원문정보
초록
영어
About 80% of chiral amines used as pharmaceutical intermediates are in (R) form. Despite the needs of (R) form intermediates, only one (R)-specific ω-transaminase (R-ωAT) is known. In this study, we used bioinformatical approach to search putative R-ωAT. Because R-ωAT is believed to belong to aminotransferase group III which contains D-alanine aminotransferase, branched-chain aminotransferase and 4-aminodeoxychorismate lyase (ADCL), we generated profiles of DAATs, BCATs and ADCLs and scored bacterial sequences in RefSeq database using three profiles. We plotted sequences in 3-D space whose coordinates are composed of profile analysis score against DAATs, BCATs and ADCLs. Slightly homologous to DAAT, BCAT and ADCL, but not authentic DAAT, BCAT nor ADCL, were selected for activity investigation. Reaction results using enzyme purified from recombinant E. coli showed that transaminases from Hypomonas neptunium and Saccharopolyspora erythrea showed activity to (R)-a-MBA. These two enzymes did not showed activity to (S)-a-MBA. Therefore, R-ωAT were successfully screened using evolutionary relationship by comparison of profile analysis scores against neighborhood subfamily profiles.