There's No Industrial Biocatalyst Like Hydrolase:  Development of Scalable Enantioselective Processes Using Hydrolytic Enzymes1

• Published in: Organic process research & development Vol. 7; no. 3; pp. 289 - 296
• Main Authors: Chikusa, Yasuo, Hirayama, Yoshihiro, Ikunaka, Masaya, Inoue, Toru, Kamiyama, Shunji, Moriwaki, Masafumi, Nishimoto, Yukifumi, Nomoto, Fumiki, Ogawa, Kazuo, Ohno, Tomoyasu, Otsuka, Koutaro, Sakota, Akiko K, Shirasaka, Naoki, Uzura, Atsuko, Uzura, Kensaku
• Format: Journal Article
• Language: English
• Published: American Chemical Society 16.05.2003
• ISSN: 1083-6160; 1520-586X
• DOI: 10.1021/op034014b

Chiral, racemic esters, ethyl (±)-tetrahydrofuran-2-carboxylate 4c, methyl (±)-α-phenylpropionate 9b, methyl (±)-5,5-dimethyl-1,3-thiazolidine-4-carboxylate 12a, 2-methoxyethyl (±)-1-(4-tert-butylphenyl)-2-oxopyrrolidine-4-carboxylate 15a, (±)-1-benzyloxy-3-chloropropan-2-yl hydrogen succinate 18c, and (±)-3-butyryloxyquinuclidinium butyrate [(±)-20b·n-PrCO2H], are resolved kinetically by enantioselective hydrolysis catalyzed by an Aspergillus melleus protease [E = 60; 93.9% ee and 35% yield for (R)-tetrahydrofuran-2-carboxylic acid 4a], a Klebsiella oxytoca hydrolase [E > 200; 99.5% ee and 36% yield for (S)-α-phenylpropionic acid 9a], a K. oxytoca hydrolase [E = 145; 97.7% ee and 34% yield for (R)-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid 12b], a Bacillus brevis protease [E = 77; 99% ee and 45% yield for (S)-15a], a Serratia marcescence esterase [E = 49; 99% ee and 43% yield for (S)-18c], and an A. melleus protease [E = 96; 96% ee and 42% yield for (R)-20b], respectively. Each enzymatic process is discussed with focus on the following tactical issues:  (1) identification of a hydrolase with high enantioselectivity, (2) substrate concentrations not less than 1 M that allow for industrially viable volume efficiency (space−time yield), (3) product separation by partition between organic and aqueous phases, and (4) alleviation of a hydrolysate inhibiting the enzymatic activity.