Using flow cytometry to monitor the stress response of yeast and microalgae populations in mixed cultures developed in brewery effluents

Springer Science and Business Media LLC - Tập 32 - Trang 3687-3701 - 2020
Carla Dias1, Luísa Gouveia1, José A. L. Santos2,3, Alberto Reis1, Teresa Lopes da Silva1
1Laboratório Nacional de Energia e Geologia, I.P., Unidade de Bioenergia e Biorrefinarias, Lisboa, Portugal
2Departamento de Bioengenharia, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal
3IBB, Institute for Biotechnology and Bioengineering, Lisboa, Portugal

Tóm tắt

Recently, yeast and microalgae mixed cultures have been widely used in biological effluent treatments and biofuel production because such cultures show many advantages over pure cultures. However, industrial effluents often contain toxic compounds; therefore, it is important to evaluate the cell stress response when growing in such conditions during the mixed culture development. In this work, flow cytometry (FC) was used to differentiate Rhodosporidium toruloides cells from Tetradesmus obliquus cells, based on their size, internal complexity, and chlorophyll content. FC coupled with SYTOX Green and CFDA fluorochromes was also used to characterize the cell stress response of R. toruloides and T. obliquus individual cells in a mixed culture. This work describes, for the first time, a simple and easy method to monitor individual stress response of R. toruloides and T. obliquus cells growing in mixed cultures on brewery effluents, using FC coupled with fluorescent dyes.

