Assessing the potential of mutational strategies to elicit new phenotypes in industrial strains
- 19 February 2008
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 105 (7), 2319-2324
- https://doi.org/10.1073/pnas.0712177105
Abstract
Industrial strains have been traditionally improved by rational approaches and combinatorial methods involving mutagenesis and selection. Recently, other methods have emerged, such as the use of artificial transcription factors and engineering of the native ones. As methods for generating genetic diversity continue to proliferate, the need for quantifying phenotypic diversity and, hence, assessing the potential of various genetic libraries for strain improvement becomes more pronounced. Here, we present a metric based on the quantification of phenotypic diversity, using Lactobacillus plantarum as a model organism. We found that phenotypic diversity can be introduced by mutagenesis of the principal sigma factor, that this diversity can be modulated by tuning the sequence diversity, and that this method compares favorably with commonly used protocols for chemical mutagenesis. The results of the diversity metric here developed also correlated well with the probability of finding improved mutants in the different libraries, as determined by recursive screening under stress. In addition, we subjected our libraries to lactic and inorganic acids and found strains with improved growth in both conditions, with a concomitant increase in lactate productivity.This publication has 44 references indexed in Scilit:
- Identification and Use of Zinc Finger Transcription Factors That Increase Production of Recombinant Proteins in Yeast and Mammalian CellsBiotechnology Progress, 2008
- Divergence of Transcription Factor Binding Sites Across Related Yeast SpeciesScience, 2007
- Novel Redox Potential-Based Screening Strategy for Rapid Isolation of Klebsiella pneumoniae Mutants with Enhanced 1,3-Propanediol-Producing CapabilityApplied and Environmental Microbiology, 2007
- The cyanobacterial principal σ factor region 1.1 is involved in DNA-binding in the free form and in transcription activity as holoenzymeFEBS Letters, 2006
- Why High-error-rate Random Mutagenesis Libraries are Enriched in Functional and Improved ProteinsJournal of Molecular Biology, 2005
- Stochasticity in gene expression: from theories to phenotypesNature Reviews Genetics, 2005
- Phenotypic alteration of eukaryotic cells using randomized libraries of artificial transcription factorsNature Biotechnology, 2003
- Possible intermediate steps in the evolution of a prokaryotic developmental systemProceedings. Biological sciences, 1991
- Altered promoter recognition by mutant forms of the σ70 subunit of Escherichia coli RNA polymeraseJournal of Molecular Biology, 1989
- A mutant Escherichia coliσ70 subunit of RNA polymerase with altered promoter specificityJournal of Molecular Biology, 1989