A Simple Metabolic Flux Balance Analysis of Biomass and Bioethanol Production in Kluyveromyces Marxianus ATCC 26548 Batch Culture


  • Mohamad Fahrurrazi Tompang
  • Khairiah Abd Karim
  • Azlina Harun @Kamaruddin
  • Ku Syahidah Ku Ismail
  • . .
  • . .






Metabolic flux balance, Kluyveromyces marxianus, oxidative metabolism, reductive metabolism, oxygen uptake rate


The role of a non-conventional yeast, Kluyveromyces marxianus in the bioprocessing industry has shown potential as metabolites producer, making it a suitable candidate for replacing the baker’s yeast for various industrial applications. The mathematical approach is used to analyze the flow of metabolites in the biological system in order to improve the desired product yield as well as the overall production process. Thus, the development of a simple model could lead to sustainability and practicability of the process. In this study, the comparative analysis of a simple metabolic network and a black box description is carried out in order to evaluate the growth and bioethanol production in K. marxianus batch culture. Metabolic flux balance methodology has shown to give a more accurate estimation with the complete analysis of the reaction rates. Furthermore, better evaluation of yeast behavior and performance in a batch system at varying glucose concentrations were achieved based on its stoichiometric reaction analysis. At the highest substrate concentration used, biomass growth was maximum at 12.32 g/l, with 7.75 g/L ethanol obtained. The biomass and bioethanol productions were mostly dependent on oxidative and reductive catabolism, respectively, in which the glucose and oxygen uptake rates played the main role in the regulation of the central metabolic networks. Therefore, biomass and ethanol production are strongly reliant on the cellular functionality of yeast in the culture, which shows the superiority of this method over the black box approach.



[1] Pentjuss A, Stalidzans E, Liepins J, Kokina A, Martynova J, Zikmanis&Motzga I (2017), Model-based biotechnological potential analysis of Kluyvromyces marxianus central metabolism. Journal of Industrial Microbiology and Biotechnology 44 (8), 1177-1190.

[2] Cannizzaro C, Valentinotti S & von Stockar U (2004), Control of yeast fed-batch process through regulation of extracellular ethanol concentration. Bioprocess and Biosystems Engineering 26, 377-383.

[3] Wong WC, Song, HS, Lee JH &Ramkrishna D (2010), Hybrid cybernetic model-based simulation of continuous production of lignocellulosic ethanol: Rejecting abruptly changing feed conditions. Control Engineering Practice 18, 177-189.

[4] Wu, WH., Wang FS & Chang MS (2008), Dynamic sensitivity analysis of biological systems. BMC Bioinformatics 9, 1-17.

[5] Kalnenieks U, Pentjuss A, Rutkis R, Stalidzans E & Fell DA (2014), Modeling Zymomonas mobilis Central Metabolism for Novel Metabolic Engineering Strategies. Frontiers in Microbiology 5, 1-7.

[6] Kerkhoven EJ, Lahtvee PJ & Nielsen J (2015), Application of Computational Modeling in Metabolic Engineering of Yeast. FEMS Yeast Research 15(1), 1-13.

[7] Pricen ND, Reed JL & Palsson BO (2004), Genome-scale models of microbial cells: Evaluating the consequences of constraints. Nature Reviews Microbiology 2, 886-897.

[8] Mahadevan R, Burgard AP, Famili I, van Dien S & Schilling CH (2005), Applications of metabolic modelling to drive process development for the production of value-added chemicals. Biotechnology and Bioprococess Engineering 10, 408-417.

[9] Hjersted JL, Henson MA & Mahadevan R (2007), Genome-scale analysis of Saccharomyces cerevisiae metabolism and ethanol production in fed-batch culture. Biotechnology and Bioengineering97, 1190-1204.

[10] Bro CH, Regenberg B, Förster J, & Nielsen J (2006), In silico-aided metabolic engineering of Saccharomyces cerevisiae for improved bioethanol production. Metabolic Engineering 8, 102-111.

[11] Shiyota S, Shimizu H, Hirasawa T, Nagahisa K, Furusawa C, Pandey G& Katakura Y (2007), Metabolic pathway recruiting through genome data analysis for industrial application of Saccharomyces cerevisiae. Biochemical Engineering Journal 36, 28-37.

[12] Burgard AP, Vaidyarama SN &Maranas CD (2001), Minimal reaction sets for Eschericha coli metabolism under different growth requirements and uptake environments. Biotechnology Progress 17, 791-797.