Tài liệu tham khảo

Adan A, Alizada G, Kiraz Y, Baran Y, Nalbant A (2017) Flow cytometry: basic principles and applications. Crit Rev Biotechnol 37:163–176 APHA (1998) Standard Methods for the Examination of Water and Wastewater. 20th Edition, American Public Health Association, American Water Works Association and Water Environmental Federation, Washington, DC Bassey EN, Inyang UE, Inyang JD (2012) Characterization of brewery effluent fluid. J Eng Appl Sci 4:67–77 Borowitzka MA (2018) The ‘stress’ concept in microalgal biology – homeostasis, acclimation and adaptation. J Appl Phycol 30:2815–2825 Cheirsilp B, Kitcha S, Torpee S (2011) Co-culture of an oleaginous yeast Rhodotorula glutinis and a microalga Chlorella vulgaris for biomass and lipid production using pure and crude glycerol as a sole carbon source. Ann Microbiol 62:987–993 Dias C, Sousa S, Caldeira J, Reis A, Lopes da Silva T (2015) New dual-stage pH control fed-batch cultivation strategy for the improvement of lipids and carotenoids production by the red yeast Rhodosporidium toruloides NCYC 921. Bioresour Technol 189:309–318 Dias C, Silva C, Freitas C, Reis A, Lopes da Silva T (2016) Effect of medium pH on Rhodosporidium toruloides NCYC 921 carotenoid and lipid production evaluated by flow cytometry. Appl Biochem Biotech 179:776–787 Dias C, Santos J, Reis A, Lopes da Silva T (2019) Yeast and microalgal symbiotic cultures using low-cost substrates for lipid production. Bioresour Technol Reports 7:100261 Díaz M, Herrero M, García LA, Quirós C (2010) Application of flow cytometry to industrial microbial bioprocesses. Biochem Eng J 48:385–407 El-Sheekh M, Abomohra AE, Hanelt D (2013) Optimization of biomass and fatty acid productivity of Scenedesmus obliquus as a promising microalga for biodiesel production. World J Microbiol Biotechnol 29:915–922 El-Sheekh MM, Bedaiwy MY, Osman ME, Ismail MM (2014) Influence of molasses on growth, biochemical composition and ethanol production of the green algae Chlorella vulgaris and Scenedesmus obliquus. J Agric Eng Biotechnol 2:20–28 Ferreira VS, Pinto RF, Sant’Anna C (2015) Low light intensity and nitrogen starvation modulate the chlorophyll content of Scenedesmus dimorphus. J App Microbiol 120:661–670 Ferreira A, Ribeiro B, Marques PASS, Ferreira AF, Dias AP, Pinheiro HM, Reis A, Gouveia L (2017) Scenedesmus obliquus mediated brewery wastewater remediation and CO2 biofixation for green energy purposes. J Cleaner Prod 165:1316–1327 Freitas C, Nobre B, Gouveia L, Roseiro J, Reis A, Lopes da Silva T (2014a) New at-line flow cytometric protocols for determining carotenoid content and cell viability during Rhodosporidium toruloides NCYC 921 batch growth. Process Biochem 49:554–562 Freitas C, Parreira TM, Roseiro J, Reis A, Lopes da Silva T (2014b) Selecting low-cost carbon sources for carotenoid and lipid production by the pink yeast Rhodosporidium toruloides NCYC 921 using flow cytometry. Bioresour Technol 158:355–359 Guyot S, Gervais P, Young M, Winckler P, Dumont J, Davey HM (2015) Surviving the heat: heterogeneity of response in Saccharomyces cerevisiae provides insight into thermal damage to the membrane. Environ Microbiol 17:2982–2992 Haddad SA, Lindegrena CC (1953) Method for determining the weight of an individual yeast cell. Appl Microbiol 1:153–156 Horwitz W, Latimer GW (2005) Official Methods of Analysis of AOAC International, 18th edition, Association of Official Analytical Chemistry International, Maryland Hu W (2014) Dry weight and cell density of individual algal and cyanobacterial cells for algae research and development. University of Missouri-Columbia, USA, MSc Thesis, 81 pp Jiang X, Liu L, Chen J, Wei D (2018) Effects of Xanthophyllomyces dendrorhous on cell growth, lipid, and astaxanthin production of Chromochloris zofingiensis by mixed culture strategy. J Appl Phycol 30:3009–3015 La A, Perré P, Taidi B (2019) Process for symbiotic culture of Saccharomyces cerevisiae and Chlorella vulgaris for in situ CO2 mitigation. Appl Microbiol Biotechnol 103:731–745 Liu L, Chen J, Lim PE, Wei D (2018a) Enhanced single cell oil production by mixed culture of Chlorella pyrenoidosa and Rhodotorula glutinis using cassava bagasse hydrolysate as carbon source. Bioresour Technol 255:140–148 Liu L, Chen J, Lim PE, Wei D (2018b) Dual-species cultivation of microalgae and yeast for enhanced biomass and microbial lipid production. J Appl Phycol 30:2997–3007 Lopes da Silva T, Roseiro JC, Reis A (2012) Applications and perspectives of multiparameter flow cytometry to microbial biofuels production processes. Trends Biotechnol 30:225–232 Marchão L, Lopes da Silva T, Gouveia L, Reis A (2018) Microalgae-mediated brewery wastewater treatment: effect of dilution rate on nutrient removal rates, biomass biochemical composition, and cell physiology. J Appl Phycol 30:1583–1595 Martins V, Dias C, Caldeira J, Duarte LC, Reis A, Lopes da Silva T (2018) Carob pulp syrup: A potential Mediterranean carbon source for carotenoids production by Rhodosporidium toruloides NCYC 921. Bioresour Technol Reports 3:177–184 Monteiro CM, Castro PM, Malcata FX (2012) Metal uptake by microalgae: underlying mechanisms and practical applications. Biotechnol Prog 28:299–311 Papone T, Kookkhunthod S, Paungbut M, Leesing R (2016) Producing of microbial oil by mixed culture of microalgae and oleaginous yeast using sugarcane molasses as carbon substrate. J Clean Energy Technol 4:1–4 Qin L, Liu L, Wang Z, Chen W, Wei D (2018a) Efficient resource recycling from liquid digestate by microalgae-yeast mixed culture and the assessment of key gene transcription related to nitrogen assimilation in microalgae. Bioresour Technol 264:90–97 Qin L, Wei D, Wang Z, Alam MA (2018b) Advantage assessment of mixed culture of Chlorella vulgaris and Yarrowia lipolytica for treatment of liquid digestate of yeast industry and cogeneration of biofuel feedstock. Appl Biochem Biotechnol 187:856–869 Qin L, Liu L, Wang Z, Chen W, Wei D (2019) The mixed culture of microalgae Chlorella pyrenoidosa and yeast Yarrowia lipolytica for microbial biomass production. Bioprocess Biosyst Eng 42:1409–1419 Rinanti A, Kardena E, Astuti DI, Dewi K (2013) Growth response and chlorophyll content of Scenedesmus obliquus cultivated in different artificial media. Asian J Environ Biol 1:1–9 Santos CA, Caldeira ML, Lopes da Silva T, Novais JM, Reis A (2013) Enhanced lipidic algae biomass production using gas transfer from a fermentative Rhodosporidium toruloides culture to an autotrophic Chlorella protothecoides culture. Bioresour Technol 138:48–54 Shu CH, Tsai CC (2016) Enhancing oil accumulation of a mixed culture of Chlorella sp. and Saccharomyces cerevisiae using fish waste hydrolysate. J Taiwan Inst Chem Eng 67:377–384 Takahashi T (2019) Routine management of microalgae using autofluorescence from chlorophyll. Molecules 24:4441–4456 Wang S, Wu Y, Wang X (2016) Heterotrophic cultivation of Chlorella pyrenoidosa using sucrose as the sole carbon source by co-culture with Rhodotorula glutinis. Bioresour Technol 220:615–620 Wang SK, Xu W, Tao HH, Sun XS, Tian YT (2018) Heterotrophic culture of Chlorella pyrenoidosa using sucrose as the sole carbon source by co-culture with immobilized yeast. Bioresour Technol 249:425–430 Willaert R, De Backer L, Baron GV (1996) Modelling the immobilisation of cells in a packed bed of porous carriers. In: Buitelaar RM, Bucke C, Tramper J, Wijffels RH (eds) Immobilized cells: basics and applications. Elsevier, Amsterdam, pp 154–161 Yoon S, Rhee J (1983) Lipid from yeast fermentation: Effects of cultural conditions on lipid production and its characteristics of Rhodotorula glutinis. J Am Oil Chem Soc 60:1281–1286 Zhang Z, Ji H, Gong G, Zhang X, Tan T (2014) Synergistic effects of oleaginous yeast Rhodotorula glutinis and microalga Chlorella vulgaris for enhancement of biomass and lipid yields. Bioresour Technol 164:93–99 Zhang K, Zheng J, Xue D, Ren D, Lu J (2017) Effect of photoautotrophic and heteroautotrophic conditions on growth and lipid production in Chlorella vulgaris cultured in industrial wastewater with the yeast Rhodotorula glutinis. J Appl Phycol 29:2783–2788