15 research outputs found
Directed -in vitro- evolution of Precambrian and extant Rubiscos
Rubisco is an ancient, catalytically conserved yet slow enzyme, which plays a central role in the
biosphere’s carbon cycle. The design of Rubiscos to increase agricultural productivity has hitherto
relied on the use of in vivo selection systems, precluding the exploration of biochemical traits that are
not wired to cell survival. We present a directed -in vitro- evolution platform that extracts the enzyme
from its biological context to provide a new avenue for Rubisco engineering. Precambrian and extant
form II Rubiscos were subjected to an ensemble of directed evolution strategies aimed at improving
thermostability. The most recent ancestor of proteobacteria -dating back 2.4 billion years- was uniquely
tolerant to mutagenic loading. Adaptive evolution, focused evolution and genetic drift revealed a
panel of thermostable mutants, some deviating from the characteristic trade-offs in CO2-fixing speed
and specificity. Our findings provide a novel approach for identifying Rubisco variants with improved
catalytic evolution potential.This work was supported by the REPSOL Research contracts Rubolution (RC020401120018), Rubolution 2.0 (RC
020401140042), the CSIC project PIE-201780E043 and the Australian Research Council grant CE140100015