A map of the rubisco biochemical landscape
Abstract Rubisco is the primary CO2fixing enzyme of the biosphere yet has slow kinetics. The roles of evolution and chemical mechanism in constraining the sequence landscape of rubisco remain debated. In order to map sequence to function, we developed a massively parallel assay for rubisco using an engineeredE. coliwhere enzyme function is coupled to growth. By assaying >99% of single amino acid mutants across CO2concentrations, we inferred enzyme velocity and CO2affinity for thousands of substitutions. We identified many highly conserved positions that tolerate mutation and rare mutations that improve CO2affinity. These data suggest that non-trivial kinetic improvements are readily accessible and provide a comprehensive sequence-to-function mapping for enzyme engineering efforts..
| Medienart: |
|
|---|
Preprint| Erscheinungsjahr: |
2024 |
|---|---|
| Erschienen: |
2024 |
| Enthalten in: |
bioRxiv.org - (2024) vom: 15. Apr. Zur Gesamtaufnahme - year:2024 |
|---|
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Prywes, Noam [VerfasserIn] |
|---|
| Links: |
Volltext [kostenfrei] |
|---|
| Themen: |
|---|
| doi: |
10.1101/2023.09.27.559826 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
XBI040987043 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | XBI040987043 | ||
| 003 | DE-627 | ||
| 005 | 20240419090816.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231002s2024 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1101/2023.09.27.559826 |2 doi | |
| 035 | |a (DE-627)XBI040987043 | ||
| 035 | |a (biorXiv)10.1101/2023.09.27.559826 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Prywes, Noam |e verfasserin |0 (orcid)0000-0003-2526-2392 |4 aut | |
| 245 | 1 | 0 | |a A map of the rubisco biochemical landscape |
| 264 | 1 | |c 2024 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 520 | |a Abstract Rubisco is the primary CO2fixing enzyme of the biosphere yet has slow kinetics. The roles of evolution and chemical mechanism in constraining the sequence landscape of rubisco remain debated. In order to map sequence to function, we developed a massively parallel assay for rubisco using an engineeredE. coliwhere enzyme function is coupled to growth. By assaying >99% of single amino acid mutants across CO2concentrations, we inferred enzyme velocity and CO2affinity for thousands of substitutions. We identified many highly conserved positions that tolerate mutation and rare mutations that improve CO2affinity. These data suggest that non-trivial kinetic improvements are readily accessible and provide a comprehensive sequence-to-function mapping for enzyme engineering efforts. | ||
| 650 | 4 | |a Biology |7 (dpeaa)DE-84 | |
| 650 | 4 | |a 570 |7 (dpeaa)DE-84 | |
| 700 | 1 | |a Philips, Naiya R. |e verfasserin |0 (orcid)0000-0003-1836-5182 |4 aut | |
| 700 | 1 | |a Oltrogge, Luke M. |e verfasserin |0 (orcid)0000-0001-5716-9980 |4 aut | |
| 700 | 1 | |a Lindner, Sebastian |e verfasserin |4 aut | |
| 700 | 1 | |a Tsai, Yi-Chin Candace |e verfasserin |4 aut | |
| 700 | 1 | |a de Pins, Benoit |e verfasserin |0 (orcid)0000-0002-6779-8446 |4 aut | |
| 700 | 1 | |a Cowan, Aidan E. |e verfasserin |0 (orcid)0000-0003-2810-9318 |4 aut | |
| 700 | 1 | |a Taylor-Kearney, Leah J. |e verfasserin |0 (orcid)0000-0001-7530-7670 |4 aut | |
| 700 | 1 | |a Chang, Hana A. |e verfasserin |4 aut | |
| 700 | 1 | |a Hall, Laina N. |e verfasserin |0 (orcid)0000-0003-3135-332X |4 aut | |
| 700 | 1 | |a Bellieny-Rabelo, Daniel |e verfasserin |4 aut | |
| 700 | 1 | |a Nisonoff, Hunter M. |e verfasserin |4 aut | |
| 700 | 1 | |a Weissman, Rachel F. |e verfasserin |4 aut | |
| 700 | 1 | |a Flamholz, Avi I. |e verfasserin |4 aut | |
| 700 | 1 | |a Ding, David |e verfasserin |0 (orcid)0000-0001-6105-2259 |4 aut | |
| 700 | 1 | |a Bhatt, Abhishek Y. |e verfasserin |4 aut | |
| 700 | 1 | |a Shih, Patrick M. |e verfasserin |0 (orcid)0000-0002-2119-3345 |4 aut | |
| 700 | 1 | |a Mueller-Cajar, Oliver |e verfasserin |4 aut | |
| 700 | 1 | |a Milo, Ron |e verfasserin |0 (orcid)0000-0003-1641-2299 |4 aut | |
| 700 | 1 | |a Savage, David F. |e verfasserin |0 (orcid)0000-0003-0042-2257 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t bioRxiv.org |g (2024) vom: 15. Apr. |
| 773 | 1 | 8 | |g year:2024 |g day:15 |g month:04 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1101/2023.09.27.559826 |x 0 |z kostenfrei |3 Volltext |
| 912 | |a GBV_XBI | ||
| 951 | |a AR | ||
| 952 | |j 2024 |b 15 |c 04 | ||