title: Stabilization of flavobacterium meningosepticum glycerol kinase by introduction of a hydrogen bond.
abstract: The thermostability of Flavobacterium meningosepticum glycerol kinase was increased by the change from Ser329 to Asp [Protein Eng., 14, 663-667 (2001)]. Based on a three-dimensional structure model of the mutant, we have postulated that a new charged-neutral hydrogen bond was formed between Asp329 and Ser414, and the formation of the hydrogen bond contributed to the stabilization of the tertiary structure and increased thermostability of the mutant enzyme. If the postulation is the case, FGK thermostabilization would be possible similarly by the single amino acid substitution from Ser414 to another amino acid which could form the hydrogen bond with Ser329. We did a single amino acid substitution of the wild-type enzyme from Ser414 to Asn. As we expected, S414N showed comparable thermostability to that of S329D. On the other hand, a difference in kinetic properties for ATP between S414N and S329D was observed.
authors.affiliation: Asahi Kasei Corporation, Shizuoka, Japan. sakasegawa.sb@om.asahi-kasei.co.jp
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Sakasegawa
authors.forename: Hideki
authors.initials: H
authors.lastname: Takehara
authors.forename: Issei
authors.initials: I
authors.lastname: Yoshioka
authors.forename: Hideo
authors.initials: H
authors.lastname: Misaki
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Sakuraba
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Ohshima
mesh_terms: D019943:Amino Acid Substitution; D004795:Enzyme Stability; D005417:Flavobacterium / Q000201:enzymology*; D005991:Glycerol Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006860:Hydrogen Bonding; D008958:Models, Molecular; D011487:Protein Conformation; D013696:Temperature
chemical_list: D005991:Glycerol Kinase