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脱硫菌Rhodococcus sp.LY822专一性脱硫活性及相关基因的研究
Authors
刘会洲
姜声华
+3 more
李信
熊小超
白雪晶
Publication date
1 January 2008
Publisher
Abstract
以专一性脱硫菌Rhodococcus sp.LY822质粒DNA为模板,利用已知的脱硫基因序列设计引物,PCR扩增得到了3个脱硫基因片段dszA,dszB和dszC.构建了相应的表达质粒pETA,pETB和pETC,在转化大肠杆菌BL21(DE3)中表达,得到了dszA,dszB和dszC基因的融合表达产物.SDS-PAGE分析结果显示,蛋白带分子量分别为50,40和45kDa.3种重组菌BL21(DE3)(pETA),BL21(DE3)(pETB)和BL21(DE3)(pETC)的无细胞粗提液(2mL)的活性检测结果表明,DszC酶的粗提液反应30min能够将0.02mmol/L二苯并噻吩完全转化为二苯并噻吩砜,DszA酶的粗提液能够将0.01mmol/L二苯并噻吩砜完全转化为羟基联苯亚磺酸盐,DszA和DszB酶的粗提液联合作用能够将0.01mmol/L二苯并噻吩砜完全转化为2-羟基联苯,证明LY822对二苯并噻吩的降解符合专一性脱硫的"4S途径"
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Last time updated on 09/11/2016