Amination of Ketones by Employing Two New (S)‐Selective ω‐Transaminases and the His‐Tagged ω‐TA from Vibrio fluvialis

Abstract

Two recently identified (S)-selective ω-transaminases (ω-TAs) that originate from Paracoccus denitrificans (Strep-PD-ωTA, cloned with an N-terminal Strep-tag II) and Pseudomonas fluorescens (PF-ωTA) were employed for the asymmetric amination of selected prochiral ketones. The substrates tested were transformed into optically pure amines (Ͼ99 % ee) with high conversion (up to Ͼ99 %). The ω-TAs led to higher conversion in the absence of dimethyl sulfoxide as a cosolvent

A general protein purification and immobilization method on controlled porosity glass: biocatalytic applications

“…The results show that EziG™ is suitable for immobilization of CalA and further optimizations with subsequent applications are ongoing. The FMNH 2 -dependent BVMO 2,5-diketocamphane monooxygenase from Pseudomonas putida (2,5-DKCMO) 32 was coimmobilized on EziG™ with two cofactor-reconverting enzymes (flavin reductase (FRE) from E. coli 33,34 for FMNH 2 recycling and alanine dehydrogenase (AlaDH) from B. subtilis 35 for NADH recycling). This immobilization was performed by exposing the EziG™ carrier to a mixture of all three cell lysates after overexpression of the three enzymes.…”

ω-Transaminases for the amination of functionalised cyclic ketones

“…18,28,46 The nicotinamide cofactor (NADH) required was recycled by employing standard techniques using formate dehydrogenase (FDH) or glucose dehydrogenase (GDH). Two enzyme-coupled systems were used (Scheme 1a), where L-or D-alanine was used as the amine donor and the co-product pyruvate was removed by either a L-lactate dehydrogenase (LDH) (Scheme 1a A) or recycled by an alanine dehydrogenase (AlaDH) for (S)-selective ω-TAms (Scheme 1a B).…”

“…46 (R)-and (S)-8a are commercially available. DNA-modifying enzymes were obtained from Thermo scientific (Germany) or New England Biolabs (USA).…”

3.8.1 Designed Enzymatic Cascades

“…To overcome the task of producing high-yield hydrogen, a new multienzyme system was designed. [137] This system contains five submodules: (i) conversion of cellobiose into glucose 1-phosphate (38) catalyzed by cellobiose phosphorylase (CBP); (ii) generation of glucose 6-phosphate (39) from glucose 1-phosphate (38), catalyzed by phosphoglucomutase (PGM); (iii) production of NADPH, catalyzed by two dehydrogenases (G6PDH and 6GPDH) of the oxidative phase of the pentose phosphate pathway; (iv) regeneration of glucose 6-phosphate (39) from ribulose 5-phosphate (40), catalyzed by the eight enzymes of the non-oxidative phase of the pentose phosphate pathway; and (v) generation of hydrogen from NADPH catalyzed by hydrogenase (H 2 ase) (Scheme 24). Removal of the gaseous products from the aqueous phase at low temperature and pressure is necessary to decrease inhibition, facilitate product separation, and promote the complete reaction.…”