Chiral diol t-butyl 6-cyano-(3R,5R)-dihydroxylhexanoate synthesis catalyzed by immobilized cells of carbonyl reductase and glucose dehydrogenase co-expression E. coli

In our previous work, we constructed a carbonyl reductase (CR) and glucose dehydrogenase (GDH) co-expression strain E. coli BL21(DE3)/pCDFDuet- gdh - cr , and a biocatalytic process of chiral diol t -butyl 6-cyano-(3 R ,5 R )-dihydroxyhexanoate ((3 R ,5 R )- 2 ) was established by using its free cells. Herein, a (3 R ,5 R )- 2 process with celite-polyethyleneimine (PEI)-glutaraldehyde (GA) immobilized E. coli BL21(DE3)/pCDFDuet- gdh - cr whole cells was developed for the first time. It was demonstrated that immobilization via cross-linking was preferred, and the optimized immobilization conditions were as follows: 0.6% (w/v) celite, 4% (w/v) PEI and 1% (w/v) GA, pH7. The immobilizate can be used at pH 7.0, 30   °C for intermittent 7 batch, yielding a total of 17.5   g (3 R ,5 R )- 2 per gram of celite-PEI-GAimmobilizates in de p (diastereomeric excess) >99.5%. Compared with the free cells, there was a dramatic improvement in celite- PEI-GA immobilizate stability and recyclability. In this way, a promising platform technology based on a combination of cells immobilization and multiple enzymes co-expression is constructed, enabling efficiently converting inexpensive prochiral ketones to high-valued chiral alcohols.