dc.contributor.authorChong, Huiqing
dc.contributor.authorGeng, Hefang
dc.contributor.authorZhang, Hongfang
dc.contributor.authorSong, Hao
dc.contributor.authorHuang, Lei
dc.contributor.authorJiang, Rongrong
dc.date.accessioned2014-04-07T05:04:16Z
dc.date.available2014-04-07T05:04:16Z
dc.date.copyright2013en_US
dc.date.issued2013
dc.identifier.citationChong, H., Geng, H., Zhang, H., Song, H., Huang, L., & Jiang, R. (2014). Enhancing E. coli isobutanol tolerance through engineering its global transcription factor cAMP receptor protein (CRP). Biotechnology and Bioengineering, 111(4), 700-708.en_US
dc.identifier.issn0006-3592en_US
dc.identifier.urihttp://hdl.handle.net/10220/19150
dc.description.abstractThe limited isobutanol tolerance of Escherichia coli is a major drawback during fermentative isobutanol production. Different from classical strain engineering approaches, this work was initiated to improve E. coli isobutanol tolerance from its transcriptional level by engineering its global transcription factor cAMP receptor protein (CRP). Random mutagenesis libraries were generated by error-prone PCR of crp, and the libraries were subjected to isobutanol stress for selection. Variant IB2 (S179P, H199R) was isolated and exhibited much better growth (0.18 h−1) than the control (0.05 h−1) in 1.2% (v/v) isobutanol (9.6 g/L). Genome-wide DNA microarray analysis revealed that 58 and 308 genes in IB2 had differential expression (>2-fold, p < 0.05) in the absence and presence of 1% (v/v) isobutanol, respectively. When challenged with isobutanol, genes related to acid resistance (gadABCE, hdeABD), nitrate reduction (narUZYWV), flagella and fimbrial activity (lfhA, yehB, ycgR, fimCDF), and sulfate reduction and transportation (cysIJH, cysC, cysN) were the major functional groups that were up-regulated, whereas most of the down-regulated genes were enzyme (tnaA) and transporters (proVWX, manXYZ). As demonstrated by single-gene knockout experiments, gadX, nirB, rhaS, hdeB, and ybaS were found associated with strain isobutanol resistance. The intracellular reactive oxygen species (ROS) level in IB2 was only half of that of the control when facing stress, indicating that IB2 can withstand toxic isobutanol much better than the control. Biotechnol. Biotechnol.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesBiotechnology and bioengineeringen_US
dc.rights© 2013 Wiley Periodicals, Inc.en_US
dc.subjectDRNTU::Engineering::Bioengineering
dc.titleEnhancing E. coli isobutanol tolerance through engineering its global transcription factor cAMP receptor protein (CRP)en_US
dc.typeJournal Article
dc.contributor.schoolSchool of Chemical and Biomedical Engineeringen_US
dc.identifier.doihttp://dx.doi.org/10.1002/bit.25134


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