Abstract
Ethanol-tolerant Arthrobacter simplex is desirable since ethanol facilitates hydrophobic substrates dissolution on an industrial scale. Herein, alterations in compatible solutes were investigated under ethanol stress. The results showed that the amount of trehalose and glycerol increased while that of glutamate and proline decreased. The trehalose protectant role was verified and its concentration was positively related to the degree of cell tolerance. otsA, otsB and treS, three trehalose biosynthesis genes in A. simplex, also enhanced Escherichia coli stress tolerance, but the increased tolerance was dependent on the type and level of the stress. A. simplex strains accumulating trehalose showed a higher productivity in systems containing more ethanol and substrate because of better viability. The underlying mechanisms of trehalose were involved in better cell integrity, higher membrane stability, stronger reactive oxygen species scavenging capacity and higher energy level. Therefore, trehalose was a general protectant and the upregulation of its biosynthesis by genetic modification enhanced cell stress tolerance, consequently promoted productivity.
Similar content being viewed by others
References
Song B, Zhou Q, Xue HJ, Liu JJ, Zheng YY, Shen YB, Wang M, Luo JM (2018) IrrE improves organic solvent tolerance and Δ1-dehydrogenation productivity of Arthrobacter simplex. J Agric Food Chem 66:5210–5220
Vianna CR, Silva CLC, Neves MJ, Rosa CA (2008) Saccharomyce scerevisiae strains from traditional fermentations of Brazilian cachaça: trehalose metabolism, heat and ethanol resistance. Antonie Van Leeuwenhoek 93:205–217
Nicolaou SA, Gaida SM, Papoutsakis ETA (2010) A comparative view of metabolite and substrate stress and tolerance in microbial bioprocessing: from biofuels and chemicals, to biocatalysis and bioremediation. Metab Eng 12(4):307–331
Mahmud SA, Hirasawa T, Shimizu H (2010) Differential importance of trehalose accumulation in Saccharomyces cerevisiae in response to various environmental stresses. J Biosci Bioeng 109(3):262–266
Mansure JJC, Panek A, Crowe LM, Crowe JH (1994) Trehalose inhibits ethanol effects on intact yeast cells and liposomes. Biochim Biophys Acta 1191(2):309–316
Mansure JJ, Souza RC, Panek AD (1997) Trehalose metabolism in Saccharomyces cerevisiae during alcoholic fermentation. Biotechnol Lett 19:1201–1203
Ogawa Y, Nitta A, Uchiyama H, Imamura T, Shimoi H, Ito K (2000) Tolerance mechanism of the ethanol-tolerant mutant of sake yeast. J Biosci Bioeng 90(3):313–320
Pataro C, Guerra JB, Gomes FCO, Neves MJ, Pimentel PF, Rosa CA (2002) Trehalose accumulation, invertase activity and physiological characteristics of yeasts isolated from 24 h fermentative cycles during the production of artisanal Brazilian cachaça. Braz. J Microbiol 33:202–208
Sharma SC (1997) A possible role of trehalose in osmotolerance and ethanol tolerance in Saccharomyces cerevisiae. FEMS Microbiol Lett 152:11–15
Van Laere A (1989) Trehalose, reserve and/or stress metabolite? FEMS Microbiol Rev 63:201–210
Wiemken A (1990) Trehalose in yeast, stress protectant rather than reserve carbohydrate. Antonie Leeuwenhoek 58:209–217
Bandara A, Fraser S, Chambers PJ, Stanley GA (2009) Trehalose promotes the survival of Saccharomyces cerevisiae during lethal ethanol stress, but does not influence growth under sublethal ethanol stress. FEMS Yeast Res 9(8):1208–1216
Alexandre H, Plourde L, Charpentier C, Francois J (1998) Lack of correlation between trehalose accumulation, cell viability and intracellular acidification as induced by various stresses in Saccharomyces cerevisiae. Microbiology 144:1103–1111
