Lipases for biocatalysis: development of a chromatographic bioreactor

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 32; no. 4; pp. 715 - 724
• Main Authors: Calleri, E, Temporini, C, Furlanetto, S, Loiodice, F, Fracchiolla, G, Massolini, G
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 08.08.2003; Elsevier Science
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Bioreactors; Candida rugosa; Candida rugosa lipases; Catalysis; Chromatography, High Pressure Liquid - methods; Enantioselective hydrolysis; Fundamental and applied biological sciences. Psychology; Immobilized lipase; Lipase - chemistry; Lipase - metabolism; Immobilized lipase; Candida rugosa lipases; Enantioselective hydrolysis; Enzyme; Triacylglycerol lipase; Esterases; Carboxylic ester hydrolases; Fungi; Non steroidal antiinflammatory agent; Enantiomer; Hydrolases; Fungi Imperfecti; Catalysis; Candida rugosa; Thallophyta
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(03)00179-1

The development of a new chromatographic reactor based on immobilized Candida rugosa lipase (CRL) is described. The chromatographic system has been used to evaluate the rate differences by which the product enantiomers of esterolytic reactions catalyzed by immobilized CRL are obtained. The method has been applied to a series of racemic 2-aryloxyalkanoic acids and isosteric analogous methyl esters and to some non-steroidal antiinflammatory drugs 2-arylpropanoic acids methyl esters in order to study the structure effects on reaction rate and enantioselectivity. Lipase from C. rugosa has been non-covalently and covalently immobilized on HPLC chromatographic silica supports to develop an immobilized enzyme reactor (IMER). The reactor was connected through a switching valve to an analytical reversed-phase column, which was used for the on-line determination of the hydrolysis rate by peak area integration. The enantiomeric excess of the hydrolytic reaction products was determined off-line on a CSP utilizing immobilized penicillin G acylase (PGA-CSP).