| 耐辐射奇球菌类胡萝卜素C3',4'-脱氢酶底物特异性 |
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| 引用本文： | 孙宗涛,田兵,沈绍传,华跃进.耐辐射奇球菌类胡萝卜素C3',4'-脱氢酶底物特异性[J].生物工程学报,2010,26(10):1451-1455. |
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| 作者姓名： | 孙宗涛 田兵 沈绍传 华跃进 |
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| 作者单位： | 1. 浙江大学原子核农业科学研究所,杭州,310029;浙江省农业科学院病毒学与生物技术所,杭州,310021
2. 浙江大学原子核农业科学研究所,杭州,310029 |
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| 基金项目： | 国家重点基础研究发展计划 (973计划) (No. 2007CB707804)，国家高技术研究发展计划 (863计划) (No. 2007AA021305)，国家自然科学基金项目 (Nos. 30830006, 30670026, 30870035)，农业部核技术农业应用项目 (No. 200803034) 资助。 |
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| 摘 要： | 为了鉴定耐辐射奇球菌类胡萝卜素C3',4'-脱氢酶(DR2250)催化底物的特异性,利用PCR方法将dr2250基因的克隆到载体pUC19上,形成了重组载体pUC-CRTD。利用质粒共转化方法将不同组合的质粒pACCRT-EBIEu、pRK-CRTC和pUC-CRTD转化到大肠杆菌中,筛选阳性克隆并提取其色素进行产物分析。结果表明,DR2250修饰的底物具有选择性,它不能以未经过羟基化修饰的直链类胡萝卜素为底物,而能在C1(1')羟基修饰的羟基化类胡萝卜素的基础上进行C3',4'-脱氢反应。
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| 关 键 词： | 耐辐射奇球菌，类胡萝卜素，C3' 4'-脱氢酶 底物特异性 |
| 收稿时间： | 6/2/2010 12:00:00 AM |
Substrate specificity of carotenoid 3',4'-desaturase from Deinococcus radiodurans |
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| Zongtao Sun,Bing Tian,Shaochuan Shen and Yuejin Hua.Substrate specificity of carotenoid 3'',4''-desaturase from Deinococcus radiodurans[J].Chinese Journal of Biotechnology,2010,26(10):1451-1455. |
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| Authors: | Zongtao Sun Bing Tian Shaochuan Shen and Yuejin Hua |
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| Institution: | Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310029, China; Department of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310029, China;Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310029, China;Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310029, China |
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| Abstract: | To examine the substrate specificity of carotenoid 3',4'-desaturase (DR2250) from Deinococcus radiodurans, we amplified the dr2250 gene by using PCR methods. The PCR products were digested by Hind III-BamH I and ligated into the vector pUC19, yielding recombinant vector pUC-CRTD. We analyzed the carotenoids of E. coli transformants containing pACCRT-EBIEu and (or) pRK-CRTC and (or) pUC-CRTD. Our results demonstrated that DR2250 had substrate specificity on the carotenoids with hydroxyl group at C1 (1'). |
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| Keywords: | Deinococcus radiodurans carotenoid C3' 4'-desaturase substrate specificity |
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