IrrE Improves Organic Solvent Tolerance and Δ<sup>1</sup>-Dehydrogenation Productivity of <i>Arthrobacter simplex</i>

Song, Bo; Zhou, Qin; Xue, Haijie; Li, Jiajia; Zheng, Yue; Shen, Yanbing; Wang, Min; Luo, Jianmei

doi:10.1021/acs.jafc.8b01311

Cited by 21 publications

(33 citation statements)

References 38 publications

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“…A slight decrease in the biomass concentration, specific growth rate, glucose consumption rate, the final ethanol titer, ethanol productivity and ethanol yield were also observed in the strain BY4742/IrrE (Table 1). This result is different from previous studies in bacteria, where IrrE did not cause differences in the growth of strains under normal conditions [25,31]. However, the mutant of IrrE significantly increased strain biomass yield, glucose consumption rate, specific growth rate, final ethanol titer, ethanol productivity and ethanol yield (Fig.…”

Section: Evolved Irre To Improve Strain Tolerance To the Mixture Withcontrasting

confidence: 99%

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Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

Wang

et al. 2020

Biotechnol Biofuels

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Background Stress tolerance is one of the important desired microbial traits for industrial bioprocesses, and global regulatory protein engineering is an efficient approach to improve strain tolerance. In our study, IrrE, a global regulatory protein from the prokaryotic organism Deinococcus radiodurans, was engineered to confer yeast improved tolerance to the inhibitors in lignocellulose hydrolysates or high temperatures. Results Three IrrE mutations were developed through directed evolution, and the expression of these mutants could improve the yeast fermentation rate by threefold or more in the presence of multiple inhibitors. Subsequently, the tolerance to multiple inhibitors of single-site mutants based on the mutations from the variants were then evaluated, and 11 mutants, including L65P, I103T, E119V, L160F, P162S, M169V, V204A, R244G, Base 824 Deletion, V299A, and A300V were identified to be critical for the improved representative inhibitors, i.e., furfural, acetic acid and phenol (FAP) tolerance. Further studies indicated that IrrE caused genome-wide transcriptional perturbation in yeast, and the mutant I24 led to the rapid growth of Saccharomyces cerevisiae by primarily regulating the transcription level of transcription activators/factors, protecting the intracellular environment and enhancing the antioxidant capacity under inhibitor environments, which reflected IrrE plasticity. Meanwhile, we observed that the expression of the wild-type or mutant IrrE could also protect Saccharomyces cerevisiae from the damage caused by thermal stress. The recombinant yeast strains were able to grow with glucose at 42 ℃. Conclusions IrrE from Deinococcus radiodurans can be engineered as a tolerance-enhancer for Saccharomyces cerevisiae. Systematic research on the regulatory model and mechanism of a prokaryotic global regulatory factor IrrE to increase yeast tolerance provided valuable insights for the improvements in microbial tolerance to complex industrial stress conditions.

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Section: Evolved Irre To Improve Strain Tolerance To the Mixture Withcontrasting

confidence: 99%

“…The expression of the protein IrrE also protected Zymomonas mobilis against ethanol and acid stress [30]. The introduction of IrrE efficiently improved Arthrobacter simplex tolerance to organic solvents and various abiotic stresses as well [31]. Recently research exhibited that Sphingomonas sp.…”

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confidence: 99%

Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

Wang

et al. 2020

Biotechnol Biofuels

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“…Isolation of RNA was carried out according to the method described by our previous description [13]. The qRT-PCR analysis was performed according to the previously described method [37]. The primers for qRT-PCR were listed in Table 1.…”

Section: Quantitative Reverse Transcription-pcr (Qrt-pcr)mentioning

confidence: 99%

Improving phytosterol biotransformation at low nitrogen levels by enhancing the methylcitrate cycle with transcriptional regulators PrpR and GlnR of Mycobacterium neoaurum

