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Journal of Biochemistry Advance Access published online on June 8, 2009
Journal of Biochemistry, doi:10.1093/jb/mvp084
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© The authors 2009. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

Cloning, Expression and Characterization of UDP-Glucose Pyrophosphorylase from Shoots of Bambusa oldhamii

Chia-Jui Weng1,*, Jen-Ying Deng2, Da-Gin Lin3 and Chii-Ling Jeang2

1Graduate Institute of Applied Science of Living, Tainan University of Technology, 529 Jhongjheng Road, Yongkang, Tainan 71002, Taiwan
2Department of Food Science and Biotechnology, National Chung Hsing University, 250 Kuokuang Road, Taichung 40227, Taiwan
3Department of Biotechnology, Transworld Institute of Technology, 1221 Jen-Nang Road, Chia-Tong Li, Douliou, Yunlin 640, Taiwan

*To whom correspondence should be addressed: Dr. Weng, Chia-Jui: Tel: 886-6-2532106 ext.5129, Fax: 886-6-2433837, E-mail: t10044{at}mail.tut.edu.tw

Received May 1, 2009; Accepted May 24, 2009


  Abstract

Carbohydrate biosynthesis for growing cell walls (wall extensibility) is presumed a crucial process to a rapidly growing plant. UDP-glucose pyrophosphorylase (UGPase) is a key enzyme involved in the cell wall synthesis. An UGPase cDNA was isolated from shoots of green bamboo (Bambusa oldhamii), which is a rapid-growing plant and valuable vegetable, by cDNA library screening. The complete sequence of the UGP cDNA was 1744 bp, contained a single open reading frame (ORF) of 1422 nucleotides which is translatable to a polypeptide composed of 473 amino acid residues including a putative transit peptide of 26 amino acid residues. The cloned UGPase of green bamboo shoot is closely related to that of grass family Poaceae of monocot plant by sequence analysis. The recombinant UGPase (rUGPase), which expressed in E. coli system, exhibited higher Km values for G1P and UTP than those of UGPase from bamboo. The reaction mechanism of rUGPase on substrate was shown a compulsory bi-bi mechanism. At various development stages, the UGPase in top portion of bamboo shoots exhibited the highest protein levels and activities than those in other portions. At least four different forms of UDP-glucose-synthesis catalyzing enzymes were existed in bamboo shoot.

Key Words: Bamboo (Bambusa oldhamii), cDNA cloning, Gene expression, UDP-glucose, UDP-glucose pyrophosphorylase


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