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Identification and reconstitution of genetic regulatory networks for improved microbial tolerance to isooctane.

Molecular bioSystems (2012-02-14)
Aram Kang, Matthew Wook Chang
ABSTRACT

Microbial tolerance to hydrocarbons has been studied in an effort to improve the productivity of biochemical processes and to enhance the efficiency of hydrocarbon bioremediation. Despite these studies, few attempts have been made to design rational strategies to improve microbial tolerance to hydrocarbons. Herein, we present an engineering framework that enables us to harness our understanding of genetic regulatory networks to improve hydrocarbon tolerance. In this study, isooctane was used as a representative hydrocarbon due to its use in petroleum refining and in biochemical processes. To increase isooctane tolerance, we first identified essential transcriptional determinants and genetic regulatory networks underlying cellular responses to isooctane in Escherichia coli using genome-wide microarray analysis. Based on functional transcriptome and bioinformatics analysis, a range of combinations of transcription factors whose activity was predictably perturbed by isooctane were knocked out and overexpressed to reconstitute the regulatory networks. We demonstrated that the reconstitution of the regulatory networks led to a significant improvement in isooctane tolerance, and especially, engineered E. coli strains lacking and overexpressing some of the perturbed transcription factors showed 3- to 5-fold improvement. This microbe with high tolerance to isooctane can be harnessed for biochemical processes, fuel oil bioremediation and metabolic engineering for biofuel production. Furthermore, we envision that the engineering framework employed to improve the tolerance in this study can be exploited for developing other microbes with desired phenotypes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2,2,4-三甲基戊烷, anhydrous, 99.8%
Sigma-Aldrich
2,2,4-三甲基戊烷, ACS reagent, ≥99.0%
Sigma-Aldrich
2,2,4-三甲基戊烷, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.5%
Sigma-Aldrich
2,2,4-三甲基戊烷, suitable for HPLC, ≥99%
Sigma-Aldrich
2,2,4-三甲基戊烷, puriss. p.a., ≥99.5% (GC)
Supelco
2,2,4-三甲基戊烷, analytical standard
Sigma-Aldrich
2,2,4-三甲基戊烷, ReagentPlus®, ≥99%
Sigma-Aldrich
2,2,4-三甲基戊烷, puriss. p.a., ACS reagent, ≥99.5% (GC)
Supelco
密度标准品 692kg/m3, H&D Fitzgerald Ltd. Quality