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Wei Sheng Wu Xue Bao. 2008 May;48(5):602-7.

[Cloning, expression and elementary characterization of phosphofructokinase from Bacillus sphaericus C3-41].

[Article in Chinese]

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Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.



Bacillus sphaericus is unable to use hexose and pentoses as the sole carbon source, due to the lack of key enzymes in Embden-Meyerhof-Parnas pathway (EMP), Hexose Monophophate Pathway (HMP) and Entner-Doudoroff (ED) pathway, such as phosphofructokinase (PFK). Based on the genome sequence annotation results of B. sphaericus C3-41, the phosphofructokinase gene pfk was verified with a single copy on chromosome, the aim of this research is to analysis the EMP pathway in B. sphaericus further, and confirm the function of phosphofructokinase.


The methods of southern-blot of pfk gene among different B. sphaericus strains, pfk ORF cloning from C3-41 and expressing in Escherichia coli, the corresponding sequence analysis and anlignment were used.


The pfk ORF of B. sphaericus was composed of 960 bp nucleitides encoding a protein about 42 kDa, and the PFK sequence analysis showed it had the conservative amino acids-binding sites and an ATP-binding domain. The expression of pfk in recombinant E. coli strain could complement the PFK activity of a pfk mutated E. coli strain DF1020.


The expressed PFK had the conventional phosphofructokinase activity, and settled the foundation for the further research of catabolism of B. sphaericus.

[Indexed for MEDLINE]

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