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
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
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 PDB Entries

Structure ID Release Date Resolution Structure Title
1GOI 2001-11-15 1.45 Crystal structure of the D140N mutant of chitinase B from Serratia marcescens at 1.45 A resolution
3WD0 2013-09-18 1.7 Serratia marcescens Chitinase B, tetragonal form
1O6I 2003-03-30 1.7 Chitinase B from Serratia marcescens complexed with the catalytic intermediate mimic cyclic dipeptide CI4.
1OGB 2004-04-27 1.85 Chitinase b from Serratia marcescens mutant D142N
1OGG 2004-04-27 1.97 chitinase b from serratia marcescens mutant d142n in complex with inhibitor allosamidin
3WD4 2013-09-18 2.0 Serratia marcescens Chitinase B complexed with azide inhibitor and quinoline compound
1H0I 2002-06-27 2.0 Complex of a chitinase with the natural product cyclopentapeptide argifin from Gliocladium
4Z2I 2015-07-01 2.0 Serratia marcescens Chitinase B complexed with macrolide inhibitor 30
1H0G 2002-06-27 2.0 Complex of a chitinase with the natural product cyclopentapeptide argadin from Clonostachys
3WD3 2013-09-18 2.2 Serratia marcescens Chitinase B complexed with azide inhibitor
3WD2 2013-09-18 2.2 Serratia marcescens Chitinase B complexed with azide inhibitor
1E6N 2001-06-22 2.25 Chitinase B from Serratia marcescens inactive mutant E144Q in complex with N-acetylglucosamine-pentamer
3WD1 2013-09-18 2.3 Serratia marcescens Chitinase B complexed with syn-triazole inhibitor
4Z2K 2015-07-01 2.3 Serratia marcescens Chitinase B complexed with macrolide inhibitor 32
4Z2L 2015-07-01 2.3 Serratia marcescens Chitinase B complexed with macrolide inhibitor 33
4Z2H 2015-07-01 2.35 Serratia marcescens Chitinase B complexed with macrolide inhibitor 29
4Z2J 2015-07-01 2.6 Serratia marcescens Chitinase B complexed with macrolide inhibitor 31
4Z2G 2015-07-01 2.6 Serratia marcescens Chitinase B complexed with macrolide inhibitor 26

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
98.4 Chitinase P11797 CHIB_SERMA