Srimathi, S and Jayaraman, G and Narayanan, P R (2006) Improved thermodynamic stability of subtilisin Carlsberg by covalent modification. Enzyme and Microbial Technology, 39 (2). pp. 301-307. ISSN Pritn: 0141-0229; Online: 1879-0909
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Abstract
The present work is aimed at improving the kinetic, thermal and thermodynamic stability of subtilisin Carlsberg (SCB) obtained from Bacillus licheniformis by means of simple, inexpensive but effective covalent coupling to oxidized sucrose polymers (OSP) of varying sizes (OSP400 and OSP70) as well as polyglutaraldehyde (PGA). In the presence of 10mM calcium the half-life of the enzyme at 60 ◦C increased by 6.06-fold, 5.20-fold and 2.92-fold when coupled with OSP400, OSP70 and PGA, respectively. Even in the absence of added calcium the stability against thermal inactivation was found to be greater for the modified enzymes as evident from the increase in the energy of activation for the inactivation process (Eai). Guanidium thiocyanate-induced unfolding indicated Cm values of 1.3 M, 1.8 M, 1.5Mand 1.4Mfor the native and enzymes modified with OSP400, OSP70 and PGA, respectively. Thermally induced unfolding was delayed for the modified enzymes as evident from the shift in Tm of 8.45 ◦C, 5.91 ◦C and 4.66 ◦C for OSP400, OSP70 and PGA modified enzymes. The results indicate that among the modifiers used OSP400 was most effective in stabilizing the enzyme and interestingly the increase in stability reported here is comparable to the most stabilized subtilisin variants obtained by site-directed mutagenesis.
| Item Type: | Article |
|---|---|
| Subjects: | Tuberculosis > Laboratory Research > Immunological |
| Divisions: | Basic Science Research > Immunology |
| Depositing User: | Dr. Rathinasabapati R |
| Date Deposited: | 19 Dec 2013 10:23 |
| Last Modified: | 09 Mar 2016 06:30 |
| URI: | http://eprints.nirt.res.in/id/eprint/781 |
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