Induced Stereoselectivity and Substrate Selectivity of Bio-Imprinted α-Chymotrypsin in Anhydrous Organic Media

Journal of the American Chemical Society

Volumn 113, Issue 24, 1991, Pages 9366-9368

  Ståhl, Marianne ^a   Jeppsson Wistrand, U ^a   Mansson, Mats Olie ^a   Klaus, Mosbach ^a  

^a LUND UNIVERSITY   (Sweden)

Author keywords

[No Author keywords available]

Indexed keywords

EID: 0000983403     PISSN: 00027863     EISSN: 15205126     Source Type: Journal    
DOI: 10.1021/ja00024a051     Document Type: Article

References (15)

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3. Ståhl, M.; Månsson, M. O.; Mosbach, K. Swedish patent application no. 89040215-4 1989.
4. Braco, L.; Dabulis, K.; Klibanov, A. M. Proc. Natl. Acad. Sci. U.S.A. 1990,87, 274–277.
5. Ståhl, M.; Månsson, M. O.; Mosbach, K. Biotechnol. Lett. 1990, 12, 161–166.
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7. Ekberg, B.; Mosbach, K. TIBTECH 1989, 7, 92–96.
8. Initial water content of the reaction mixture was 10 mM and after equilibration with the enzyme 20 mM, indicating that water had been removed from the enzyme and into the solvent (measured using the Fisher method). Water content of the dried enzyme (measured gravimetrically) was 12.2% (w/w).
9. α-Chymotrypsin (30 mg) was dissolved in 1 mL of 10 mM sodium phosphate buffer pH 7.8 containing the amino acid derivative (20 mM). The enzyme solution was cooled to 0 °C and equilibrated for 30 min, and then 4 mL of 1-propanol (−20 °C) was added. The precipitate formed was recovered by centrifugation after 30 min on ice. After three washes with 1 -propanol the precipitate was dried under vacuum. The dried precipitate contained 3.5 mol amino acid derivative/mol α-chymotrypsin.
10. Reslow, M.; Adlercreutz, P.; Mattiasson, B. Appl. Microbiol. Biotechnol. 1987, 26, 1–8.
11. Zaks, A.; Klibanov, A. M. J. Biol. Chem. 1988, 263, 8017–8021.
12. Kitaguchi, H.; Klibanov, A. M. J. Am. Chem. Soc. 1989, 111, 9272–9273.
13. Bio-imprinted (8 mg) α-chymotrypsin was suspended in 2 mL of cy-clohexane and sonicated (1 min). Ethanol (0.5 ml) with 0.1 M N-acetylated amino acid was added. Product formation was followed with HPLC. Optical purity of the product was determined to be at least 98% as judged by measuring the specific rotation polarimetricalty.
14. Schonbaum, G. R.; Zerner, B.; Bender, M. L. J. Biol. Chem. 1961, 236, 2930–2935.
15. Gold, A. M. Methods Enzymol. 1967, 11, 706–711.