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
Gåseidnes 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:14631073Additional Information
Study Summary
Structure view and single mutant data analysis
Study data
| Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
|---|
Data Distribution
Studies with similar sequences (approximate matches)
Correlation with other assays (exact sequence matches)
Relevant UniProtKB Entries
| Percent Identity | Matching Chains | Protein | Accession | Entry Name |
|---|---|---|---|---|
| 98.4 | Chitinase | P11797 | CHIB_SERMA |