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

Research output: Contribution to journalArticle

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

Fingerprint

1,4-alpha-Glucan Branching Enzyme
1,4-alpha-glucan branching enzyme
Vigna radiata
mung beans
Complementary DNA
Phylogeny
Complex Mixtures
Recombinant Proteins
recombinant proteins
Gene expression
Escherichia coli
Seed
Seeds
Gene Expression
gene expression
phylogeny
extracts
Enzymes
Growth
enzymes

Keywords

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

ASJC Scopus subject areas

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

Cite this

Molecular characterization of mungbean (Vigna radiata L.) starch branching enzyme I. / Chang, Jia Wei; Li, Sing Chung; Shih, Yun Chi; Wang, Reuben; Chung, Pei Shan; Ko, Yuan Tih.

In: Journal of Agricultural and Food Chemistry, Vol. 58, No. 19, 13.10.2010, p. 10437-10444.

Research output: Contribution to journalArticle

Chang, Jia Wei ; Li, Sing Chung ; Shih, Yun Chi ; Wang, Reuben ; Chung, Pei Shan ; Ko, Yuan Tih. / Molecular characterization of mungbean (Vigna radiata L.) starch branching enzyme I. In: Journal of Agricultural and Food Chemistry. 2010 ; Vol. 58, No. 19. pp. 10437-10444.
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