et al. 2020

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Background: Androstenedione (AD) is an important steroid medicine intermediate that is obtained via the degradation of phytosterols by mycobacteria. The production process of AD is mainly the degradation of the phytosterol aliphatic side chain, which is accompanied by the production of propionyl CoA. Excessive accumulation of intracellular propionyl-CoA produces a toxic effect in mycobacteria, which restricts the improvement of production efficiency. The 2-methylcitrate cycle pathway (MCC) plays a significant role in the detoxification of propionyl-CoA in bacterial. The effect of the MCC on phytosterol biotransformation in mycobacteria has not been elucidated in detail. Meanwhile, reducing fermentation cost has always been an important issue to be solved in the optimizing of the bioprocess. Results: There is a complete MCC in Mycobacterium neoaurum (MNR), prpC, prpD and prpB in the prp operon encode methylcitrate synthase, methylcitrate dehydratase and methylisocitrate lyase involved in MCC, and PrpR is a specific transcriptional activator of prp operon. After the overexpression of prpDCB and prpR in MNR, the significantly improved transcription levels of prpC, prpD and prpB were observed. The highest conversion ratios of AD obtained by MNR-prpDBC and MNR-prpR increased from 72.3 ± 2.5% to 82.2 ± 2.2% and 90.6 ± 2.6%, respectively. Through enhanced the PrpR of MNR, the in intracellular propionyl-CoA levels decreased by 43 ± 3%, and the cell viability improved by 22 ± 1% compared to MNR at 96 h. The nitrogen transcription regulator GlnR repressed prp operon transcription in a nitrogen-limited medium. The glnR deletion enhanced the transcription level of prpDBC and the biotransformation ability of MNR. MNR-prpR/ΔglnR was constructed by the overexpression of prpR in the glnR-deleted strain showed adaptability to low nitrogen. The highest AD conversion ratio by MNR-prpR/ΔglnR was 92.8 ± 2.7% at low nitrogen level, which was 1.4 times higher than that of MNR.

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“…The expression of protein IrrE also protected Zymomonas mobilis against ethanol and acid stress [25]. The introduction of IrrE e ciently improved Arthrobacter simplex tolerance to organic solvents and various abiotic stresses as well [26]. Recently research exhibited that Sphingomonas sp.…”

Section: Introductionmentioning

confidence: 99%

Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

Wang

et al. 2020

Preprint

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Background: Stress tolerance is one of the important desired microbial traits for industrial bioprocess and global regulatory protein engineering is an efficient approach to improve strain tolerance. In our study, IrrE, a global regulatory protein from prokaryotic organism Deinococcus radiodurans, was engineered to confer yeast the improved tolerance to the inhibitors in lignocellulose hydrolysates or high temperature.Results: Three IrrE mutants were developed through directed evolution and the expression of these mutants could improve the yeast fermentation rate by 3- to 4-fold in the presence of multiple inhibitors. Subsequently, the tolerance to multiple inhibitors of single-site mutants based on the mutations from the variants was then evaluated, and eleven mutants including L65P, I103T, E119V, L160F, P162S, M169V, V204A, R244G, Base 824 Deletion, V299A and A300V were identified to be critical for the improved FAP tolerance. Further studies indicated that IrrE caused genome-wide transcriptional perturbation in yeast, and the mutant I24 led to the rapid growth of Saccharomyces cerevisiae mainly by regulating the transcription level of transcription activators/factors, protecting the intracellular environment and enhancing the antioxidant capacity under inhibitor environment, which reflected the plasticity of IrrE. Meanwhile, we observed that the expression of the wild-type or mutant IrrE could also protect Saccharomyces cerevisiae from the damage caused by thermal stress. The recombinant yeast strains were able to grow with glucose at 42 ºC.Conclusions: IrrE from Deinococcus radiodurans can be engineered as a tolerance-enhancer for Saccharomyces cerevisiae. Systematic research on the regulatory model and mechanism of a prokaryotic global regulatory factor IrrE to increase yeast tolerance provided valuable insights for the improvement of microbial tolerance to complex industrial stress conditions.

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IrrE Improves Organic Solvent Tolerance and Δ¹-Dehydrogenation Productivity of Arthrobacter simplex

Cited by 21 publications

References 38 publications

Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

Improving phytosterol biotransformation at low nitrogen levels by enhancing the methylcitrate cycle with transcriptional regulators PrpR and GlnR of Mycobacterium neoaurum

Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

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IrrE Improves Organic Solvent Tolerance and Δ1-Dehydrogenation Productivity of Arthrobacter simplex

Cited by 21 publications

References 38 publications

Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

Improving phytosterol biotransformation at low nitrogen levels by enhancing the methylcitrate cycle with transcriptional regulators PrpR and GlnR of Mycobacterium neoaurum

Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast

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IrrE Improves Organic Solvent Tolerance and Δ¹-Dehydrogenation Productivity of Arthrobacter simplex