(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) > cellular organisms: NE > Eukaryota: NE > Opisthokonta: NE > Fungi: NE > Dikarya: NE > Basidiomycota: NE > Pucciniomycotina: NE > Microbotryomycetes: NE > Sporidiobolales: NE > Sporidiobolaceae: NE > Rhodotorula: NE > Rhodotorula paludigena: NE
LegendThis sequence has been compared to family alignement (MSA) red => minority aminoacid blue => majority aminoacid color intensity => conservation rate title => sequence position(MSA position)aminoacid rate Catalytic site Catalytic site in the MSA MAAHSFTAPPAPYNIDFAPQVDDLHRRLDAARWPGQDVVPDDVDYGEHGA FGLGAGPSLALMKELAQEWRGQDQKQLQDHLNSYKNYRVEIEGLDIHFLH YPSARADAFPLILCHGWPGGYHEFLHVLERLTEPKDQGSRAFHVVVPSMP GYAFSSPPKTAKWGMEDTARVFDKLMTGLGYVKYAAQGGDWGSITARCLG SLHKENCVAVHLNFCPVPPPFPLNMFNPRTLLDWMPRFVLPDERRAKLER GVAYIERGSSYYAMQNLTPRTPAYGLNDSPIGLLAWIGEKMIPGIDKAVK HPNATLNREALFTTLSIYWFTGSIGSSFLPYALNPHFGTFLVSPRHHLPN FALSNFPDELFTPEERDARRTGNLRWYKDAEDGGHFAALEKPEVFAEHVR EAMGVLLSNQA
Reference
Title: Near-perfect kinetic resolution of o-methylphenyl glycidyl ether by RpEH, a novel epoxide hydrolase from Rhodotorula paludigena JNU001 with high stereoselectivity Xu XF, Hu D, Hu BC, Li C, Liu YY, Wu MC Ref: Applied Microbiology & Biotechnology, :, 2020 : PubMed
In order to provide more alternative epoxide hydrolases for industrial production, a novel cDNA gene Rpeh-encoding epoxide hydrolase (RpEH) of Rhodotorula paludigena JNU001 identified by 26S rDNA sequence analysis was amplified by RT-PCR. The open-reading frame (ORF) of Rpeh was 1236 bp encoding RpEH of 411 amino acids and was heterologously expressed in Escherichia coli BL21(DE3). The substrate spectrum of expressed RpEH showed that the transformant E. coli/Rpeh had excellent enantioselectivity to 2a, 3a, and 5a-10a, among which E. coli/Rpeh had the highest activity (2473 U/g wet cells) and wonderful enantioselectivity (E = 101) for 8a, and its regioselectivity coefficients, alphaR and betaS, toward (R)- and (S)-8a were 99.7 and 83.2%, respectively. Using only 10 mg wet cells/mL of E. coli/Rpeh, the near-perfect kinetic resolution of rac-8a at a high concentration (1000 mM) was achieved within 2.5 h, giving (R)-8a with more than 99% enantiomeric excess (ees) and 46.7% yield and producing (S)-8b with 93.2% eep and 51.4% yield with high space-time yield (STY) for (R)-8a and (S)-8b were 30.6 and 37.3 g/L/h.
