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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제26권 제6호
- 발행연도
- 2016.1
- 수록면
- 1,077 - 1,086 (10page)
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초록· 키워드
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Glycerol dehydrogenases (GlyDHs) are essential for glycerol metabolism in vivo, catalyzing its reversible reduction to 1,3-dihydroxypropranone (DHA). The gldA gene encoding a putative GlyDH was cloned from Thermoanaerobacterium thermosaccharolyticum DSM 571 (TtGlyDH) and expressed in Escherichia coli. The presence of Mn2+ enhanced its enzymatic activity by 79.5%. Three highly conserved residues (Asp171, His254, and His271) in TtGlyDH were associated with metal ion binding. Based on an investigation of glycerol oxidation and DHA reduction, TtGlyDH showed maximum activity towards glycerol at 60°C and pH 8.0 and towards DHA at 60°C and pH 6.0. DHA reduction was the dominant reaction, with a lower Km(DHA) of 1.08 ± 0.13 mM and Vmax of 0.0053 ± 0.0001 mM/s, compared with glycerol oxidation, with a Km(glycerol) of 30.29 ± 3.42 mM and Vmax of 0.042 ± 0.002 mM/s. TtGlyDH had an apparent activation energy of 312.94 kJ/mol. The recombinant TtGlyDH was thermostable, maintaining 65% of its activity after a 2-h incubation at 60°C. Molecular modeling and site-directed mutagenesis analyses demonstrated that TtGlyDH had an atypical dinucleotide binding motif (GGG motif) and a basic residue Arg43, both related to dinucleotide binding.
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참고문헌 (30)
참고문헌 신청
Altschul SF / 1990 / Basic local alignment search tool / J. Mol. Biol 215:403~410
Arnold K / 2006 / The SWISSMODEL workspace: a web-based environment for protein structure homology modelling / Bioinformatics 22:195~201
Bellamacina CR / 1996 / The nicotinamide dinucleotide binding motif: a comparison of nucleotide binding proteins / FASEB J 10:1257~1269
Bruins ME / 2001 / Thermozymes and their applications: a review of recent literature and patents / Appl. Biochem. Biotechnol 90:155~186
Burke J / 2001 / Purification, crystallization and quaternary structure analysis of a glycerol dehydrogenase S305C mutant from Bacillus stearothermophilus / Acta Crystallogr. D Biol. Crystallogr 57:165~167
Arnold K / 2006 / The SWISSMODEL workspace: a web-based environment for protein structure homology modelling / Bioinformatics 22:195~201
Bellamacina CR / 1996 / The nicotinamide dinucleotide binding motif: a comparison of nucleotide binding proteins / FASEB J 10:1257~1269
Bruins ME / 2001 / Thermozymes and their applications: a review of recent literature and patents / Appl. Biochem. Biotechnol 90:155~186
Burke J / 2001 / Purification, crystallization and quaternary structure analysis of a glycerol dehydrogenase S305C mutant from Bacillus stearothermophilus / Acta Crystallogr. D Biol. Crystallogr 57:165~167
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