Lactone-ring-cleaving enzyme: genetic analysis, novel RNA editing, and evolutionary implications.
A lactonohydrolase from Fusarium oxysporum AKU 3702 is an enzyme catalyzing the hydrolysis of aldonate lactones to the corresponding aldonic acids. The amino acid sequences of the NH2 terminus and internal peptide fragments of the enzyme were determined to prepare synthetic oligonucleotides as primers for the PCR. An approximate 1, 000-base genomic DNA fragment thus amplified was used as the probe to clone both genomic DNA and cDNA for the enzyme. The lactonohydrolase genomic gene consists of six exons separated by five short introns. A novel type of RNA editing, in which lactonohydrolase mRNA included the insertion of guanosine and cytidine residues, was observed. The predicted amino acid sequence of the cloned lactonohydrolase cDNA showed significant similarity to those of the gluconolactonase from Zymomonas mobilis, and paraoxonases from human and rabbit, forming a unique superfamily consisting of C-O cleaving enzymes and P-O cleaving enzymes. Lactonohydrolase was expressed under the control of the lac promoter in Escherichia coli.
Kobayashi
M
M
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan.
Shinohara
M
M
Sakoh
C
C
Kataoka
M
M
Shimizu
S
S
D000595:Amino Acid Sequence; D000818:Animals; D043303:Aryldialkylphosphatase; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D003562:Cytidine; D004926:Escherichia coli / Q000235:genetics; D004950:Esterases / Q000737:chemistry / Q000235:genetics; D019143:Evolution, Molecular; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics*; D006151:Guanosine; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D017393:RNA Editing; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D017355:Zymomonas / Q000201:enzymology
D012333:RNA, Messenger; D011994:Recombinant Proteins; D006151:Guanosine; D003562:Cytidine; D004950:Esterases; D002265:Carboxylic Ester Hydrolases; C077288:aldonate lactonohydrolase; D043303:Aryldialkylphosphatase