Semirational Design Based on Consensus Sequences to Balance the Enzyme Activity-Stability Trade-Off
In this study, the phenomenon of the stability-activity trade-off, which is increasingly recognized in enzyme engineering, was explored. Typically, enhanced stability in enzymes correlates with diminished activity. Utilizing Rosa roxburghii copper-zinc superoxide dismutase (RrCuZnSOD) as a model, single-site mutations were introduced based on a semirational design derived from consensus sequences. The initial set of mutants was selected based on activity, followed by combinatorial mutation. This approach yielded two double-site mutants, D25/A115T (18,688 ± 206 U/mg) and A115T/S135P (18,095 ± 1556 U/mg), exhibiting superior enzymatic properties due to additive and synergistic effects. These mutants demonstrated increased half-lives (T1/2) at 80 °C by 1.2- and 1.6-fold, respectively, and their melting temperatures (Tm) rose by 3.4 and 2.5 °C, respectively, without any loss in activity relative to the wild type. Via an integration of structural analysis and molecular dynamics simulations, we elucidated the underlying mechanism facilitating the concurrent enhancement of both thermostability and enzymatic activity.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2024 |
|---|---|
| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:72 |
|---|---|
| Enthalten in: |
Journal of agricultural and food chemistry - 72(2024), 12 vom: 27. März, Seite 6454-6462 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Zhao, Yang [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Consensus sequences |
|---|
| Anmerkungen: |
Date Completed 28.03.2024 Date Revised 28.03.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1021/acs.jafc.3c08620 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM369676777 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM369676777 | ||
| 003 | DE-627 | ||
| 005 | 20240329000757.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 240313s2024 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1021/acs.jafc.3c08620 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1353.xml |
| 035 | |a (DE-627)NLM369676777 | ||
| 035 | |a (NLM)38477968 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Zhao, Yang |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Semirational Design Based on Consensus Sequences to Balance the Enzyme Activity-Stability Trade-Off |
| 264 | 1 | |c 2024 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 28.03.2024 | ||
| 500 | |a Date Revised 28.03.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a In this study, the phenomenon of the stability-activity trade-off, which is increasingly recognized in enzyme engineering, was explored. Typically, enhanced stability in enzymes correlates with diminished activity. Utilizing Rosa roxburghii copper-zinc superoxide dismutase (RrCuZnSOD) as a model, single-site mutations were introduced based on a semirational design derived from consensus sequences. The initial set of mutants was selected based on activity, followed by combinatorial mutation. This approach yielded two double-site mutants, D25/A115T (18,688 ± 206 U/mg) and A115T/S135P (18,095 ± 1556 U/mg), exhibiting superior enzymatic properties due to additive and synergistic effects. These mutants demonstrated increased half-lives (T1/2) at 80 °C by 1.2- and 1.6-fold, respectively, and their melting temperatures (Tm) rose by 3.4 and 2.5 °C, respectively, without any loss in activity relative to the wild type. Via an integration of structural analysis and molecular dynamics simulations, we elucidated the underlying mechanism facilitating the concurrent enhancement of both thermostability and enzymatic activity | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a consensus sequences | |
| 650 | 4 | |a molecular dynamics simulation | |
| 650 | 4 | |a protein engineering | |
| 650 | 4 | |a stability-activity trade-off | |
| 650 | 4 | |a thermostability | |
| 700 | 1 | |a Chen, Kun |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, Haixia |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Yongtao |e verfasserin |4 aut | |
| 700 | 1 | |a Liao, Xiaojun |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Journal of agricultural and food chemistry |d 1967 |g 72(2024), 12 vom: 27. März, Seite 6454-6462 |w (DE-627)NLM000005665 |x 1520-5118 |7 nnns |
| 773 | 1 | 8 | |g volume:72 |g year:2024 |g number:12 |g day:27 |g month:03 |g pages:6454-6462 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1021/acs.jafc.3c08620 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 72 |j 2024 |e 12 |b 27 |c 03 |h 6454-6462 | ||