[13] Aranda-Barradas JS, Garibay-Orijel C, Badillo-Corona JA & Salgado-Manjarrez E (2010), A stochiometric analysis of biological xylitol production. Biochemical Engineering Journal 50, 1-9.

[14] Hong SJ, Kim HJ, Kim JW, Lee DH &Seo JH (2015), Optimizing promoters and secretory signal sequences for producing ethanol from inulin by recombinant Saccharomyces cerevisiae carrying Kluyveromycesmarxianus inulinase. Bioprocess and Biosystems Engineering 38, 263–272.

[15] Kim JS, Park JB, Jang SW &Ha SJ (2015), Enhanced xylitol production by mutant Kluyveromyces marxianus 36907-FMEL1 due to improved xylose reductase activity. Applied Biochemistry and Biotechnology176, 1975–1984.

[16] Kim TY, Lee SW &Oh MK (2014), Biosynthesis of 2-phenylethanol from glucose with genetically engineered Kluyveromyces marxianus. Enzyme and Microbial Technology 61–62, 44–47.

[17] Zhang B, Zhang J, Wang D,Han R, Ding R, Sun L &Hong J (2016), Simultaneous fermentation of glucose and xylose at elevated temperatures co-produces ethanol and xylitol through overexpression of a xylose-specific transporter in engineered Kluyveromyces marxianus. Bioresource Technology 216, 227–237.

[18] Zhang J, Zhang B, Wang D, Gao X, Sun L &Hong J (2015), Rapid ethanol production at elevated temperatures by engineered thermotolerantKluyveromycesmarxianus via the NADP(H)-preferring xylose reductase-xylitol dehydrogenase pathway. Metabolic Engineering 31,140– 152.

[19] Verduyn C, Postma E, Scheffers WA &vanDijken JP (1992), Effect of benzoic acid on metabolic fluxes in yeasts: a continuous-culture study on the regulation of respiration and alcoholic fermentation. Yeast 8, 501–517

[20] Nielsen J &Villadsen J, Bioreaction Engineering Principles, Plenum Press, NY, USA, (1994), pp. 100-102

[21] Kiers J, Zeeman AM, Luttik M, Thiele C, Castrillo JI, Steensma HY, van Dijken, JP &Pronk JT (1998), Regulation of alcoholic fermentation in batch and chemostat cultures of Kluyveromyces lactis CBS 2359. Yeast 14, 459–469.

[22] Gancedo JM (1998), Yeast carbon catabolite repression. Microbiology and Molecular Biology Reviews 62, 334-361.

[23] Simeonidis E, Murabito E, Smallbone K & V. Westerhoff H (2010), Why does yeast ferment? A flux balance analysis study. Biochem. Soc. Trans., 1225-1229

[24] Signori L, Passolunghi S, Ruohonen L, Porro D &Branduardi P (2014), Effect of oxygenation and temperature on glucose-xylose fermentation in Kluyveromyces marxinaus CBS712 strain. Microbial Cell Factories 13, 51-63.

[25] Gonzalez-Siso MI, Garcia-Leiro A, Tarrio N& Esperanza CM (2009), Sugar metabolism, redox balance and oxidative stress response in the respiratory yeast Kluyveromyces lactis. Microbial Cell Factories 8, 46-62.

[26] Saliola M, Tramonti A, Lanini C, Cialfi S, De Biase D & Falcone C, (2012) Intracellular NADPH levels affect the oligomeric state of the glucose 6-phosphatedehydrogenase. Eukaryotic Cell 11, 1503–1511.

[27] Fonseca GG, de Carvalho NMB &Gombert AK (2013) Growth of the yeast Kluyveromycesmarxianus CBS 6556 on different sugar combinations as sole carbon and energy source. Applied Microbiology and Biotechnology 97:5055–5067.

[28] Barrera-Martinez I, Gonzalez-Garcia RA, Salgado-Manjarrez E& Aranda-Barradas, JS (2011), A Simple Metabolic Flux Balance Analysis of Biomass and Bioethanol Production in Saccharomyces cerevisae Fed-batch Culture. Biotechnology and Bioprocess Engineering 16,13-22.

View Full Article:

How to Cite

Fahrurrazi Tompang, M., Abd Karim, K., Harun @Kamaruddin, A., Syahidah Ku Ismail, K., ., ., & ., . (2018). A Simple Metabolic Flux Balance Analysis of Biomass and Bioethanol Production in Kluyveromyces Marxianus ATCC 26548 Batch Culture. International Journal of Engineering & Technology, 7(4.40), 5–10. https://doi.org/10.14419/ijet.v7i4.40.24012
Received 2018-12-15
Accepted 2018-12-15
Published 2018-12-16