| 纤维微菌海藻糖合成酶基因克隆表达及酶学特性鉴定 |
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| 引用本文： | 曹磊,刘滔滔,张宏涛,谢敏,杜丽琴,梁智群,韦宇拓.纤维微菌海藻糖合成酶基因克隆表达及酶学特性鉴定[J].广西科学,2016,23(1):19-24. |
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| 作者姓名： | 曹磊 刘滔滔 张宏涛 谢敏 杜丽琴 梁智群 韦宇拓 |
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| 作者单位： | 广西大学生命科学与技术学院,广西南宁530005;亚热带农业生物资源保护与利用国家重点实验室,广西南宁 530005 |
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| 基金项目： | 国家自然科学基金项目(31160311)，广西自然科学基金项目(2012GXNSFAA053051) |
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| 摘 要： | 【目的】开发适用于海藻糖生产的新型海藻糖合成酶。【方法】通过反向PCR技术,从一株纤维微菌的基因组DNA中获知海藻糖合成酶基因完整ORF序列,进而克隆得到纤维微菌海藻糖合成酶基因(CCTreS),将其与表达载体pSE380构建重组质粒后转入大肠杆菌BL-21(DE3)中诱导表达,通过镍柱亲和层析纯化得到纯酶并进行酶学性质测定。【结果】从纤维微菌中克隆并在大肠杆菌中成功表达海藻糖合成酶基因(CCTreS)。经纯化获得的重组酶(CCTreS)在以麦芽糖为底物生成海藻糖时,最适反应pH值为7.0,最适反应温度为45℃,40℃保温1h仍具有80%以上的相对酶活力,在pH值5.5~8.5保存24h,相对酶活力仍保留80%以上。Cu~(2+)对其有明显抑制作用。【结论】该重组酶具有很好的热稳定性和pH稳定性,具有一定的研究价值和潜在的工业应用价值。
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| 关 键 词： | 海藻糖合成酶 纤维微菌 反向PCR 酶学性质 |
| 收稿时间： | 2015/11/16 0:00:00 |
| 修稿时间： | 2016/2/26 0:00:00 |
Clonging,Expressionan and Characterization of a Novel Gene Encoding Trehalose Synthase from Cellulosimicrobium cellulans |
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| CAO Lei,LIU Taotao,ZHANG Hongtao,XIE Min,DU Liqin,LIANG Zhiqun and WEI Yutuo.Clonging,Expressionan and Characterization of a Novel Gene Encoding Trehalose Synthase from Cellulosimicrobium cellulans[J].Guangxi Sciences,2016,23(1):19-24. |
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| Authors: | CAO Lei LIU Taotao ZHANG Hongtao XIE Min DU Liqin LIANG Zhiqun and WEI Yutuo |
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| Institution: | College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China and College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China |
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| Abstract: | |
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| Keywords: | trehalose synthase Cellulosimicrobium cellulans inverse-PCR enzymatic characteristics |
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