匍枝根霉变种reflexus TP - 02纤维素酶基因EG1在大肠杆菌中的克隆与表达
Cloning and expression of cellulase gene EG1 from Rhizopus stolonifer var. reflexus TP-02 in Escherichia coli.
作者信息
Tang Bin, Pan Haibo, Zhang Qingqing, Ding Lixia
机构信息
Department of Biochemical Engineering, Anhui University of Technology and Science, Wuhu 241000, China.
出版信息
Bioresour Technol. 2009 Dec;100(23):6129-32. doi: 10.1016/j.biortech.2009.06.091. Epub 2009 Jul 28.
A novel gene, EG encoding enzymes involved in carboxymethyl cellulose (CMC) degradation was isolated, sequenced from the filamentous fungus Rhizopus stolonifer var. reflexus TP-02, and expressed in Escherichia coli BL21. The results showed that the gene amplified from the cDNA of the strain could be classified as the family of endoglucanase. During the fermentation process, the maximum endoglucanase activity (i.e. 0.715 IU/ml) of the recombinant bacteria was obtained at 36 h. The SDS-PAGE analysis on purified samples showed that a band with apparent molecular weight of about 40 kDa was detected after staining with Coomassie brilliant blue.
从丝状真菌匍枝根霉变种TP-02中分离出一个编码参与羧甲基纤维素（CMC）降解酶的新基因EG，对其进行测序，并在大肠杆菌BL21中表达。结果表明，从该菌株cDNA中扩增出的基因可归类为内切葡聚糖酶家族。在发酵过程中，重组菌在36 h时获得最大内切葡聚糖酶活性（即0.715 IU/ml）。对纯化样品进行的SDS-PAGE分析表明，用考马斯亮蓝染色后检测到一条表观分子量约为40 kDa的条带。
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