Skip to main content

Advertisement

Log in

Enhanced α-ketoglutaric acid production and recovery in Yarrowia lipolytica yeast by effective pH controlling

  • Applied microbial and cell physiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

The replacement of chemical synthesis by environmentally friendly energy-efficient technologies for production of valuable metabolites is a principal strategy of developing biotechnological industry all over the world. In the present study, we develop a method for α-ketoglutaric acid (KGA) production from rapeseed oil with the use of Yarrowia lipolytica yeast. Sixty strains of Y. lipolytica yeasts were tested for their ability to produce KGA, and the strain Y. lipolytica 212 (Y. lipolytica VKM Y-2412) was selected as a promising KGA producer. Using a three-stage pH controlling, in which pH was 4.5 in the growth phase, then since 72 to 144 h, pH was maintained at 3.5 and in the later phase of acid production, the titration by KOH was switch off, selected strain produced 106.5 g l−1 of KGA with mass yield of 0.95 g g−1. KGA in the form of monopotassium salt was isolated from the culture broth and purified. The isolation procedure involved separation of biomass, extraction of residual triglycerides, filtrate bleaching, and acidification with mineral acid (to pH 2.8–3.4), concentration, precipitation of mineral salts, and crystallization of the product. The purity of KGA isolated from the culture filtrate reached 99.1 %.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Ali R, Mittal G, Sultana S, Bhatnagar A (2012) Ameliorative potential of alpha ketoglutaric acid (AKG) on acute lung injuries induced by ammonia inhalation in rats. Exp Lung Res 38:435–444

    Article  PubMed  CAS  Google Scholar 

  • Aurich A, Specht R, Müller RA, Stottmeister U, Yovkova V, Otto C, Holz M, Barth G, Heretsch P, Thomas FA, Sicker D, Giannis A (2012) Microbiologically produced carboxylic acids used as building blocks in organic synthesis. In: Wang X, Chen J, Quinn P (eds) Reprogramming Microbial Metabolic Pathways. Springer-Dortrecht-Heidelberg, New York, pp 391–424

    Chapter  Google Scholar 

  • Barrett DG, Yousaf M (2008) Poly(triol α-ketoglutarate) as biodegradable, chemoselective, and mechanically tunable elastomers. Macromolecules 41:6347–6352

    Article  CAS  Google Scholar 

  • Barth G, Gaillardin C (1996) Yarrowia lipolytica. In: Wolf K (ed) Non conventional yeasts in biotechnology: a handbook. Springer, Berlin, pp 313–388

    Chapter  Google Scholar 

  • Burkholder P, McVeigh J, Moyer D (1944) Studies on some growth factors of yeasts. J Bacteriol 48:385–391

    PubMed  CAS  Google Scholar 

  • Campbell B, Roberts M, Kerksick C, Wilborn C, Marcello B, Taylor L, Nassar E, Leutholtz B, Bowden R, Rasmussen C, Greenwood M, Kreider R (2006) Pharmacokinetics, safety, and effects on exercise performance of l-arginine aketoglutarate in trained adult men. Nutrition 22:872–881

    Article  PubMed  CAS  Google Scholar 

  • Chernyavskaya OG, Shishkanova NV, Il’chenko AP, Finogenova TV (2000) Synthesis of α-ketoglutaric acid by Yarrowia lipolytica yeast grown on ethanol. Appl Microbiol Biotechnol 53:152–158

    Article  PubMed  CAS  Google Scholar 

  • Dedyukhina EG, Chistyakova TI, Kamzolova SV, Vinter MV, Vainshtein MB (2012) Arachidonic acid synthesis by glycerol-grown Mortierella alpine. Eur J Lipid Sci Technol 114:833–841

    Article  CAS  Google Scholar 

  • Ermakova IT, Müller P, Finogenova TV, Lozinov AB (1979) Kinetics of Candida lipolytica yeast growth and biosynthesis of alpha-keto acids with thiamine deficiency in media with different carbon sources. Microbiologiia 48:849–853

    CAS  Google Scholar 

  • Finogenova TV, Morgunov IG, Kamzolova SV, Chernyavskaya OG (2005) Organic acid production by the yeast Yarrowia lipolytica: a review of prospects. Appl Biochem Microbiol 41:418–425

    Article  CAS  Google Scholar 

  • Förster A, Jacobs K, Juretzek J, Mauersberger S, Barth G (2007) Overexpression of the ICL1 gene changes the product ration of citric acid production by Yarrowia lipolytica. Appl Microbiol Biotechnol 77(4):681–689

    Article  Google Scholar 

  • Good DW, Droniuk R, Lawford RG, Fein JE (1985) Isolation and characterization of a Saccharomycopsis lipolytica mutant showing increased production of citric acid from canola oil. Can J Microbiol 31:436–440

