Stabilization of a chitinase from Serratia marcescens by Gly-->Ala and Xxx-->Pro mutations.


Abstract

This paper describes attempts to increase the kinetic stability of chitinase B from Serratia marcescens (ChiB) by the introduction of semi-automatically designed rigidifying mutations of the Gly-->Ala and Xxx-->Pro type. Of 15 single mutants, several displayed significant increases in thermal stability, whereas most mutants showed minor effects. All mutations with non-marginal effects on stability clustered in a limited, surface-exposed region of the enzyme, indicating that this region is involved in a partial unfolding process that triggers irreversible thermal inactivation (aggregation). A double mutant containing two stabilizing mutations in this region (G188A, A234P) displayed a 10-fold increase in half-life at 57 degrees C and a 4.2 degrees C increase in apparent T(m). These results show that entropic stabilization works well for ChiB and they pinpoint a region whose unfolding may be crucial for the kinetic stability of this enzyme. Study holds ProTherm entries: 17397, 17398 Extra Details: Chitinase; proline; thermal stability; unfolding

Submission Details

ID: 6WWbq7g63

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:49 p.m.

Version: 1

Publication Details
GĂ„seidnes S;Synstad B;Jia X;Kjellesvik H;Vriend G;Eijsink VG,Protein Eng. (2003) Stabilization of a chitinase from Serratia marcescens by Gly-->Ala and Xxx-->Pro mutations. PMID:14631073
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