高效降解纤维素微生物产酶条件优化及复合菌群的构建
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摘要
以羧甲基纤维素钠(CMC-Na)为唯一碳源,从白蚁肠道中分离出具有高效降解纤维素能力的菌株,通过16s rDNA鉴定种属关系,三株菌株命名为2号、4号、5号,2、5号为蜡样芽孢杆菌、4号为链球菌。通过条件优化,确定最佳发酵条件为:CMC-Na为碳源、牛肉膏为氮源、pH值为7、37℃、发酵时间24 h、2%接种量。随机组合不同菌株构建复合菌群,复合菌群产酶能力最强的为4号+5号菌株,酶活达3.67 U/mL。
Using sodium carboxymethyl cellulose sodium(CMC-Na) as the sole carbon source, the strains with the ability to degrade cellulose were isolated from the intestinal tract of termites. The species relationship was identified by 16 s rDNA. The three strains were named No. 2, No. 4 and No. 5. No. 2 and No.5 are Bacillus cereus, No.4 is Streptococcus. Through the optimization of the conditions,the optimum fermentation conditions were determined as follows: CMC-Na as carbon source, beef paste as nitrogen source, pH 7, 37 ℃,fermentation time 24 h, inoculum size 2%. The complex microbiotawas constructed by random combination of different strains, and the enzyme production ability of the compound microbiota(No.4+No.5) was the strongest, and the enzyme activity was 3.67 U/mL.
Using sodium carboxymethyl cellulose sodium(CMC-Na) as the sole carbon source, the strains with the ability to degrade cellulose were isolated from the intestinal tract of termites. The species relationship was identified by 16 s rDNA. The three strains were named No. 2, No. 4 and No. 5. No. 2 and No.5 are Bacillus cereus, No.4 is Streptococcus. Through the optimization of the conditions,the optimum fermentation conditions were determined as follows: CMC-Na as carbon source, beef paste as nitrogen source, pH 7, 37 ℃,fermentation time 24 h, inoculum size 2%. The complex microbiotawas constructed by random combination of different strains, and the enzyme production ability of the compound microbiota(No.4+No.5) was the strongest, and the enzyme activity was 3.67 U/mL.
引文
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[7]冯冲,苏丽娟,张洋,等.白蚁肠道内纤维素酶产生菌的分离与鉴定[J].河南农业大学学报,2013,47(5):587-591.
[8]李俭,文戈,薛高尚,等.白蚁肠道纤维素酶菌株筛选及生长条件研究[J].江西农业学报,2012,24(8):81-84.
[9]何刚强,堵国成,刘立明,等.从白蚁中分离筛选纤维素分解菌及其产酶性质[J].食品与生物技术学报,2009,28(3):352-355.
[10]高云航,王巍,李秋菊,等.白蚁肠道木质素及纤维素分解菌的分离鉴定及产酶条件优化[J].中山大学学报(自然科学版),2013,52(2):84-89.
[2]Collins N M,Wood T G.Termites and atmospheric gas production[J].Science,1984,224(4644):84-86.
[3]Lo N,Tokuda G,Watanabe H,et al.Evidence from multiple gene sequences indicates that termites evolved from wood-feeding cockroaches[J].Current Biology,2000,10(13):801-804.
[4]Ni J F,Takehara M,Watanabe H.Heterologous overexpression of a mutant termite cellulase gene inescherichia coliby DNA shuffling of four orthologous parental cDNAs[J].Bioscience,Biotechnology,and Biochemistry,2005,69(9):1711-1720.
[5]Li XH,Yang HJ,Roy B,et al.The most stirring technology in future:cellulase enzyme and biomass utilization[J].African Journal of Biotechnology,2009,8(11):2418-2422.
[6]黄耀华,曹志方,王学梅,等.海南树栖白蚁肠道中高产纤维素酶菌株的筛选[J].热带生物学报,2013,4(4):342-346.
[7]冯冲,苏丽娟,张洋,等.白蚁肠道内纤维素酶产生菌的分离与鉴定[J].河南农业大学学报,2013,47(5):587-591.
[8]李俭,文戈,薛高尚,等.白蚁肠道纤维素酶菌株筛选及生长条件研究[J].江西农业学报,2012,24(8):81-84.
[9]何刚强,堵国成,刘立明,等.从白蚁中分离筛选纤维素分解菌及其产酶性质[J].食品与生物技术学报,2009,28(3):352-355.
[10]高云航,王巍,李秋菊,等.白蚁肠道木质素及纤维素分解菌的分离鉴定及产酶条件优化[J].中山大学学报(自然科学版),2013,52(2):84-89.