Quantifying the parametric sensitivity of ethanol production by Scheffersomyces (Pichia) stipitis: development and verification of a method based on the principles of growth on mixtures of complementary substrates Free

Abstract

Under aerobic conditions, Crabtree-negative yeasts grow but do not ferment, and under anaerobic conditions, they ferment but do not grow. It is therefore believed that fermentation by these yeasts is sensitive to small variations of the operating parameters, e.g. dilution rate , mass transfer coefficient and oxygen solubility . However, this parametric sensitivity has never been quantified. Here, we present a method to quantify the parametric sensitivity of ethanol production in the Crabtree-negative yeast Scheffersomycesstipitis. The method is based on our experimental observation that S. stipitis cultures follow the principles of growth on mixtures of complementary substrates. Specifically, if a chemostat operating at fixed , and is fed with progressively increasing glucose feed concentrations , the culture passes through three regimes. (1) At low , the culture is carbon-limited and no ethanol is produced. (2) At high , the culture is oxygen-limited and ethanol is produced, but unused glucose is lost with the effluent. (3) At intermediate , both glucose and oxygen are limiting, and ethanol is produced without loss of glucose. Ethanol must therefore be produced in this dual-limited regime. The dual-limited regime can be predicted by simple unstructured models. It is characterized by the relation , where and denote the g of glucose consumed per g of oxygen during carbon- and oxygen-limited growth. Hence, the parametric sensitivity of fermentation by Crabtree-negative yeasts can be improved by targeting the yields and .

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2018-09-28
2024-03-29
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References

