6GVS

Engineered glycolyl-CoA reductase comprising 8 mutations with bound NADP+

Summary for 6GVS

• Entry DOI: 10.2210/pdb6gvs/pdb
• Descriptor: Aldehyde dehydrogenase, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, POTASSIUM ION, ... (4 entities in total)
• Functional Keywords: glycolyl-coa reductase, enzyme engineering, nadph, oxidoreductase
• Biological source: Rhodopseudomonas palustris BisB18; More
• Total number of polymer chains: 10
• Total formula weight: 560576.44

Authors

Zarzycki, J.,Trudeau, D.,Scheffen, M.,Erb, T.J.,Tawfik, D.S. (deposition date: 2018-06-21, release date: 2018-11-28, Last modification date: 2024-01-17)

Primary citation

Trudeau, D.L.,Edlich-Muth, C.,Zarzycki, J.,Scheffen, M.,Goldsmith, M.,Khersonsky, O.,Avizemer, Z.,Fleishman, S.J.,Cotton, C.A.R.,Erb, T.J.,Tawfik, D.S.,Bar-Even, A.
Design and in vitro realization of carbon-conserving photorespiration.
Proc. Natl. Acad. Sci. U.S.A., 115:E11455-E11464, 2018

PubMed Abstract: Photorespiration recycles ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) oxygenation product, 2-phosphoglycolate, back into the Calvin Cycle. Natural photorespiration, however, limits agricultural productivity by dissipating energy and releasing CO Several photorespiration bypasses have been previously suggested but were limited to existing enzymes and pathways that release CO Here, we harness the power of enzyme and metabolic engineering to establish synthetic routes that bypass photorespiration without CO release. By defining specific reaction rules, we systematically identified promising routes that assimilate 2-phosphoglycolate into the Calvin Cycle without carbon loss. We further developed a kinetic-stoichiometric model that indicates that the identified synthetic shunts could potentially enhance carbon fixation rate across the physiological range of irradiation and CO, even if most of their enzymes operate at a tenth of Rubisco's maximal carboxylation activity. Glycolate reduction to glycolaldehyde is essential for several of the synthetic shunts but is not known to occur naturally. We, therefore, used computational design and directed evolution to establish this activity in two sequential reactions. An acetyl-CoA synthetase was engineered for higher stability and glycolyl-CoA synthesis. A propionyl-CoA reductase was engineered for higher selectivity for glycolyl-CoA and for use of NADPH over NAD, thereby favoring reduction over oxidation. The engineered glycolate reduction module was then combined with downstream condensation and assimilation of glycolaldehyde to ribulose 1,5-bisphosphate, thus providing proof of principle for a carbon-conserving photorespiration pathway.

PubMed: 30459276
DOI: 10.1073/pnas.1812605115

Experimental method

X-RAY DIFFRACTION (2.579 Å)