http://scholars.ntou.edu.tw/handle/123456789/9322
Title: | Molecular Characterization of Mungbean (Vigna radiata L.) Starch Branching Enzyme I | Authors: | Jia-Wei Chang Sing-Chung Li Yun-Chi Shih Reuben Wang Pei-Shan Chung Yuan-Tih Ko |
Keywords: | mungbean;starch branching enzyme;gene expression;recombinant enzyme | Issue Date: | Oct-2010 | Publisher: | ACS Publications | Journal Volume: | 58 | Journal Issue: | 19 | Start page/Pages: | 10437–10444 | Source: | Journal of Agricultural and Food Chemistry | Abstract: | Mungbean (Vigna radiata L. cv. Tainan no. 5) starch branching enzyme I (SBE, EC 2.4.1.18) cDNA, VrsbeI, was cloned, and its expression was characterized. Conserved regions of the family B SBE were used to amplify a full length cDNA of 2208 bp. Phylogeny was analyzed, and the partial 3D structure and functional features were predicted. Catalytic residues were identified in the (α/β)8-fold, and a unique loop from F365 to F376 between β3/α3 was located. Gene expression of VrsbeI in seeds during growth showed that the transcript appeared from week 1 and increased substantially at week 3−4. It was cloned into the pET30 vector and expressed in E. coli BL21(DE3) pLysS cells as a soluble recombinant protein. The affinity-purified recombinant VrSBEI exhibited a specific activity of 314.6 U/mg as an active enzyme with 114-fold activity enrichment from the crude extract. |
URI: | http://scholars.ntou.edu.tw/handle/123456789/9322 | ISSN: | 0021-8561 | DOI: | 10.1021/jf102129f |
Appears in Collections: | 食品科學系 |
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