Cloning, sequence analysis, and expression of the gene encoding Sphingomonas paucimobilis FP2001 alpha-L -rhamnosidase.
The gene (rhaM) encoding the alpha-L-rhamnosidase of Sphingomonas paucimobilis FP2001 was cloned, sequenced, and expressed in Escherichia coli. The rhaM consisted of 3,354 nucleotides and had a promoter and Shine-Dalgarno sequences typical in bacteria. The rhaM encoding a protein (Rham) deducted from the sequence consisted of 1,117 amino acids and had a putative signal peptide of 25 amino acids. Rham has no similarity to other known rhamnosidases. Rham has a sugar-binding domain of glycoside hydrolase family 2, which has been well conserved in beta-glucuronidase, beta-mannosidase, and beta-galactosidase, in its C-terminal region. Rham is possibly a member of a new bacterial subfamily in glycoside hydrolase family 78 (alpha-L-rhamnosidase). RT-PCR analysis of rhaM mRNA indicated that the induction of alpha-L-rhamnosidase by the addition of L-rhamnose occurred on the transcriptional level.
Miyata
Takeshi
T
Microbiology Laboratory, Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan-ku, Fukuoka 814-0180, Japan.
Kashige
Nobuhiro
N
Satho
Tomomitsu
T
Yamaguchi
Tadatoshi
T
Aso
Yoichi
Y
Miake
Fumio
F
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017421:Sequence Analysis; D020578:Sphingomonas / Q000201:enzymology* / Q000235:genetics
D006026:Glycoside Hydrolases; C022525:alpha-L-rhamnosidase