Gomes FCO, Pataro C, Guerra JB, Neves MJ, Correa SR (2002) Physiological diversity and trehalose accumulation in Schizosaccharomyces pombe strains isolated from spontaneous fermentations during the production of the artisanal Brazilian cachaça. Can J Microbiol 48:399–406
Ribeiro MJS, Leao LSC, Morais PB, Rosa CA, Panek AD (1999) Trehalose accumulation by tropical yeast strains submitted to stress conditions. Antonie Van Leeuwenhoek 75:245–251
Kim J, Alizadeh P, Harding T, Hefner-Gravink A, Klionsky DJ (1996) Disruption of the yeast ATH1 gene confers better survival after dehydration, freezing, and ethanol shock: Potential commercial applications. Appl Environ Microbiol 62(5):1563–1569
Jung YJ, Park HD (2005) Antisense-mediated inhibition of acid trehalase (ATH1) gene expression promotes ethanol fermentation and tolerance in Saccharomyces cerevisiae. Biotechnol Lett 27(23–24):1855–1859
Soto T, Fernandez J, Vicente-Soler J, Cansado J, Gacto M (1999) Accumulation of trehalose by overexpression of tps1, coding for trehalose-6-phosphate synthase, causes increased resistance to multiple stresses in the fission yeast Schizosaccharomyces pombe. Appl Environ Microbiol 65(5):2020–2024
Takagi H, Takaoka M, Kawaguchi A, Kubo Y (2005) Effect of L-proline on sake brewing and ethanol stress in Saccharomyces cerevisiae. Appl Environ Microbiol 71:8656–8662
Wang PM, Zheng DQ, Chi XQ, Li O, Qian CD, Liu TZ, Zhang XY, Du FG, Sun PY, Qu AM, Wu XC (2014) Relationship of trehalose accumulation with ethanol fermentation in industrial Saccharomyces cerevisiae yeast strains. Bioresource Technol 152:371–376
Woodruff L, Boyle NR, Gill RT (2013) Engineering improved ethanol production in Escherichia coli with a genome-wide approach. Metab Eng 17:1–11
Thomas KC, Hynes SH, Ingledew WM (1994) Effects of particulate materials and osmoprotectants on very-high-gravity ethanolic fermentation by Saccharomyces cerevisiae. Appl Environ Microbiol 60:1519–1524
Gonzalez R, Tao H, Purvis JE, York SW, Shanmugam KT, Ingram LO (2003) Gene array-based identification of changes that contribute to ethanol tolerance in ethanologenic Escherichia coli: comparison of KO11 (parent) to LY01 (resistant mutant). Biotechnol Prog 19:612–623
Underwood SA, Buszko ML, Shanmugam KT, Ingram LO (2004) Lack of protective osmolytes limits final cell density and volumetric productivity of ethanologenic Escherichia coli KO11 during xylose fermentation. Appl Environ Microbiol 70:2734–2740
Luo JM, Song ZY, Ning J, Cheng YX, Wang YX, Cui FF, Shen Y, Wang M (2018) The global ethanol-induced alteration in Arthrobacter simplex and its mutants with enhanced ethanol tolerance. Appl Microbiol Biotechnol 102:9331–9350
Sandu C, Chiribau CB, Sachelaru P, Brandsch R (2005) Plasmids for nicotine-dependent and -independent gene expression in Arthrobacter nicotinovorans and other Arthrobacter species. Appl Environ Microbiol 71:8920–8924
Luo JM, Ning J, Wang YX, Cheng YX, Zheng Y, Shen YB, Wang M (2013) The effect of ethanol on cell properties and steroid 1-en-dehydrogenation biotransformation of Arthrobacter simplex. Biotechnol Appl Bioc 61:555–564
Kaino T, Takagi H (2008) Gene expression profiles and intracellular contents of stress protectants in Saccharomyces cerevisiae under ethanol and sorbitol stresses. Appl Microbiol Biotechnol 79:273–283
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408
Devantier R, Scheithauer B, Villas-Boas SG, Pedersen S, Olsson L (2005) Metabolite profiling for analysis of yeast stress response during very high gravity ethanol fermentations. Biotechnol Bioeng 90:703–714
Liang LK, Wang XK, Zhu KL, Chi ZM (2007) Trehalose synthesis in Saccharomycopsis fibuligera does not respond to stress treatment. Appl Microbiol Biotechnol 74:1084–1091
Nagahisa K, Hirasawa T, Mahmud AS, Shimizu H, Yoshikawa K, Ashitani K (2010) Effect of trehalose accumulation on response to saline stress in Saccharomyces cerevisiae. Yeats 26:17–30
