Effect of pfkA chromosomal interruption on growth, sporulation, and production of organic acids in Bacillus subtilis

• Published in: Journal of basic microbiology 2010-06, Vol.50 (3), p.232-240
• Main Authors: Muñoz-Márquez, María-Enriqueta, Ponce-Rivas, Elizabeth
• Format: Article
• Language: English
• Subjects: Acetic acid; Bacillus subtilis; Bacillus subtilis - enzymology; Bacillus subtilis - genetics; Bacillus subtilis - growth & development; Bacillus subtilis - metabolism; Bacterial Proteins - genetics; Base Sequence; Carbohydrate metabolism; Carbon metabolism; Carboxylic acid; Carboxylic Acids - metabolism; Culture Media - chemistry; Enzymes; Gene Knockout Techniques; Glycolysis; HPLC; Lactic acid; Metabolic Networks and Pathways - genetics; Microorganisms; Models, Biological; Molecular Sequence Data; Mutagenesis, Insertional; Operon; Operons; organic acids; pfkA gene; phosphofructokinase; Phosphofructokinase mutant; Phosphofructokinases - deficiency; Phosphofructokinases - genetics; PTS; pykA gene; Pyruvate kinase; Pyruvate Kinase - genetics; Pyruvate Kinase - metabolism; Pyruvic acid; Spores, Bacterial - growth & development; Sporulation
• ISSN: 0233-111X; 1521-4028
• DOI: 10.1002/jobm.200900236

Phosphofructokinase (Pfk) plays a key role in the regulation of carbohydrate metabolism. Its activity can be used as an indicator of glycolytic flux in a microorganism. We have cloned and characterized the pfkA gene from Bacillus subtilis, which encodes the enzyme phosphofructokinase. This gene was insertionally inactivated at the chromosomal level in a wild type strain and in strains lacking the PEP:sugar phosphotranferase system (PTS). Although the pykA gene is immediately downstream of the pfkA gene, forming a constitutive operon in B. subtilis, the pyruvate kinase activity was not altered in the pfkA mutant. The inactivation of the pfkA gene had a strong impact on the growth of the B. subtilis wild type strain and PTS mutants in Spizizen's minimal media and Schaeffer's sporulation media. Pfk inactivation was also reflected by the timing and percentage of sporulation of the wild type and PTS mutants in sporulation media as well as in the production of organic by-products (pyruvate, lactate, and acetate). (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)