    Article  CAS  Google Scholar 

  • Hatti-Kaul R, Törnvall U, Gustafsson L, Börjesson P (2007) Industrial biotechnology for the production of bio-based chemicals—a cradle-to-grave perspective. Trends Biotechnol 25:119–124

    Article  PubMed  CAS  Google Scholar 

  • Heretsch P, Thomas F, Aurich A, Krautscheid H, Sicker D, Giannis A (2008) Syntheses with a chiral building block from the citric acid cycle: (2R.3S)-isocitric acid by fermentation of sunflower oil. Angew Chem Int Ed Engl 47:1958–1960

    Article  PubMed  CAS  Google Scholar 

  • Holz M, Otto C, Kretzschmar A, Yovkova V, Aurich A, Pötter M, Marx A, Barth G (2011) Overexpression of alpha-ketoglutarate dehydrogenase in Yarrowia lipolytica and its effect on production of organic acids. Appl Microbiol Biotechnol 89:1519–1526

    Article  PubMed  CAS  Google Scholar 

  • Kamzolova SV, Morgunov IG (2013) α-Ketoglutaric acid production from rapeseed oil by Yarrowia lipolytica yeast. Appl Microbiol Biotechnol. doi:10.1007/s00253-013-4772-6

    Google Scholar 

  • Kamzolova SV, Morgunov IG, Aurich A, Perevoznikova OA, Shishkanova NV, Stottmeister U, Finogenova TV (2005) Lipase secretion and citric acid production in Yarrowia lipolytica yeast grown on animal and vegetable fat. Food Technol Biotechnol 43:113–122

    Google Scholar 

  • Kamzolova SV, Finogenova TV, Morgunov IG (2008) Microbiological production of citric and isocitric acids from sunflower oil. Food Technol Biotechnol 46:51–59

    CAS  Google Scholar 

  • Kamzolova SV, Lunina JN, Morgunov IG (2011) Biochemistry of citric acid production from rapeseed oil by Yarrowia lipolytica yeast. JAOCS 88:1965–1976

    Article  CAS  Google Scholar 

  • Kamzolova SV, Chiglintseva MN, Lunina JN, Morgunov IG (2012) α-Ketoglutaric acid production by Yarrowia lipolytica and its regulation. Appl Microbiol Biotechnol 96:783–791

    Article  PubMed  CAS  Google Scholar 

  • Lozinov AB, Finogenova TV, Glazunova LM, Illarionova VI (1974) Growth limitation in Candida lipolytica cultures and supersynthesis of metabolites. Mikrobiologiia 43:786–790

    PubMed  CAS  Google Scholar 

  • Maldonado P, Desmarquest JP, Gaillardin C, Binet D (1973) Process for getting diploid Candida lipolytica strains for alpha-ketoglutarate fermentation. Patent US 3930946

  • Mathangi DC, Shyamala R, Vijayashree R, Rao KR, Ruckmani A, Vijayaraghavan R, Bhattacharya R (2011) Effect of alpha-ketoglutarate on neurobehavioral, neurochemical and oxidative changes caused by sub-chronic cyanide poisoning in rats. Neurochem Res 36:540–548

    Article  PubMed  CAS  Google Scholar 

  • Matzi V, Lindenmann J, Muench A, Greilberger J, Juan H, Wintersteiger R, Maier A, Smolle-Juettner FM (2007) The impact of preoperative micronutrient supplementation in lung surgery. A prospective randomized trial of oral supplementation of combined alpha-ketoglutaric acid and 5-hydroxymethylfurfural. Eur J Cardiothorac Surg 32:776–782

    Article  PubMed  Google Scholar 

  • Morgunov IG, Kamzolova SV, Perevoznikova OA, Shishkanova NV, Finogenova TV (2004) Pyruvic acid production by a thiamine auxotroph of Yarrowia lipolytica. Process Biochem 39:1469–1474

    Article  CAS  Google Scholar 

  • Otto C, Yovkova V, Barth G (2011) Overproduction and secretion of α-ketoglutaric acid by microorganisms. Appl Microbiol Biotechnol 92:689–695

    Article  PubMed  CAS  Google Scholar 

  • Otto C, Yovkova V, Aurich A, Mauersberger S, Barth G (2012) Variation of the by-product spectrum during α-ketoglutaric acid production from raw glycerol by overexpression of fumarase and pyruvate carboxylase genes in Yarrowia lipolytica. Appl Microbiol Biotechnol 95:905–917

    Article  PubMed  CAS  Google Scholar 

  • Papanikolaou S, Aggelis G (2002) Lipid production by Yarrowia lipolytica growing on industrial glycerol in a single-stage continuous culture. Bioresour Technol 82:43–49

