Enantioselective Hydrolysis of a meso-Diester Using Pig Liver Esterase in a Two-Phase Stirred Tank Reactor

Abstract

The enzymatic hydrolysis of the meso-diester cis-cyclohex-4-ene-1,2-dicarboxylate to the monoester methyl-hydrogen(1S,2R)-cyclohex-4-ene-1,2-dicarboxylate using pig liver esterase has been studied both experimentally and theoretically. A chiral monoester product of high pharmaceutical interest is obtained with an enantiomeric excess of 97%. The enzymatic conversion of the meso-diester compound highly soluble in organic solvents to the desired monoester highly soluble in aqueous phase was studied in a two-phase contactor under controlled hydrodynamic conditions and interfacial contact area. A comprehensive mathematical model comprising mass transfer and simultaneous enzymatic conversion was developed. This model proved to be a simple and useful tool, making it possible to simulate the process under different conditions and to optimize the most relevant operating parameters, without resorting to exhaustive laboratory studies.

Scale-Up of a Recombinant Pig Liver Esterase-Catalyzed Desymmetrization of Dimethyl Cyclohex-4-ene-cis-1,2-dicarboxylate

“…The other enantiomer, (1 S ,2 R) -monoester 2a , was repeatedly synthesized via a pig liver esterase-catalyzed reaction. Unfortunately the reaction is not entirely enantioselective, and a lower enantiomeric excess of 80–97% ee was obtained (Scheme ). ,− The reduced enantioselectivity results from the presence of several parallel hydrolytic reactions, each independently catalyzed by different isoenzymes of pig liver esterase. The products of highly enantioselective isoenzymes are hereby mixed with products from less selective enzymes.…”

Chemoenzymatic One‐Pot Synthesis of Enantiopure L‐Arylalanines from Arylaldehydes

“…Enzymes of broad substrate specificity, while maintaining other specificities, are therefore preferred as biocatalysts. Porcine liver esterase (PLE) [4][5][6][7] and phenylalanine ammonia lyase (PAL) [8] fulfil these criteria.…”

Enzymatic Desymmetrization of Dimethyl Cylcohex-4-ene-cis-1,2-dicarboxylate to (1S,2R)-2-(Methoxycarbonyl)cyclohex-4-ene-1-carboxylic Acid

“…The enzymes could be classified in three groups (Table ): Entries 1 and 2: Porcine liver esterase , and acylase I from porcine liver showed high (99%) and good (61%) conversions, respectively, to the undesired 1 R ,2 S -monoester 2 in high (94−95%) ee. Entries 3 to 6: Novozym 435, lipoprotein lipase, and cholesterol esterase showed high conversion (93−100%) to the desired 1 S ,2 R -monoester 1 with high (86−99%) ee. Lipase TL showed good (51%) conversion with moderate (39%) ee to the desired 1 S ,2 R -monoester 1 . Entries 7 to 11: These enzymes showed low conversions (6−15%) in moderate to high (45−82%) ee to the desired 1 S ,2 R -monoester ( 1 ). …”

“…Entries 1 and 2: Porcine liver esterase , and acylase I from porcine liver showed high (99%) and good (61%) conversions, respectively, to the undesired 1 R ,2 S -monoester 2 in high (94−95%) ee.…”

“…There are several reports in the literature , for the synthesis of the opposite enantiomer of the monoester, (1 R ,2 S )-2-(methoxycarbonyl)cyclohex-4-ene-1-carboxylic acid, 2 , by porcine liver enzyme-catalyzed hydrolytic desymmetrization of the dimethyl ester 4 . We hypothesized that a broad screen of enzymes would identify an enzyme that would similarly afford the desired enantiomer 1 with high enantioselectivity.…”