Kinetic modelling of coupled redox enzymatic systems for in situ regeneration of NADPH

Kinetic aspects involved in the simultaneous enzymatic synthesis of (S)-3-fluoroalanine and (R)-3-fluorolactic acid

“…However, the use of dehydrogenases in preparative applications requires effective processes for coenzyme regeneration since these cofactors are too expensive to be used as stoichiometric reagents. Several of these processes have already been reported by our group [4][5][6].…”

“…Since the kinetic properties of L-ALADH acting on rac-3-fluoroalanine as well as those of L-LDH catalyzing the enantioselective reduction of 3-fluoropyruvate [5,6] were studied by our group, we envisaged an enzymatic system suitable for the simultaneous production of (S)-3-fluoroalanine (1a) and (R)-3-fluorolactic acid (3). As shown in Scheme 3 this enzymatic system involves the deaminative oxidation of the R-enantiomer (1b) of rac-1 to produce NH 4 þ , 3-fluoropyruvate (2) and NADH.…”

“…The use of isolated enzymes to catalyze these reactions has greatly increased in the last years [1][2][3][4][5][6][7][8][9][10][11][12].…”

“…(R)-3-Fluorolactic acid was obtained enzymatically in our laboratory in 80% overall yield and ee > 99% in a system involving the enantioselective reduction of 3-fluoropyruvate catalyzed by L-LDH and NADH which was in situ regenerated by the oxidation of cis-1,2-bis(hydroxymethyl)cyclohexane catalyzed by HLADH [5,6]. (R)-3-Fluoroalanine was obtained by Oshima et al [12] with L-ALADH from Bacillus sphaericus by reductive amination of 3-fluoropyruvate in the presence of NADH.…”

New kinetic and thermodynamic approach of one-pot synthesis of chiral alcohol by oxidoreductase from Candida parapsilosis

“…Recent literature [24][25][26] showed that some of these enzymes will follow a special form of ordered BiBi mechanism, the TheorellChance BiBi mechanism, which does not form central ternary complexes. The simplest method, the study of product inhibition caused by the first product of the reduction reaction compared to the second substrate of the carbonyl reductase, was carried out to analyze the kinetic significance of the ternary complexes (Table 3).…”