Molecular cloning and functional expression of D-sorbitol dehydrogenase from Gluconobacter suboxydans IF03255, which requires pyrroloquinoline quinone and hydrophobic protein SldB for activity development in E. coli.
The sldA gene that encodes the D-sorbitol dehydrogenase (SLDH) from Gluconobacter suboxydans IFO 3255 was cloned and sequenced. It encodes a polypeptide of 740 residues, which contains a signal sequence of 24 residues. SLDH had 35-37% identity to the membrane-bound quinoprotein glucose dehydrogenases (GDHs) from E. coli, Gluconobacter oxydans, and Acinetobacter calcoaceticus except the N-terminal hydrophobic region of GDH. Additionally, the sldB gene located just upstream of sldA was found to encode a polypeptide consisting of 126 very hydrophobic residues that is similar in sequence to the one-sixth N-terminal region of the GDH. For the development of the SLDH activity in E. coli, co-expression of the sldA and sldB genes and the presence of pyrrloquinolone quinone as a co-factor were required.
Miyazaki
Taro
T
Department of Applied Microbiology, Nippon Roche Research Center, Kamakura, Kanagawa, Japan.
Tomiyama
Noribumi
N
Shinjoh
Masako
M
Hoshino
Tatsuo
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D020576:Gluconobacter / Q000201:enzymology*; D007064:L-Iditol 2-Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D045542:PQQ Cofactor; D016133:Polymerase Chain Reaction; D015363:Quinolones / Q000378:metabolism*; D011809:Quinones / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D015363:Quinolones; D011809:Quinones; D045542:PQQ Cofactor; D007064:L-Iditol 2-Dehydrogenase