  1. Kurtzman CP, Suzuki M. Phylogenetic analysis of ascomycete yeasts that form coenzyme Q-9 and the proposal of the new genera Babjeviella, Meyerozyma, Millerozyma, Priceomyces, and Scheffersomyces. Mycoscience 2010; 51:2–14 [View Article]
    [Google Scholar]
  2. Toivola A, Yarrow D, van den Bosch E, van Dijken JP, Scheffers WA. Alcoholic fermentation of d-xylose by yeasts. Appl Environ Microbiol 1984; 47:1221–1223[PubMed]
    [Google Scholar]
  3. du Preez JC, Prior BA. A quantitative screening of some xylose-fermenting yeast isolates. Biotechnol Lett 1985; 7:241–246 [View Article]
    [Google Scholar]
  4. Delgenes JP, Moletta R, Navarro JM. The effect of aeration on D-xylose fermentation by Pachysolen tannophilus, Pichia stipitis, Kluyveromyces marxianus and Candida shehatae. Biotechnol Lett 1986; 8:897–900 [View Article]
    [Google Scholar]
  5. Dellweg H, Rizzi M, Klein C. Controlled limited aeration and metabolic regulation during the production of ethanol from D-xylose by Pichia stipitis. J Biotechnol 1989; 12:111–122 [View Article]
    [Google Scholar]
  6. Grootjen DRJ, van der Lans R, Luyben K. Conversion of glucose/xylose mixtures by Pichia stipitis under oxygen-limited conditions. Enzyme Microb Technol 1991; 13:648–654 [View Article]
    [Google Scholar]
  7. du Preez JC, van Driessel B, Prior BA. Ethanol tolerance of Pichia stipitis and Candida shehatae strains in fed-batch cultures at controlled low dissolved oxygen levels. Appl Microbiol Biotechnol 1989; 30:53–58 [View Article]
    [Google Scholar]
  8. van Dijken JP, Weusthuis RA, Pronk JT. Kinetics of growth and sugar consumption in yeasts. Antonie van Leeuwenhoek 1993; 63:343–352 [View Article][PubMed]
    [Google Scholar]
  9. Egli T. On multiple-nutrient-limited growth of microorganisms, with special reference to dual limitation by carbon and nitrogen substrates. Antonie van Leeuwenhoek 1991; 60:225–234 [View Article][PubMed]
    [Google Scholar]
  10. Durner R, Witholt B, Egli T. Accumulation of Poly[(R)-3-hydroxyalkanoates] in Pseudomonas oleovorans during growth with octanoate in continuous culture at different dilution rates. Appl Environ Microbiol 2000; 66:3408–3414 [View Article][PubMed]
    [Google Scholar]
  11. Bandyopadhyay B, Humphrey AE, Taguchi H. Dynamic measurement of the volumetric oxygen transfer coefficient in fermentation systems. Biotechnol Bioeng 1967; 9:533–544 [View Article]
    [Google Scholar]
  12. Brooks JD, MacLennan DG, Barford JP, Hall RJ. Design of laboratory continuous-culture equipment for accurate gaseous metabolism measurements. Biotechnol Bioeng 1982; 24:847–856 [View Article][PubMed]
    [Google Scholar]
  13. Egli T, Fiechter A. Theoretical analysis of media used in the growth of yeasts on methanol. Microbiology 1981; 123:365–369 [View Article]
    [Google Scholar]
  14. du Preez JC, Bosch M, Prior BA. The fermentation of hexose and pentose sugars by Candida shehatae and Pichia stipitis. Appl Microbiol Biotechnol 1986; 23:228–233 [View Article]
    [Google Scholar]
  15. Skoog K, Jeppsson H, Hahn-Hägerdal B. The effect of oxygenation on glucose fermentation with pichia stipitis. Appl Biochem Biotechnol 1992; 34-35:369–375 [View Article]
    [Google Scholar]
  16. Skoog K, Hahn-Hägerdal B. Effect of oxygenation on xylose fermentation by Pichia stipitis. Appl Environ Microbiol 1990; 56:3389–3394[PubMed]
    [Google Scholar]
  17. Grootjen DRJ, van der Lans R, Luyben K. Effects of the aeration rate on the fermentation of glucose and xylose by Pichia stipitis CBS 5773. Enzyme Microb Technol 1990; 12:20–23 [View Article]
    [Google Scholar]
  18. du Preez JC. Process parameters and environmental factors affecting d-xylose fermentation by yeasts. Enzyme Microb Technol 1994; 16:944–956 [View Article]
    [Google Scholar]
  19. Duboc P, von Stockar U. Systematic errors in data evaluation due to ethanol stripping and water vaporization. Biotechnol Bioeng 1998; 58:428–439 [View Article][PubMed]
    [Google Scholar]
  20. Wyman CE. What is (and is not) vital to advancing cellulosic ethanol. Trends Biotechnol 2007; 25:153–157 [View Article][PubMed]
    [Google Scholar]
  21. Danckwerts PV. Continuous flow systems: Distribution of residence time. Chemical Engineering Science 1953; 2:1–13
    [Google Scholar]
  22. Garcia-Ochoa F, Gomez E. Bioreactor scale-up and oxygen transfer rate in microbial processes: an overview. Biotechnol Adv 2009; 27:153–176 [View Article][PubMed]
    [Google Scholar]
  23. Slininger PJ, Branstrator LE, Bothast RJ, Okos MR, Ladisch MR. Growth, death, and oxygen uptake kinetics of Pichia stipitis on xylose. Biotechnol Bioeng 1991; 37:973–980 [View Article][PubMed]
    [Google Scholar]
  24. Alexander MA, Chapman TW, Jeffries TW. Continuous ethanol production from D-xylose by Candida shehatae. Biotechnol Bioeng 1987; 30:685–691 [View Article][PubMed]
    [Google Scholar]
  25. von Stockar U, Birou B. The heat generated by yeast cultures with a mixed metabolism in the transition between respiration and fermentation. Biotechnol Bioeng 1989; 34:86–101 [View Article][PubMed]
    [Google Scholar]
  26. von Stockar U, Auberson LC. Chemostat cultures of yeasts, continuous culture fundamentals and simple unstructured mathematical models. J Biotechnol 1992; 22:69–87 [View Article][PubMed]
    [Google Scholar]
  27. Weusthuis RA, Visser W, Pronk JT, Scheffers WA, van Dijken JP. Effects of oxygen limitation on sugar metabolism in yeasts: a continuous-culture study of the Kluyver effect. Microbiology 1994; 140:703–715 [View Article][PubMed]
    [Google Scholar]
  28. Egli T, Zinn M. The concept of multiple-nutrient-limited growth of microorganisms and its application in biotechnological processes. Biotechnol Adv 2003; 22:35–43 [View Article][PubMed]
    [Google Scholar]
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