title: Efficient Enantioselective Biocatalytic Production of a Chiral Intermediate of Sitagliptin by a Newly Filamentous Fungus Isolate.
abstract: (S)-3-Hydroxy-1-(3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)-4-(2,4,5-trifluorophenyl)butan-1-one ((S)-HTPP) is a crucial intermediate for the synthesis of Sitagliptin. A fungal strain ZJPH1308, capable of the biocatalysis of ketoamide 4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-one (OTPP) to (S)-HTPP with excellent enantioselectivity, was isolated from a soil sample and identified as Rhizopus microsporus var. rhizopodiformis ZJPH1308 based on its morphological characteristics and internal transcribed spacer (ITS) sequence. Some key reaction parameters involved in the bioreduction catalyzed by isolate ZJPH1308 were then optimized. It demonstrated that the bioreduction of OTPP was effective conducted at relative high temperature (45 °C), along with distilled water as reaction medium and glycerol-coupling approach for cofactor regeneration. Under the optimal conditions, the preparative-scale bioreduction gave a 93.2 % yield (with >99.9 % of enantiomeric excess (ee)) at 15 mM of OTPP and 45 °C, reaction for 24 h. The results indicated that fungal isolate ZJPH1308 can afford a thermostable carbonyl reductase and is a promising biocatalyst for clean and efficient production of valuable chiral intermediate.
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310032, People's Republic of China.
authors.forename: Jia
authors.initials: J
authors.lastname: Sun
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310032, People's Republic of China.
authors.forename: Jin
authors.initials: J
authors.lastname: Huang
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310032, People's Republic of China.
authors.forename: Xuzhong
authors.initials: X
authors.lastname: Ding
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310032, People's Republic of China. wangpu@zjut.edu.cn.
authors.forename: Pu
authors.initials: P
authors.lastname: Wang
mesh_terms: D055162:Biocatalysis / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D005658:Fungi / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism*; D005990:Glycerol / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D010802:Phylogeny; D011719:Pyrazines / Q000378:metabolism; D000068900:Sitagliptin Phosphate / Q000737:chemistry* / Q000378:metabolism*; D013237:Stereoisomerism; D013696:Temperature
chemical_list: D011719:Pyrazines; D005990:Glycerol; D000068900:Sitagliptin Phosphate
keywords: Bioreduction; Distilled water medium; Rhizopus microsporus var. rhizopodiformis; Sitagliptin; Thermophilic carbonyl reductase