Vilchez S, Tunnacliffe A, Manzanera M (2008) Tolerance of plastic-encapsulated Pseudomonas putida KT2440 to chemical stress. Extremophiles 12(2):297–299
Manzanera M, Vilchez S, Tunnacliffe A (2004) Plastic encapsulation of stabilized Escherichia coli and Pseudomonas putida. Appl Environ Microbiol 70(5):3143–3145
Purvis JE, Yomano LP, Ingram LO (2005) Enhanced trehalose production improves growth of Escherichia coli under osmotic stress. Appl Environ Microbiol 71:3761–3769
Miller EN, Ingram LO (2008) Sucrose and overexpression of trehalose biosynthetic genes (otsBA) increase desiccation tolerance of recombinant Escherichia coli. Biotechnol Lett 30:503–508
Nguyen AQ, Kim YG, Kim SB, Kim CJ (2013) Improved tolerance of recombinant Escherichia coli to the toxicity of crude glycerol by overexpressing trehalose biosynthetic genes (otsBA) for the production of β-carotene. Bioresource Technol 143:531–537
Ingram LO (1986) Microbial tolerance to alcohol:Role of cell membrane. Trends Biotechnol 4:40–44
Barry JA, Gawrisch K (1995) Effects of ethanol on lipid bilayers containing cholesterol, gangliosides and sphingomyelin. Biochemistry 34:8852–8860
Crowe JH, Crowe LM, Chapman D (1984) Preservation of membranes in anhydrobiotic organisms: the role of trehalose. Science 223:701–703
Iwahashi H, Obachi K, Fujii S, Komatsu Y (1995) The correlative evidence suggesting that trehalose stabilizes membrane structure in the yeast Saccharomyces cerevisiae. Cell Mol Biol 41:763–766
Crowe JH, Carpenter JF, Crowe LM (1998) The role of vitrification in anhydrobiosis. Annu Rev Physiol 60:73–103
Arguelles JC (2000) Physiological roles of trehalose in bacteria and yeasts: a comparative analysis. Arch Microbiol 174:217–224
Singer MA, Lindquist S (1998) Multiple effects of trehalose on protein folding in vitro and in vivo. Mol Cell 1:639–648
Alvarez-Peral FJ, Zaragoza O, Pedreno Y, Arguelles JC (2002) Protective role of trehalose during severe oxidative stress caused by hydrogen peroxide and the adaptive oxidative stress response in Candida albicans. Microbiology 148:2599–2606
Herdeiro RS, Pereira MD, Panek AD, Eleutherio ECA (2006) Trehalose protects Saccharomyces cerevisiae from lipid peroxidation during oxidative stress. Biochim Biophys Act 1760:340–346
Nery DCM, Silva CG, Mariani D, Fernandes PN, Pereira MD, Panek AD, Eleutherio ECA (2008) The role of trehalose and its transporter in protection against reactive oxygen species. Biochim Biophys Acta 1780:1408–1411
Bowles LK, Ellefson WL (1985) Effects of butanol on Clostridium acetobutylicum. Appl Environ Microbiol 50(5):1165–1170
Heipieper HJ, Keweloh H, Rehm HJ (1991) Influence of phenols on growth and membrane permeability of free and immobilized Escherichia coli. Appl Environ Microbiol 57:1213–1217
Acknowledgements
This work was financially supported by Natural Science Foundation of China (nos. 21978220), the Natural Science Foundation of Tianjin (nos. 18JCZDJC32500), Tianjin Technical Expert Project (nos. 19JCTPJC50800), the Open Fund of Ministry of Education Key Laboratory of Molecular Microbiology and Technology, Nankai University, Tianjin Municipal Science and Technology Commission (17PTGCCX00190), and 2019 Tianjin innovation and entrepreneurship training program for college students (national grade) (201910057051).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Cheng, Hj., Sun, Yh., Chang, Hw. et al. Compatible solutes adaptive alterations in Arthrobacter simplex during exposure to ethanol, and the effect of trehalose on the stress resistance and biotransformation performance. Bioprocess Biosyst Eng 43, 895–908 (2020). https://doi.org/10.1007/s00449-020-02286-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-020-02286-9