    Article  PubMed  CAS  Google Scholar 

  • Papanikolaou S, Aggelis G (2010) Yarrowia lipolytica: a model microorganism used for the production of tailor-made lipids. Eur J Lipid Sci Technol 112:639–654

    Article  CAS  Google Scholar 

  • Rymowicz W, Fatykhova AR, Kamzolova SV, Rywinska A, Morgunov IG (2010) Citric acid production from glycerol-containing waste of biodiesel industry by Yarrowia lipolytica in batch, repeated batch, and cell recycle regimes. Appl Microbiol Biotechnol 87:971–979

    Article  PubMed  CAS  Google Scholar 

  • Rywińska A, Juszczyk P, Wojtatowicz M, Robak M, Lazar Z, Tomaszewska L, Rymowicz W (2013) Glycerol as a promising substrate for Yarrowia lipolytica biotechnological applications. Biomass Bioenergy 48:148–166

    Article  Google Scholar 

  • Sauer M, Porro D, Mattanovich D, Branduardi P (2008) Microbial production of organic acids: expanding the markets. Trends Biotechnol 26(2):100–108

    Article  PubMed  CAS  Google Scholar 

  • Son ED, Choi GH, Kim H, Lee B, Chang IS, Hwang JS (2007) Alpha-ketoglutarate stimulates procollagen production in cultured human dermal fibroblasts, and decreases UVB-induced wrinkle formation following topical application on the dorsal skin of hairless mice. Biol Pharm Bull 30:1395–1399

    Article  PubMed  CAS  Google Scholar 

  • Stottmeister U, Weissbrodt E (1991) Product formation by Yarrowia lipolytica—some generalizing aspects. In: Metabolism alkanow i cwerchsintes produktow mikroorganismami. Sbornik naushnich trudow, Pushino, p. 147–158

  • Stottmeister U, Behrens U, Weissbrodt E, Barth G, Franke-Rinker D, Schulze E (1982) Utilization of paraffins and other noncarbohydrate carbon sources for microbial citric acid synthesis. Z Allg Mikrobiol 22:399–424

    Article  PubMed  CAS  Google Scholar 

  • Stottmeister U, Aurich A, Wilde H, Andersch J, Schmidt S, Sicker D (2005) White biotechnology for Green chemistry: fermentative 2-oxocarboxylic acids as novel building blocks for subsequent chemical syntheses. J Ind Microbiol Biotechnol 32:651–664

    Article  PubMed  CAS  Google Scholar 

  • Weissbrodt E, Barth G, Weber H, Stottmeister U, Duresch R, Richter P (1988) Production of 2-oxoglutaric acid by yeasts. Patent DD 267999

  • Willke T, Vorlop K-D (2004) Industrial bioconversion of renewable resources as an alternative to conventional chemistry. Appl Microbiol Biotechnol 66:131–142

    Article  PubMed  CAS  Google Scholar 

  • Yin X, Madzak C, Du G, Zhou J, Chen J (2012) Enhanced alpha-ketoglutaric acid production in Yarrowia lipolytica WSH-Z06 by regulation of the pyruvate carboxylation pathway. Appl Microbiol Biotechnol 96:1527–1537

    Article  PubMed  CAS  Google Scholar 

  • Yu Z, Du G, Zhou J, Chen J (2012) Enhanced α-ketoglutaric acid production in Yarrowia lipolytica WSH-Z06 by an improved integrated fed-batch strategy. Bioresour Technol 114:597–602

    Article  PubMed  CAS  Google Scholar 

  • Yuzbashev TV, Yuzbasheva EY, Sobolevskaya TI, Laptev IA, Vybornaya TV, Larina AS, Matsui K, Fukui K, Sineoky SP (2010) Production of succinic acid at low pH by a recombinant strain of the aerobic yeast Yarrowia lipolytica. Biotechnol Bioeng 7(4):673–682

    Article  Google Scholar 

  • Zhou J, Zhou H, Du G, Liu L, Chen J (2010) Screening of a thiamine-auxotrophic yeast for α-ketoglutaric acid overproduction. Lett Appl Microbiol 51:264–271

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank Mrs. Maria N. Chiglintseva, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms for her technical assistance in conducting the enzymatic KGA assay. This work was supported by the Ministry of Education and Science of the Russian Federation in the scope of Federal Target Program "Research and development for priority directions of science and technology complex of Russia for 2007–2013" (project no. 14.512.11.0045 from 03.04.2013).

Conflict of interest

The authors have declared no conflict of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Igor G. Morgunov.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Morgunov, I.G., Kamzolova, S.V. & Samoilenko, V.A. Enhanced α-ketoglutaric acid production and recovery in Yarrowia lipolytica yeast by effective pH controlling. Appl Microbiol Biotechnol 97, 8711–8718 (2013). https://doi.org/10.1007/s00253-013-5170-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-013-5170-9

Keywords

Navigation