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

Exact results for N = 2 supersymmetric gauge theories on compact toric manifolds and equivariant Donaldson invariants

We provide a contour integral formula for the exact partition function of N = 2 supersymmetric U(N) gauge theories on compact toric four-manifolds by means of supersymmetric localisation. We perform the explicit evaluation of the contour integral for U(2) N = 2 17 theory on CP2 for all instanton numbers. In the zero mass case, corresponding to the N = 4 supersymmetric gauge theory, we obtain the generating function of the Euler characteristics of instanton moduli spaces in terms of mock-modular forms. In the decoupling limit of infinite mass we find that the generating function of local and surface observables computes equivariant Donaldson invariants, thus proving in this case a longstanding conjecture by N. Nekrasov. In the case of vanishing first Chern class the resulting equivariant Donaldson polynomials are new. \ua9 2016, The Author(s)