Molecular characterization of mungbean (Vigna radiata L.) starch branching enzyme I

Jia Wei Chang, Sing Chung Li, Yun Chi Shih, Reuben Wang, Pei Shan Chung, Yuan Tih Ko

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4 Citations (Scopus)

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 (R/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.

Original languageEnglish
Pages (from-to)10437-10444
Number of pages8
JournalJournal of Agricultural and Food Chemistry
Volume58
Issue number19
DOIs
Publication statusPublished - Oct 13 2010

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Keywords

  • Gene expression
  • Mungbean
  • Recombinant enzyme
  • Starch branching enzyme

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Chemistry(all)

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