高产纤维素酶菌株的筛选、固态发酵条件优化及其酶学性质研究  

Screening and optimization of solid-state fermentation conditions of high cellulase-producing strain and its enzymatic property

作　　者：王彩衣 夏靖雯 庞冰瑜 覃丽媛 程忠 张婷 WANG Caiyi;XIA Jingwen;PANG Bingyu;QIN Liyuan;CHENG Zhong;ZHANG Ting(College of Food and Quality Engineering,Nanning University,Nanning 530200,China)

机构地区：[1]南宁学院食品与质量工程学院,广西南宁530200

出　　处：《中国酿造》2024年第9期105-111,共7页China Brewing

基　　金：南宁市“邕江计划”青年人才专项项目(RC20230202);2023年国家级大学生创新创业训练计划项目一般项目(202311549038);广西科技基地和人才专项(桂科AD24010031)。

摘　　要：该研究从实验室保藏的12株真菌菌株中筛选高产纤维素酶的菌株,并通过形态学观察及分子生物学技术对其进行菌种鉴定。以纤维素酶活力为评价指标,通过单因素和正交试验优化筛选菌株固态发酵产酶条件,并将其与已报道高产纤维素酶菌株的纤维素酶活力进行比较,最后对其粗纤维素酶的酶学性质进行初步研究。结果表明,筛选鉴定得到一株高产纤维素酶的棘孢木霉(Trichoderma asperellum)BW-6,其滤纸酶(FPase)和羧甲基纤维素酶(CMCase)活力分别达到1.99 U/g和7.02 U/g,其最佳固态发酵产酶条件为:以麦麸+甘蔗渣+稻秆(2.0 g+1.5 g+1.5 g)为固态发酵基质,基础盐溶液培养基中以0.5%蛋白胨为氮源,发酵液添加量6 mL,26℃静置培养5 d。在此条件下,滤纸酶活力和羧甲基纤维素酶活力分别达到2.81 U/g和10.15 U/g,分别是优化前的1.41倍和1.46倍,且极显著高于同等条件下草酸青霉(Penicillium oxalicum)EU2101和EU2106的纤维素酶活力(P<0.01),表明菌株BW-6具有工业化应用潜能。此外,菌株BW-6所产纤维素酶的最适反应温度和pH分别为50℃和4.0,在45℃和pH 5.5~6.0条件下稳定。In this study,the high cellulase-producing strain was screened from 12 fungi preserved in the laboratory,and identified by morphological observation and molecular biology techniques.Using cellulase activity as the evaluation index,the solid-state fermentation conditions of the screened strain were optimized by single factor tests and orthogonal tests,and the cellulase activity was compared with that of the reported high cellulase-producing strains.Finally,the enzymatic properties of the crude cellulase were preliminarily investigated.The results showed that a high cellulase-producing strain of Trichoderma asperellum BW-6 was screened and identified,with the filter paperase(FPase)and carboxymethyl cellulase(CMCase)activities reached 1.99 U/g and 7.02 U/g,respectively,and the optimal solid-state fermentation conditions were as follows:wheat bran+bagasse+rice straw(2.0 g+1.5 g+1.5 g)as solid-state fermentation substrate,0.5%peptone in basic salt solution medium as nitrogen source,fermentation broth addition 6 ml,and static culture at 26℃for 5 d.Under these conditions,the filter paperase and the carboxymethyl cellulase activities reached 2.81 U/g and 10.15 U/g,respectively,which were 1.41 and 1.46 times that of before optimization,and were extremely higher than that of Penicillium oxalicum EU2101 and EU2106 under the same conditions(p<0.01),indicating that the strain BW-6 had the potential for industrial application.In addition,the optimal reaction temperature and pH of the cellulase produced by strain BW-6 were 50℃and 4.0,respectively,and it was stable at 45℃and pH 5.5-6.0.

关 键 词：棘孢木霉 固态发酵 纤维素酶 发酵条件 正交试验 酶学性质 

分 类 号：Q815[生物学—生物工程]