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Applied and Environmental Microbiology, June 2007, p. 3581-3586, Vol. 73, No. 11
0099-2240/07/$08.00+0     doi:10.1128/AEM.02863-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of a Glucokinase with Broad Hexose Specificity from Bacillus sphaericus Strain C3-41{triangledown}

Bei Han,1,2 Haizhou Liu,1,2 Xiaomin Hu,1 Yajun Cai,1,2 Dasheng Zheng,1 and Zhiming Yuan1*

State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China,1 Graduate School of the Chinese Academy of Sciences, Beijing 100039, China2

Received 10 December 2006/ Accepted 17 March 2007

Bacillus sphaericus cannot metabolize sugar since it lacks several of the enzymes necessary for glycolysis. Our results confirmed the presence of a glucokinase-encoding gene, glcK, and a phosphofructokinase-encoding gene, pfk, on the bacterial chromosome and expression of glucokinase during vegetative growth of B. sphaericus strains. However, no phosphoglucose isomerase gene (pgi) or phosphoglucose isomerase enzyme activity was detected in these strains. Furthermore, one glcK open reading frame was cloned from B. sphaericus strain C3-41 and then expressed in Escherichia coli. Biochemical analysis revealed that this gene encoded a protein with a molecular mass of 33 kDa and that the purified recombinant glucokinase had Km values of 0.52 and 0.31 mM for ATP and glucose, respectively. It has been proved that this ATP-dependent glucokinase can also phosphorylate fructose and mannose, and sequence alignment of the glcK gene indicated that it belongs to the ROK protein family. It is postulated that the absence of the phosphoglucose isomerase-encoding gene pgi in B. sphaericus might be one of the reasons for the inability of this bacterium to metabolize carbohydrates. Our findings provide additional data that further elucidate the specific metabolic pathway and could be used for genetic improvement of B. sphaericus.

* Corresponding author. Mailing address: Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. Phone: 86-27-87197242. Fax: 86-27-87198120. E-mail: yzm{at}pentium.whiov.ac.cn

{triangledown} Published ahead of print on 30 March 2007.

Applied and Environmental Microbiology, June 2007, p. 3581-3586, Vol. 73, No. 11
0099-2240/07/$08.00+0     doi:10.1128/AEM.02863-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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