Genome shuffling of Lactobacillus for improved acid tolerance

Fermentation-based bioprocesses rely extensively on strain improvement for commercialization. Whole-cell biocatalysts are commonly limited by low tolerance of extreme process conditions such as temperature, pH, and solute concentration. Rational approaches to improving such complex phenotypes lack g...

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Bibliographic Details
Published in:Nature biotechnology 2002-07, Vol.20 (7), p.707-712
Main Authors: del Cardayré, Stephen, Gavrilovic, Vesna, Ryan, Chris M, Patnaik, Ranjan, Louie, Susan, Stemmer, Willem P.C, Perry, Kim
Format: Article
Language:English
Subjects:
Agriculture
Biocatalysts
Bioinformatics
Biological and medical sciences
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
Catalysis
Cell fusion (prokaryotes)
Directed Molecular Evolution - methods
Evolution
Evolution, Molecular
Fermentation
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Bacterial
Genetic engineering
Genetic Enhancement - methods
Genetic technics
Genetic Variation
Genome, Bacterial
Genomes
Lactic Acid - biosynthesis
Lactobacillus - enzymology
Lactobacillus - genetics
Lactobacillus - growth & development
Life Sciences
Methods. Procedures. Technologies
Microorganisms
Molecular Biology - methods
Mutation - genetics
Organisms, Genetically Modified - genetics
Organisms, Genetically Modified - metabolism
Protons
Recombination, Genetic
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Summary:Fermentation-based bioprocesses rely extensively on strain improvement for commercialization. Whole-cell biocatalysts are commonly limited by low tolerance of extreme process conditions such as temperature, pH, and solute concentration. Rational approaches to improving such complex phenotypes lack good models and are especially difficult to implement without genetic tools. Here we describe the use of genome shuffling to improve the acid tolerance of a poorly characterized industrial strain of Lactobacillus . We used classical strain-improvement methods to generate populations with subtle improvements in pH tolerance, and then shuffled these populations by recursive pool-wise protoplast fusion. We identified new shuffled lactobacilli that grow at substantially lower pH than does the wild-type strain on both liquid and solid media. In addition, we identified shuffled strains that produced threefold more lactic acid than the wild type at pH 4.0. Genome shuffling seems broadly useful for the rapid evolution of tolerance and other complex phenotypes in industrial microorganisms.
ISSN:1087-0156
1546-1696
DOI:10.1038/nbt0702-707