Expression and Enzyme Activity Detection of a Sepiapterin Reductase Gene from Musca domestica Larva
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- 作者：Yan Tang ; Zhihua Pei ; Lei Liu ; Dongfang Wang…
- 关键词：Sepiapterin reductase ; Musca domestica ; Gene cloning ; Protein expression ; Enzyme activity
- 刊名：Applied Biochemistry and Biotechnology
- 出版年：2017
- 出版时间：February 2017
- 年：2017
- 卷：181
- 期：2
- 页码：604-612
- 全文大小：
- 刊物类别：Chemistry and Materials Science
- 刊物主题：Biotechnology; Biochemistry, general;
- 出版者：Springer US
- ISSN：1559-0291
- 卷排序：181
文摘
Tetrahydrobiopterin (BH4) is an essential cofactor for aromatic acid hydroxylases and nitric oxide synthase. Sepiapterin reductase (SPR) catalyzes the final steps of BH4 biosynthesis. Studies on SPR from several insects and other organisms have been reported. However, thus far, enzyme activity of SPR in Musca domestica is kept unknown. In this study, 186 differentially expressed genes including SPR gene from Musca domestica (MDSPR) were screened in subtractive cDNA library. The MDSPR gene was cloned, and the recombinant MDSPI16 protein was expressed as a 51-kDa protein in soluble form. The MDSPR exhibited strong activity to the substrate sepiapterin (SP). The values of Vmax and Km of the MDSPR for SP were 6.83 μM/min and 23.48 μM, and the optimum temperature and pH of MDSPR were 50 °C and 4.0, respectively. This study provides new hypotheses and methods for the production of BH4 using insect-derived SPR.