| Chapter title |
A Computational Library Design Protocol for Rapid Improvement of Protein Stability: FRESCO
|
|---|---|
| Chapter number | 5 |
| Book title |
Protein Engineering
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7366-8_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7364-4, 978-1-4939-7366-8
|
| Authors |
Hein J. Wijma, Maximilian J. L. J. Fürst, Dick B. Janssen, Wijma, Hein J., Fürst, Maximilian J. L. J., Janssen, Dick B. |
| Abstract |
The ability to stabilize enzymes and other proteins has wide-ranging applications. Most protocols for enhancing enzyme stability require multiple rounds of high-throughput screening of mutant libraries and provide only modest improvements of stability. Here, we describe a computational library design protocol that can increase enzyme stability by 20-35 °C with little experimental screening, typically fewer than 200 variants. This protocol, termed FRESCO, scans the entire protein structure to identify stabilizing disulfide bonds and point mutations, explores their effect by molecular dynamics simulations, and provides mutant libraries with variants that have a good chance (>10%) to exhibit enhanced stability. After experimental verification, the most effective mutations are combined to produce highly robust enzymes. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 95 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 30 | 32% |
| Researcher | 13 | 14% |
| Student > Bachelor | 9 | 9% |
| Student > Master | 3 | 3% |
| Other | 2 | 2% |
| Other | 6 | 6% |
| Unknown | 32 | 34% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 32 | 34% |
| Agricultural and Biological Sciences | 10 | 11% |
| Chemistry | 9 | 9% |
| Chemical Engineering | 3 | 3% |
| Engineering | 2 | 2% |
| Other | 6 | 6% |
| Unknown | 33 | 35% |