-chromatograph and mass spectrometry showed that the oil fatty acid compositions were 28.36% of hexadecanoic acid, 55.86% of oleic acid, 9.23% of10-octadecenoic acid, 6.70% of stearic acid, which could be used as raw material to produce biodiesel.
引文
Azócar L.,Heipieper H.J.,and Navia R.,2010,Biotechnological processes for biodiesel production using alternative oils,Appl.Microbiol.Biotechnol.,88(3):621-636
Alberto A.,Stefano R.,Alan L.,and Maddalena R.,2012,Candida Freyschussii:an oleaginous yeast producing lipids from glycerol,Chemical Engineering,27(12):139-144
Li X.,Li D.M.,Guo X.M.,Ge X.,and Shen P.H.,2010,Research progress on industrialized application of microbial oils,Zhongguo Youzhi(China Oils and Fats),35(1l):10-13(李翔,黎冬明,郭晓敏,葛霞,申鹏华,2010,微生物油脂产业化应用研究进展,中国油脂,35(1l):10-13)
Liu B.,and Zhao Z.B.,2007,Biodiesel production by direct methanolysis of oleaginous microbial biomass,J.Chem.Technol.Biotechnol.,82(8):775-780
Liu H.J.,Zhang J.A.,Mo Y.X.,Li J.,Zhou U.J.,and Cheng K.K.,2008,Biodiesel production with microbial lipid,Huaxue Fanying Gongcheng Yu Gongyi(Chemical Reaction Engineering and Technology),24(3):263-267(刘宏娟,张建安,墨玉欣,李建,周玉杰,程可可,2008,利用微生物油脂制备生物柴油,化学反应工程与工艺,24(3):263-267)
Lin Y.,and Tanaka S.,2006,Ethanol fermentation form biomass resources:current state and prospects,Appl.Microbiol.Biotechnol.,69(6):627-642
Li X.S.,Lu B.S.,and Jin S.X.,2011,Application of enzyme in production of microbial oils fermented by two-strains with corn stalk,Zhongnan Daxue Xuebao(Journal of Central South University(Science and Technology)),42(6):1560-1566(李新社,陆步诗,金双喜,2011,酶制剂在玉米秸秆双菌种固态发酵生产微生物油脂中的应用,中南大学学报(自然科学版),42(6):1560-1566)
Meng X.,Yang J.,Xu X.,Zhang L.,Nie Q.J.,and Xian M.,2009,Biodiesel production from oleaginous microorganisms,Renewable Energy,34(1):1-5
Pu F.L.,Sun W.F.,and Che Z.M.,2016,Research and development progress of functional oils,Liangshi Yu Youzhi(Cereals and Oils),29(8):5-8(蒲凤琳,孙伟峰,车振明,2016,功能性油脂研究与开发进展,粮食与油脂,29(8):5-8)
Wang X.,Hu J.D.,Zhang X.J.,and Yang H.T.,2011,Studies on lipids production by Trichoderma spp.from lignocellulosic materials,Zhongguo Niangzao(China Brewing),30(12):1-4(王雪,扈进东,张新建,杨合同,2011,木霉菌株利用作物秸秆产油脂研究进展,中国酿造,30(12):1-4)
Zhu L.Y.,Zong M.H.,and Wu H.,2008,Efficient lipid production with Trichosporon fermentans and its use for biodiesel preparation,Bioresour.Technol.,99(16):7881-7885
Zhang G.L.,Du W.,and Liu D.H.,2013,Research progress in biodiesel production from yeast lipid,Huagong Jinzhan(Chemical Industry and Engineering Progress),32(4):791-797(张国玲,杜伟,刘德华,2013,酵母油脂及用于生物柴油制备研究进展,化工进展,32(4):791-797)
Alberto A.,Stefano R.,Alan L.,and Maddalena R.,2012,Candida Freyschussii:an oleaginous yeast producing lipids from glycerol,Chemical Engineering,27(12):139-144
Li X.,Li D.M.,Guo X.M.,Ge X.,and Shen P.H.,2010,Research progress on industrialized application of microbial oils,Zhongguo Youzhi(China Oils and Fats),35(1l):10-13(李翔,黎冬明,郭晓敏,葛霞,申鹏华,2010,微生物油脂产业化应用研究进展,中国油脂,35(1l):10-13)
Liu B.,and Zhao Z.B.,2007,Biodiesel production by direct methanolysis of oleaginous microbial biomass,J.Chem.Technol.Biotechnol.,82(8):775-780
Liu H.J.,Zhang J.A.,Mo Y.X.,Li J.,Zhou U.J.,and Cheng K.K.,2008,Biodiesel production with microbial lipid,Huaxue Fanying Gongcheng Yu Gongyi(Chemical Reaction Engineering and Technology),24(3):263-267(刘宏娟,张建安,墨玉欣,李建,周玉杰,程可可,2008,利用微生物油脂制备生物柴油,化学反应工程与工艺,24(3):263-267)
Lin Y.,and Tanaka S.,2006,Ethanol fermentation form biomass resources:current state and prospects,Appl.Microbiol.Biotechnol.,69(6):627-642
Li X.S.,Lu B.S.,and Jin S.X.,2011,Application of enzyme in production of microbial oils fermented by two-strains with corn stalk,Zhongnan Daxue Xuebao(Journal of Central South University(Science and Technology)),42(6):1560-1566(李新社,陆步诗,金双喜,2011,酶制剂在玉米秸秆双菌种固态发酵生产微生物油脂中的应用,中南大学学报(自然科学版),42(6):1560-1566)
Meng X.,Yang J.,Xu X.,Zhang L.,Nie Q.J.,and Xian M.,2009,Biodiesel production from oleaginous microorganisms,Renewable Energy,34(1):1-5
Pu F.L.,Sun W.F.,and Che Z.M.,2016,Research and development progress of functional oils,Liangshi Yu Youzhi(Cereals and Oils),29(8):5-8(蒲凤琳,孙伟峰,车振明,2016,功能性油脂研究与开发进展,粮食与油脂,29(8):5-8)
Wang X.,Hu J.D.,Zhang X.J.,and Yang H.T.,2011,Studies on lipids production by Trichoderma spp.from lignocellulosic materials,Zhongguo Niangzao(China Brewing),30(12):1-4(王雪,扈进东,张新建,杨合同,2011,木霉菌株利用作物秸秆产油脂研究进展,中国酿造,30(12):1-4)
Zhu L.Y.,Zong M.H.,and Wu H.,2008,Efficient lipid production with Trichosporon fermentans and its use for biodiesel preparation,Bioresour.Technol.,99(16):7881-7885
Zhang G.L.,Du W.,and Liu D.H.,2013,Research progress in biodiesel production from yeast lipid,Huagong Jinzhan(Chemical Industry and Engineering Progress),32(4):791-797(张国玲,杜伟,刘德华,2013,酵母油脂及用于生物柴油制备研究进展,化工进展,32(4):791-797)