Cloning, expression, and characterization of acetate kinase from Lactobacillus sanfranciscensis.
In the metabolism of Lactobacillus sanfranciscensis, the acetate kinase (AK) is a key enzyme and responsible for dephosphorylation of acetyl phosphate with the concomitant production of acetate and ATP. The L. sanfranciscensis ack gene was identified by PCR methods. It encodes a 397 amino acid protein sharing 56% similarity with Bacillus subtilis AK. Whereas cotranscription of ack and pta (phosphotransacetylase) is reported in previously characterised organisms, the L. sanfranciscensis ack gene is not located in direct neighbourhood to the encoding gene. AK was heterologously expressed in E. coli and characterised by its v(max) and Km values and by the dependence of enzyme activity on temperature and pH. Based on this data the in vivo role of the enzyme is discussed.
Knorr
R
R
Institut für Technische Mikrobiologie, Technische Universität München, Freising-Weihenstephan, Germany.
Ehrmann
M A
MA
Vogel
R F
RF
D000084:Acetate Kinase / Q000235:genetics* / Q000378:metabolism; D000085:Acetates / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007778:Lactobacillus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010755:Organophosphates / Q000378:metabolism; D010766:Phosphorylation; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D000085:Acetates; D004269:DNA, Bacterial; D010755:Organophosphates; C011632:acetyl phosphate; D000255:Adenosine Triphosphate; D000084:Acetate Kinase