Details

Autor(en) / Beteiligte

• Xu, Xiong-Feng
• Hu, Die
• Hu, Bo-Chun
• Li, Chuang
• Liu, You-Yi
• Wu, Min-Chen

Titel

Near-perfect kinetic resolution of o-methylphenyl glycidyl ether by RpEH, a novel epoxide hydrolase from Rhodotorula paludigena JNU001 with high stereoselectivity

Ist Teil von

• Applied microbiology and biotechnology, 2020-07, Vol.104 (14), p.6199-6210

Ort / Verlag

Berlin/Heidelberg: Springer Berlin Heidelberg

Erscheinungsjahr

2020

Quelle

MEDLINE

Beschreibungen/Notizen

• In order to provide more alternative epoxide hydrolases for industrial production, a novel cDNA gene Rpeh -encoding epoxide hydrolase ( Rp EH) 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 Rp EH of 411 amino acids and was heterologously expressed in Escherichia coli BL21(DE3). The substrate spectrum of expressed Rp EH 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, α R and β S , 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 ( ee s ) and 46.7% yield and producing ( S )- 8b with 93.2% ee p 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.