Genetic and biochemical characterization of the two glutamine synthetases GSI and GSII of the phosphinothricyl-alanyl-alanine producer, T494 Free

Abstract

SUMMARY: The 1410 bp DNA region () encoding glutamine synthetase I (GSI) from was amplified by PCR, cloned and sequenced. The molecular mass of the deduced GSI protein (469 residues) was determined to be 50 kDa. The DNA region showed 90% nucleotide identity with the A3(2) gene, but no significant nucleotide sequence similarity with the (GSII) gene of The chromosomal and genes of were disrupted by site-specific mutagenesis. Neither nor single mutants required glutamine for growth and both were normal in their sporulation. Measurement of the GS activity in cultures grown with different nitrogen sources revealed that GSI (heat-stable) and GSII (heat-labile) were always expressed together, with GSI as the predominant activity. It could be proposed that GSI, but not GSII is inactivated by adenylylation under conditions of nitrogen excess. GSI and GSII activities are inhibited by amino acids and by nucleotides.

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1993-08-01
2024-03-29
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