Directed evolution of a model primordial enzyme provides insights into the development of the genetic code.

The contemporary proteinogenic repertoire contains 20 amino acids with diverse functional groups and side chain geometries. Primordial proteins, in contrast, were presumably constructed from a subset of these building blocks. Subsequent expansion of the proteinogenic alphabet would have enhanced the...

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Main Authors: Manuel M Müller, Jane R Allison, Narupat Hongdilokkul, Laurent Gaillon, Peter Kast, Wilfred F van Gunsteren, Philippe Marlière, Donald Hilvert
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2013-01-01
Series:PLoS Genetics
Online Access:https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1003187&type=printable
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author Manuel M Müller
Jane R Allison
Narupat Hongdilokkul
Laurent Gaillon
Peter Kast
Wilfred F van Gunsteren
Philippe Marlière
Donald Hilvert
author_facet Manuel M Müller
Jane R Allison
Narupat Hongdilokkul
Laurent Gaillon
Peter Kast
Wilfred F van Gunsteren
Philippe Marlière
Donald Hilvert
author_sort Manuel M Müller
collection DOAJ
description The contemporary proteinogenic repertoire contains 20 amino acids with diverse functional groups and side chain geometries. Primordial proteins, in contrast, were presumably constructed from a subset of these building blocks. Subsequent expansion of the proteinogenic alphabet would have enhanced their capabilities, fostering the metabolic prowess and organismal fitness of early living systems. While the addition of amino acids bearing innovative functional groups directly enhances the chemical repertoire of proteomes, the inclusion of chemically redundant monomers is difficult to rationalize. Here, we studied how a simplified chorismate mutase evolves upon expanding its amino acid alphabet from nine to potentially 20 letters. Continuous evolution provided an enhanced enzyme variant that has only two point mutations, both of which extend the alphabet and jointly improve protein stability by >4 kcal/mol and catalytic activity tenfold. The same, seemingly innocuous substitutions (Ile→Thr, Leu→Val) occurred in several independent evolutionary trajectories. The increase in fitness they confer indicates that building blocks with very similar side chain structures are highly beneficial for fine-tuning protein structure and function.
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publishDate 2013-01-01
publisher Public Library of Science (PLoS)
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spelling doaj-art-0f687fdbf2764a9ca772733f50d682da2025-08-20T02:15:24ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042013-01-0191e100318710.1371/journal.pgen.1003187Directed evolution of a model primordial enzyme provides insights into the development of the genetic code.Manuel M MüllerJane R AllisonNarupat HongdilokkulLaurent GaillonPeter KastWilfred F van GunsterenPhilippe MarlièreDonald HilvertThe contemporary proteinogenic repertoire contains 20 amino acids with diverse functional groups and side chain geometries. Primordial proteins, in contrast, were presumably constructed from a subset of these building blocks. Subsequent expansion of the proteinogenic alphabet would have enhanced their capabilities, fostering the metabolic prowess and organismal fitness of early living systems. While the addition of amino acids bearing innovative functional groups directly enhances the chemical repertoire of proteomes, the inclusion of chemically redundant monomers is difficult to rationalize. Here, we studied how a simplified chorismate mutase evolves upon expanding its amino acid alphabet from nine to potentially 20 letters. Continuous evolution provided an enhanced enzyme variant that has only two point mutations, both of which extend the alphabet and jointly improve protein stability by >4 kcal/mol and catalytic activity tenfold. The same, seemingly innocuous substitutions (Ile→Thr, Leu→Val) occurred in several independent evolutionary trajectories. The increase in fitness they confer indicates that building blocks with very similar side chain structures are highly beneficial for fine-tuning protein structure and function.https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1003187&type=printable
spellingShingle Manuel M Müller
Jane R Allison
Narupat Hongdilokkul
Laurent Gaillon
Peter Kast
Wilfred F van Gunsteren
Philippe Marlière
Donald Hilvert
Directed evolution of a model primordial enzyme provides insights into the development of the genetic code.
PLoS Genetics
title Directed evolution of a model primordial enzyme provides insights into the development of the genetic code.
title_full Directed evolution of a model primordial enzyme provides insights into the development of the genetic code.
title_fullStr Directed evolution of a model primordial enzyme provides insights into the development of the genetic code.
title_full_unstemmed Directed evolution of a model primordial enzyme provides insights into the development of the genetic code.
title_short Directed evolution of a model primordial enzyme provides insights into the development of the genetic code.
title_sort directed evolution of a model primordial enzyme provides insights into the development of the genetic code
url https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1003187&type=printable
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