Aqueous-like Activity of α-Chymotrypsin Dissolved in Nearly Anhydrous Organic Solvents

Journal of the American Chemical Society

Volumn 116, Issue 11, 1994, Pages 5009-5010

  Paradkar, Vikram M ^a   Dordick, Jonathan S ^a  

^a UNIVERSITY OF IOWA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

EID: 0027962947     PISSN: 00027863     EISSN: 15205126     Source Type: Journal    
DOI: 10.1021/ja00090a065     Document Type: Article

References (20)

1. See the following reviews and references therein: (a) Klibanov, A. M. Trends Biochem. Sci. 1989, 14, 141.
2. Dordick, J. S. Enzyme Microb. Technol. 1989, 11, 194.
3. Dordick, J. S. Biotechnol. Prog. 1992, 8, 259.
4. Zaks, A.; Russell, A. J. J. Biotechnol. 1988, 8, 259.
5. Inada, Y.; Katsunobu, T.; Yoshimoto, T.; Ajima, A.; Matsushima, A.; Saito, Y. Trends Biochem. Technol. 1986, 4, 190.
6. Arseguel, D.; Lattes, A.; Baboulene, M. Biocatalysis 1990, 3, 227.
7. Martinek, K.; Levashov, A. V.; Khmelnitsky, Yu. L.; Klyachko, N. L.; Berezin, I. V. Eur. J. Biochem. 1986, 155, 453.
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11. Paradkar, V. M.; Dordick, J. S. Biotechnol. Bioeng. 1994, 43, 529.
12. Zaks, A.; Klibanov, A. M. J. Am. Chem. Soc. 1986, 108, 2767.
13. Isooctane-soluble CT was titrated as follows: A 2-fold molar excess (to enzyme) of N-trans-cinnamoylimidazole (NTC) (Schonbaum, G. R.; Zerner, B.; Bender, M. L. J. Biol. Chem. 1961, 236, 2930) was dissolved in the solvent, and the enzyme solution was incubated at 25 °C for 90 min with mild shaking at 50 rpm. The CT was precipitated from the reaction mixture by addition of 1% (v/v) methanol, and the solids were centrifuged. The supernatant, containing unreacted NTC, was analyzed by gas chromatography.
14. The ionogenic state of an enzyme in organic media remains the same as in the last aqueous solution from which the enzyme is recovered (Zaks, A.; Klibanov, A. M. J. Biol. Chem. 1988, 263, 3194).
15. Khmelnitsky, Yu. L.; Welch, S. H.; Clark, D. S.; Dordick, J. S. J. Am. Chem. Soc. 1994, 116, 2647.
16. Gorman, L. S.; Dordick, J. S. Biotechnol. Bioeng. 1992, 39, 392.
17. Ryu, K.; Dordick, J.S. Biochemistry 1992, 31, 2588.
18. Wangikar, P. P.; Graycar, T. P.; Estell, D. A.; Clark, D. S.; Dordick, J. S. J. Am. Chem. Soc. 1993, 115, 12231.
19. Blow, D. M. Acc. Chem. Res. 1976, 9, 145.
20. Ala-Pro is an intermediate in the synthesis of the antihypertensive drug enalapril (Patchett, A. A.; et al. Nature 1980, 288, 280.)