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Abstract:
从海洋来源的假交替单胞菌(Pseudoalteromonas sp.)B1基因组中克隆1个褐藻胶裂解酶基因,并在大肠杆菌中实现异源表达,分离纯化重组酶并研究其酶学性质.通过Touch down PCR与热不对称交错PCR(thermal asymmetric interlaced PCR,TAIL-PCR)从假交替单胞菌B1基因组中扩增到1个褐藻胶裂解酶基因(B1SM),将目的基因插入到pGEX-4T-1载体,转化到宿主大肠杆菌(Escherichia coli,E.coli)BL21 (DE3).结果显示,从菌株B1克隆得到褐藻胶裂解酶基因B1SM,序列长度为1 005 bp,编码334个氨基酸,理论等电点为8.51,编码蛋白质的理论分子质量为37.13 kDa.重组褐藻胶裂解酶B1SM的最适pH和温度分别为8.0和25℃.该褐藻胶裂解酶在pH 7.0 ~9.0范围内,25℃下保温1h,仍剩余60%以上活力,pH稳定性较好.在最适pH 8.0和30℃下保温1h重组酶仍剩余60%以上活力.该研究为褐藻胶裂解酶基因B1SM在E.coli BL21(DE3)中实现了异源表达,为褐藻胶裂解酶的制备和研究奠定基础.
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食品与发酵工业
ISSN: 0253-990X
CN: 11-1802/TS
Year: 2019
Issue: 3
Volume: 45
Page: 77-82
Cited Count:
WoS CC Cited Count: 0
SCOPUS Cited Count: 1
ESI Highly Cited Papers on the List: 0 Unfold All
WanFang Cited Count: -1
Chinese Cited Count:
30 Days PV: 4
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