Sunday, July 29, 2007
P115

Cloning and characterization of S-adenosylsynthetase gene from Pichia ciferrii

SangYoung Yoon, WonKyu Lee*, and YeonWoo Ryu*. Dept of Molecular Science and Technology, Ajou University, palldall hall #427, Ajou University, San 5, Wonchon-dong, Yeongtong-gu ,Suwon ,Korea, Suwon, 443-749, South Korea

S-Adenosylmetionine (SAM) plays a pivotal function in the primary and secondary metabolism of cells as methyl donor in proteins, DNA, RNA, phospholipid, polysaccharide and numerous other molecules. The effect of SAM overexpression from P. ciferrii will be investigated to increase secondary metabolite production because P. ciferrii produce secondary metabolite such as Tetraacetylphytosphingosine (TAPS) as known as precursor of ceramide. However, overall mechanism of SAM synthetase in P. ciferrii is unknown yet. Thus, characterization of SAM synthetase of P. ceferrii has to be investigated prior to following studies.In this research, we isolated SAM synthetase gene from chromosome of P. ciferrii by low-strigency PCR, DNA Walking and RACE to characterize SAM in P. ciferrii. The open reading frame (ORF) of SAM synthetase isolated from P. ciferrii was 1152bp and had a low G+C content of 32.12%. It is encoding a protein of 383 amino acids with 41.8KDa. The highest similarity was to hypothetical protein SAM synthetase 2 gene of its close relative, Debaryomyces hansenii, with 88% similarity. Cloned SAM synthetase gene includes common consensus ATP-binding motif Gly-X-Gly-X-X-Gly and metal binding site (23D and 279D for Mg2+ and 51G for K+). In the recent research, SAM’s effect of morphological differentiation has been reported in several species. Thus, we are in progress of confirmation of morphological differentiation and effect on secondary metabolism production in P. ciferrii.

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