To develop novel antimicrobial agents to Rhizoctonia solani, the genomic gene and complete cDNA encoding glucosamine6phosphate synthetase were cloned and sequenced from the rice pathogen Rhizoctonia solani by 3′RACE and 5′RACE . Homologous to other reported GlmS in sequence, the GlmS contains eight introns, and encodes a predicted protein of 697 amino acids. Domain structure analysis revealed that R.solani GlmS contained a glutamine transferase motif and two sugar isomerase motifs. Recombinant native R.solani GlmS enzyme was overexpressed using Escherichia coli and purified. The results of Gel filtration chromatography and SDSPAGE revealed that it had an estimated molecular mass of 306 kD and consisted of four equalsized subunits of 77 kD. The optimal reaction conditions for the recombinant GlmS were pH 6.4 at 37℃, the halflife period for the recombinant GlmS was 1 h at 42℃ and the enzyme was stable at pH 5.5-7.5. R.solani GlmS activity was inhibited by the endproduct of the hexosamine pathway, UDPGlcNAc.
LUO Chuping1,3, LIU Yongfeng1, CHEN Zhiyi1,3，*, WANG Xiaoyu1, FANG Xianwen2, CHEN Zhongming2, LIU Youzhou1, NIE Yafeng1 , ZHANG Rongsheng1
. Molecular Cloning, Sequencing, and Expression of a Glucosamine6phosphate Synthetase Gene from Rice Pathogen Rhizoctonia solani [J]. Chinese Journal OF Rice Science, 2012
, 26(2)
: 137
-143
.
DOI: 10.3969/j.issn.10017216.2012.02.002
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