Highly active rubiscos discovered by systematic interrogation of natural sequence diversity

Dan Davidi, Melina Shamshoum, Zhijun Guo, Yinon M. Bar‐On, Noam Prywes, Aia Oz, Jagoda Jablonska, Avi Flamholz, David G. Wernick, Niv Antonovsky, Lior Shachar, Dina Hochhauser, Yoav Peleg, Shira Albeck, Itai Sharon, Oliver Mueller‐Cajar, Ron Milo

Research output: Contribution to journalArticlepeer-review

Abstract

CO 2 is converted into biomass almost solely by the enzyme rubisco. The poor carboxylation properties of plant rubiscos have led to efforts that made it the most kinetically characterized enzyme, yet these studies focused on < 5% of its natural diversity. Here, we searched for fast‐carboxylating variants by systematically mining genomic and metagenomic data. Approximately 33,000 unique rubisco sequences were identified and clustered into ≈ 1,000 similarity groups. We then synthesized, purified, and biochemically tested the carboxylation rates of 143 representatives, spanning all clusters of form‐II and form‐II /III rubiscos. Most variants (> 100) were active in vitro , with the fastest having a turnover number of 22 ± 1 s−1—sixfold faster than the median plant rubisco and nearly twofold faster than the fastest measured rubisco to date. Unlike rubiscos from plants and cyanobacteria, the fastest variants discovered here are homodimers and exhibit a much simpler folding and activation kinetics. Our pipeline can be utilized to explore the kinetic space of other enzymes of interest, allowing us to get a better view of the biosynthetic potential of the biosphere.
Original languageEnglish
Article numbere104081
Number of pages11
JournalThe EMBO Journal
Volume39
Issue number18
Early online date5 Jun 2020
DOIs
StatePublished - 15 Sep 2020

All Science Journal Classification (ASJC) codes

  • General Immunology and Microbiology
  • General Biochemistry,Genetics and Molecular Biology
  • Molecular Biology
  • General Neuroscience

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