一株耐热菌的产酶研究及其羧酸酯酶基因的克隆-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Enzyme Production of a Thermophilic Strain and Cloning of Its Carboxylesterase Gene

作者:: 朱艳冰1; 王国红2; 陈少娟1; 刘光明1; (1.集美大学生物工程学院,福建厦门 361021;2.新乡医学院基础医学院,河南新乡 453003)

Author(s):: ZHU Yan-bing1; WANG Guo-hong2; CHEN Shao-juan1; LIU Guang-ming1; (1.School of Biotechnology Engineering,Jimei University,Xiamen 361021,China2.School of Preclinical Medicine,Xinxiang Medical University,Xinxiang 453003,China)

Keywords:: thermophile; qualitative enzyme production; carboxylesterase gene

摘要:: 将一株近海温泉耐热菌ZH1的16S rDNA克隆至pUCm–T载体进行测序，测序结果在NCBI上进行BLAST分析，并构建了系统发育树，将该菌株归属为Geobacillus spZH1.在65 ℃条件下的产酶定性实验结果表明，该菌株产脂肪酶、木聚糖酶和碱性磷酸酶.以菌株ZH1的基因组DNA为模板，使用羧酸酯酶简并引物进行PCR扩增，将目的基因克隆至pUCm–T载体后进行测序，得到了741 bp的DNA片段.BLAST分析显示，目的基因预测编码羧酸酯酶，包含246个氨基酸

Abstract:: The 16S rDNA from an inshore hot spring thermophilic strain ZH1 was cloned into pUCm-T vector and then sequenced.By analyzing the sequence against GenBank database with BLAST and constructing the phylogenetic tree,this strain was assigned to Geobacillus sp.ZH1.Qualitative analysis of its enzyme production at 65 ℃ showed that this strain could produce lipase,xylanase and alkaline phosphatase.To clone the carboxylesterase gene,it was amplified with the bacterial genomic DNA using a pair of degenerate primers.The target gene was cloned into pUCm-T vector and then sequenced.The obtained 741 bp DNA fragment was predicted to encode a carboxylesterase containing 246 amino acid residues with BLAST