ArticlePloS one2014
Mutagenic Organized Recombination Process by Homologous IN vivo Grouping (MORPHING) for directed enzyme evolution.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 62 citations in OpenAlex.
- Identification of improved signal peptides for heterologous expression in Saccharomyces using a screen that exploits Gaussia luciferase.Scientific reports · 2025Article
- Enzyme Engineering: Performance Optimization, Novel Sources, and Applications in the Food Industry.Foods (Basel, Switzerland) · 2024Review
- A primer to directed evolution: current methodologies and future directions.RSC chemical biology · 2023Review
- Recombination of Single Beneficial Substitutions Obtained from Protein Engineering by Computer-Assisted Recombination (CompassR).Methods in molecular biology (Clifton, N.J.) · 2022Article
- A modular two yeast species secretion system for the production and preparative application of unspecific peroxygenases.Communications biology · 2021Article
- Simultaneous directed evolution of coupled enzymes for efficient asymmetric synthesis ofApplied and environmental microbiology · 2021Article
- Identification and Expression of New Unspecific Peroxygenases - Recent Advances, Challenges and Opportunities.Frontiers in bioengineering and biotechnology · 2021Review
- Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases.Applied and environmental microbiology · 2020Article
- Computer-Assisted Recombination (CompassR) Teaches us How to Recombine Beneficial Substitutions from Directed Evolution Campaigns.Chemistry (Weinheim an der Bergstrasse, Germany) · 2020Article
- Directed evolution of the aryl-alcohol oxidase: Beyond the lab bench.Computational and structural biotechnology journal · 2020Review
- Evolving a Peptide: Library Platforms and Diversification Strategies.International journal of molecular sciences · 2019Review
- Directed -in vitro- evolution of Precambrian and extant Rubiscos.Scientific reports · 2018Article
- Modern methods for laboratory diversification of biomolecules.Current opinion in chemical biology · 2017Review
- Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening.Journal of visualized experiments : JoVE · 2016Article
- Focused Directed Evolution of Aryl-Alcohol Oxidase in Saccharomyces cerevisiae by Using Chimeric Signal Peptides.Applied and environmental microbiology · 2015Article
- Directed evolution of unspecific peroxygenase from Agrocybe aegerita.Applied and environmental microbiology · 2014Article
- Article
Corrections and comments
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Authors and funding
4 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Approaches that depend on directed evolution require reliable methods to generate DNA diversity so that mutant libraries can focus on specific target regions. We took advantage of the high frequency of homologous DNA recombination in Saccharomyces cerevisiae to develop a strategy for domain mutagenesis aimed at introducing and in vivo recombining random mutations in defined segments of DNA. Mutagenic Organized Recombination Process by Homologous IN vivo Grouping (MORPHING) is a one-pot random mutagenic method for short protein regions that harnesses the in vivo recombination apparatus of yeast. Using this approach, libraries can be prepared with different mutational loads in DNA segments of less than 30 amino acids so that they can be assembled into the remaining unaltered DNA regions in vivo with high fidelity. As a proof of concept, we present two eukaryotic-ligninolytic enzyme case studies: i) the enhancement of the oxidative stability of a H2O2-sensitive versatile peroxidase by independent evolution of three distinct protein segments (Leu28-Gly57, Leu149-Ala174 and Ile199-Leu268); and ii) the heterologous functional expression of an unspecific peroxygenase by exclusive evolution of its native 43-residue signal sequence.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.