TAILIEUCHUNG - Cloning, expression, and characterization of a novel CTP synthase gene from Anoxybacillus gonensis G2

The cytidine-5’-triphosphate (CTP) synthase (EC ) gene (pyrG) was cloned and sequenced from the thermophilic bacterium Anoxybacillus gonensis G2 (Ago). The gene is 1590 bp in length and encodes a protein of 530 amino acids, with a molecular mass of kDa. | Turkish Journal of Biology Turk J Biol (2014) 38: 111-117 © TÜBİTAK doi: Research Article Cloning, expression, and characterization of a novel CTP synthase gene from Anoxybacillus gonensis G2 1, 1,2 1 3 4 5 Cemal SANDALLI *, Ayşegül SARAL , Serdar ÜLKER , Hakan KARAOĞLU , Ali Osman BELDÜZ , Ayşegül ÇOPUR ÇİÇEK 1 Department of Biology, Faculty of Arts and Sciences, Recep Tayyip Erdoğan University, Rize, Turkey 2 Department of Biology, Faculty of Arts and Sciences, Artvin Çoruh University, Artvin, Turkey 3 Department of Aquaculture, Faculty of Fisheries, Recep Tayyip Erdoğan University, Rize, Turkey 4 Department of Biology, Faculty of Sciences, Karadeniz Technical University, Trabzon, Turkey 5 Department of Medical Microbiology, Faculty of Medicine, Recep Tayyip Erdoğan University, Rize, Turkey Received: Accepted: Published Online: Printed: Abstract: The cytidine-5’-triphosphate (CTP) synthase (EC ) gene (pyrG) was cloned and sequenced from the thermophilic bacterium Anoxybacillus gonensis G2 (Ago). The gene is 1590 bp in length and encodes a protein of 530 amino acids, with a molecular mass of kDa. The amino acid sequence of CTP synthase shares approximately 90%–94% similarity to Bacillus sp., and it belongs to the triad glutamine amidotransferases, which utilize a Cys–His–Glu triad for activity. Multiple sequence alignments revealed that the enzyme includes conserved amino acids responsible for catalytic activity and the binding of a divalent metal ion (Mg+2). AgoCTP synthase (AgoG2CTPs) was overproduced in Escherichia coli BL21 (DE3) pLysS as recombinant and purified by nickel affinity chromatography. Its biochemical characterization showed that the enzyme had maximal activity at pH – and 65 °C. Km, Vmax, and kcat were found to be approximately mM, U/L, and s–1 at 65 °C, respectively. CTP synthase promotes the .

• Sinh học
• Anoxybacillus gonensis
• Cytidine 5’ triphosphate synthase
• NH3 dependent characterization
• Amino acids
• Uridine monophosphate
• Uridine triphosphate
• Xylose isomerase
• Anoxybacillus gonensis G2T
• E
• coli
• Taq DNA Polymerase
• Unincubated enzyme