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Troshina, O, Hansel, A and Lindblad, P (2001) Cloning, characterization, and functional expression in Escherichia coli of argH encoding argininosuccinate lyase in the cyanobacterium Nostoc sp. strain PCC 73102. Curr. Microbiol. 43:260-4


A gene argH, encoding argininosuccinate lyase (ASL), has been cloned from a cosmid library of the filamentous cyanobacterium Nostoc sp. strain PCC 73102. The argH open reading frame encodes a protein comprised of 461 amino acids with a calculated molecular mass of 51,349 Da. Protein sequence comparisons reveal significant similarities of the Nostoc PCC 73102 ASL to related proteins from other organisms. In an Escherichia coli delta argH strain, the Nostoc PCC 73102 ASL expressed from a recombinant plasmid could restore the ability to grow on medium without arginine. Moreover, cell extracts show a specific ASL activity of 16.2 nmoles of urea x min(-1) x (mg protein)(-1). Partially purified, His-tagged ASL runs as a 53-kDa protein band in SDS-PAGE and about 215-kDa protein in native-PAGE, suggesting that the native protein is a tetramer.




Amino Acid Sequence; Argininosuccinate Lyase/chemistry; Argininosuccinate Lyase/genetics; Argininosuccinate Lyase/metabolism; Cloning, Molecular; Cyanobacteria/enzymology; Cyanobacteria/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/growth & development; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Genetic Complementation Test; Molecular Sequence Data



Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status


GO:0004056: argininosuccinate lyase activity



E. coli C600 (WT functional argH),E. coli W3678 (no argH gene), C. jejuni TGH9011 (WT functional argH), and transformed E. coli (C. jejuni TGH9011 argH gene) were examined for ASL activity. E. coli C600, which is wild type with respect to the argH gene, showed a specific activity of 1.1 + 0.17 ,umol of arginine per h per mg of protein. C. jejuni TGH9011 showed a specific activity of 1.3 ± 0.32 ,umol of arginine per h per mg of protein. E. coli W3678 did not possess any significant ASL activity compared with extract-free controls. The transformed strain pARGH1-1 had a specific activity of 5.7 ± 1.2 ,imol of arginine per h per mg of protein. This result confirms that the protein derived from the recombinant plasmid possesses argininosuccinate lyase activity activity.

CACAO 4555

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