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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]

Author information

1
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. tydyhan@gmail.com

Abstract

OBJECTIVE:

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.

METHODS:

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.

RESULTS:

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.

CONCLUSION:

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

PMID:
18652291
[Indexed for MEDLINE]

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