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

doi:10.1021/jf102129f

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

School of Nutrition and Health Sciences

Research output: Contribution to journal › Article › peer-review

5 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 language	English
Pages (from-to)	10437-10444
Number of pages	8
Journal	Journal of Agricultural and Food Chemistry
Volume	58
Issue number	19
DOIs	https://doi.org/10.1021/jf102129f
Publication status	Published - Oct 13 2010

Keywords

Gene expression
Mungbean
Recombinant enzyme
Starch branching enzyme

ASJC Scopus subject areas

Agricultural and Biological Sciences(all)
Chemistry(all)

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10.1021/jf102129f

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title = "Molecular characterization of mungbean (Vigna radiata L.) starch branching enzyme I",

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.",

keywords = "Gene expression, Mungbean, Recombinant enzyme, Starch branching enzyme",

author = "Chang, {Jia Wei} and Li, {Sing Chung} and Shih, {Yun Chi} and Reuben Wang and Chung, {Pei Shan} and Ko, {Yuan Tih}",

year = "2010",

month = oct,

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doi = "10.1021/jf102129f",

language = "English",

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T1 - Molecular characterization of mungbean (Vigna radiata L.) starch branching enzyme I

AU - Chang, Jia Wei

AU - Li, Sing Chung

AU - Shih, Yun Chi

AU - Wang, Reuben

AU - Chung, Pei Shan

AU - Ko, Yuan Tih

PY - 2010/10/13

Y1 - 2010/10/13

N2 - 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.

AB - 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.

KW - Gene expression

KW - Mungbean

KW - Recombinant enzyme

KW - Starch branching enzyme

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

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Keywords

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