Rational Synthetic Combination Genetic Devices Boosting High Temperature Ethanol Fermentation

Overview

Journal Synth Syst Biotechnol

Specialty Biotechnology

Date 2017 Oct 25

PMID 29062969

Citations 4

Authors

Huan Sun

Haiyang Jia

Jun Li

Xudong Feng

Yueqin Liu

Xiaohong Zhou

Chun Li

Affiliations

Soon will be listed here.

Abstract

The growth and production of yeast in the industrial fermentation are seriously restrained by heat stress and exacerbated by heat induced oxidative stress. In this study, a novel synthetic biology approach was developed to globally boost the viability and production ability of at high temperature through rationally designing and combing heat shock protein (HSP) and superoxide dismutase (SOD) genetic devices to ultimately synergistically alleviate both heat stress and oxidative stress. HSP and SOD from extremophiles were constructed to be different genetic devices and they were preliminary screened by heat resistant experiments and anti-oxidative experiments, respectively. Then in order to customize and further improve thermotolerance of , the HSP genetic device and SOD genetic device were rationally combined. The results show the simply assemble of the same function genetic devices to solve heat stress or oxidative stress could not enhance the thermotolerance considerably. Only with the combination genetic device (FBA1p--FBA1p-) solving both stress showed 250% better thermotolerance than the control and displayed further 55% enhanced cell density compared with the strains with single FBA1p- or FBA1p- at 42 °C. Then the most excellent combination genetic device was introduced into lab and industrial for ethanol fermentation. The ethanol yields of the two strains were increased by 20.6% and 26.3% compared with the control under high temperature, respectively. These results indicate synergistically defensing both heat stress and oxidative stress is absolutely necessary to enhance the thermotolerance and production of .

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References

Arrigo A, Paul C, Ducasse C, Sauvageot O, Kretz-Remy C . Small stress proteins: modulation of intracellular redox state and protection against oxidative stress. Prog Mol Subcell Biol. 2002; 28:171-84. DOI: 10.1007/978-3-642-56348-5_9. View

Jia H, Fan Y, Feng X, Li C . Enhancing stress-resistance for efficient microbial biotransformations by synthetic biology. Front Bioeng Biotechnol. 2014; 2:44. PMC: 4202804. DOI: 10.3389/fbioe.2014.00044. View

Shao Z, Zhao H, Zhao H . DNA assembler, an in vivo genetic method for rapid construction of biochemical pathways. Nucleic Acids Res. 2008; 37(2):e16. PMC: 2632897. DOI: 10.1093/nar/gkn991. View

Sakaki K, Tashiro K, Kuhara S, Mihara K . Response of genes associated with mitochondrial function to mild heat stress in yeast Saccharomyces cerevisiae. J Biochem. 2003; 134(3):373-84. DOI: 10.1093/jb/mvg155. View

Tomas-Pejo E, Oliva J, Ballesteros M, Olsson L . Comparison of SHF and SSF processes from steam-exploded wheat straw for ethanol production by xylose-fermenting and robust glucose-fermenting Saccharomyces cerevisiae strains. Biotechnol Bioeng. 2008; 100(6):1122-31. DOI: 10.1002/bit.21849. View

Davidson J, Schiestl R . Mitochondrial respiratory electron carriers are involved in oxidative stress during heat stress in Saccharomyces cerevisiae. Mol Cell Biol. 2001; 21(24):8483-9. PMC: 100011. DOI: 10.1128/MCB.21.24.8483-8489.2001. View

Doyle S, Genest O, Wickner S . Protein rescue from aggregates by powerful molecular chaperone machines. Nat Rev Mol Cell Biol. 2013; 14(10):617-29. DOI: 10.1038/nrm3660. View

Jia H, Sun X, Sun H, Li C, Wang Y, Feng X . Intelligent Microbial Heat-Regulating Engine (IMHeRE) for Improved Thermo-Robustness and Efficiency of Bioconversion. ACS Synth Biol. 2016; 5(4):312-20. DOI: 10.1021/acssynbio.5b00158. View

Vandenbroucke K, Robbens S, Vandepoele K, Inze D, Van de Peer Y, Van Breusegem F . Hydrogen peroxide-induced gene expression across kingdoms: a comparative analysis. Mol Biol Evol. 2008; 25(3):507-16. DOI: 10.1093/molbev/msm276. View

10.

Wieser R, Adam G, Wagner A, Schuller C, Marchler G, Ruis H . Heat shock factor-independent heat control of transcription of the CTT1 gene encoding the cytosolic catalase T of Saccharomyces cerevisiae. J Biol Chem. 1991; 266(19):12406-11. View

11.

Richter K, Haslbeck M, Buchner J . The heat shock response: life on the verge of death. Mol Cell. 2010; 40(2):253-66. DOI: 10.1016/j.molcel.2010.10.006. View

12.

Luan G, Dong H, Zhang T, Lin Z, Zhang Y, Li Y . Engineering cellular robustness of microbes by introducing the GroESL chaperonins from extremophilic bacteria. J Biotechnol. 2014; 178:38-40. DOI: 10.1016/j.jbiotec.2014.03.010. View

13.

Egorova K, Antranikian G . Industrial relevance of thermophilic Archaea. Curr Opin Microbiol. 2005; 8(6):649-55. DOI: 10.1016/j.mib.2005.10.015. View

14.

Caspeta L, Chen Y, Ghiaci P, Feizi A, Buskov S, Hallstrom B . Biofuels. Altered sterol composition renders yeast thermotolerant. Science. 2014; 346(6205):75-8. DOI: 10.1126/science.1258137. View

15.

Benjaphokee S, Koedrith P, Auesukaree C, Asvarak T, Sugiyama M, Kaneko Y . CDC19 encoding pyruvate kinase is important for high-temperature tolerance in Saccharomyces cerevisiae. N Biotechnol. 2011; 29(2):166-76. DOI: 10.1016/j.nbt.2011.03.007. View

16.

Vabulas R, Raychaudhuri S, Hayer-Hartl M, Hartl F . Protein folding in the cytoplasm and the heat shock response. Cold Spring Harb Perspect Biol. 2010; 2(12):a004390. PMC: 2982175. DOI: 10.1101/cshperspect.a004390. View

17.

Haslbeck M, Franzmann T, Weinfurtner D, Buchner J . Some like it hot: the structure and function of small heat-shock proteins. Nat Struct Mol Biol. 2005; 12(10):842-6. DOI: 10.1038/nsmb993. View

18.

Dunlop M, Dossani Z, Szmidt H, Chu H, Lee T, Keasling J . Engineering microbial biofuel tolerance and export using efflux pumps. Mol Syst Biol. 2011; 7:487. PMC: 3130554. DOI: 10.1038/msb.2011.21. View

19.

Lin Z, Zhang Y, Wang J . Engineering of transcriptional regulators enhances microbial stress tolerance. Biotechnol Adv. 2013; 31(6):986-91. DOI: 10.1016/j.biotechadv.2013.02.010. View

20.

Liu Y, Zhang G, Sun H, Sun X, Jiang N, Rasool A . Enhanced pathway efficiency of Saccharomyces cerevisiae by introducing thermo-tolerant devices. Bioresour Technol. 2014; 170:38-44. DOI: 10.1016/j.biortech.2014.07.063. View