Biotransformation ofR-2-hydroxy-4-phenylbutyric acid byD-lactate dehydrogenase andCandida boidinii cells containing formate dehydrogenase coimmobilized in a fibrous bed bioreactor

Abstract

R-2-hydroxy-4-phenylbutyric acid (R-HPBA) is an important intermediate in the manufacture of angiotensin converting enzyme inhibitors. In this work, a recombinant D-lactate dehydrogenase (LDH) was used to transform 2-oxo-4-phenylbutyric acid (OPBA) to R-HPBA, with concomitant oxidation of beta-nicotinamide adenine dinucleotide (NADH) to NAD(+). The cofactor NADH was regenerated by formate dehydrogenase (FDH) present in whole cells of Candida boidinii, which were pre-treated with toluene to make them permeable. The whole cells used in the process were more stable and easier to prepare as compared with the isolated FDH from the cells. Kinetic study showed that the reaction rate was dependent on the concentration of cofactor, NAD(+), and that both R-HPBA and OPBA inhibited the reaction. A novel method for co-immobilization of whole cells and LDH enzyme on cotton cloth was developed using polyethyleneimine (PEI), which induced the formation of PEI-enzyme-cell aggregates and their adsorption onto cotton cloth, leading to multilayer co-immobilization of cells and enzyme with high loading (0.5 g cell and 8 mg LDH per gram of cotton cloth) and activity yield ( > 95%). A fibrous bed bioreactor with co-immobilized cells and enzyme on the cotton cloth was then evaluated for R-HPBA production in fed-batch and repeated batch modes, which gave relatively stable reactor productivity of 9 g/L . h and product yield of 0.95 mol/mol OPBA when the concentrations of OPBA and R-HPBA were less than 10 g/L.

Bioreductive preparation of ACE inhibitors precursor (R)-2-hydroxy-4-phenylbutanoate esters: Recent advances and future perspectives

“…Compared with classic chemical processes, the asymmetric reduction of OPBE into (R)-HPBE, or the reduction of 2-oxo-4-phneylbutyrate (OPB) to (R)-2-hydroxy-4-phenylbutyrate [(R)-HPB] employing reductases and dehydrogenases ( Figure 2) has several advantageous attributes, including high selectivity, high efficiency, mild reaction condition, and environmental friendless [24][25][26][27]. The complicated and chiral physiological structures endue the enzymes with relatively high chemo-and stereoselectivity and specificity.…”

Enhanced propionic acid production from Jerusalem artichoke hydrolysate by immobilized Propionibacterium acidipropionici in a fibrous-bed bioreactor

“…The fibrous-bed bioreactor (FBB), with a high cell density immobilized in a fibrous matrix packed in the reactor, has been studied in the laboratory during the past 15 years and is a novel cell immobilization process involving both adsorption and entrapment [3,13,14]. It has been successfully used for production of a series of chemicals, including organic acids [15][16][17], enzymes [18,19], xanthan gum [20] and pharmaceuticals, such as developmental endothelial locus-1 [21], embryonic stem cells [22], granulocyte-macrophage colony-stimulating factor (GM-CSF) [23], hepatocytes [24], monoclonal antibody [25] and soluble human Fas ligand [26].…”

Control and Optimization of Clostridium tyrobutyricum ATCC 25755 Adhesion into Fibrous Matrix in a Fibrous Bed Bioreactor

“…By circulating cell suspension through the FBB, a simple in situ immobilization of the cells can be carried out. The FBB has been successfully used for the bioproduction of a series of chemicals, including organic acids [5][6][7], enzymes [8,9], xanthan gum [10], and pharmaceuticals, such as, developmental endothelial locus-1 [11], embryonic stem cells [12], GM-CSF [13], hepatocytes [14], monoclonal antibody [15], and soluble human Fas ligand [16].…”