Enhancing erythritol productivity in Yarrowia lipolytica using metabolic engineering

[en] Erythritol (1,2,3,4-butanetetrol) is a four-carbon sugar alcohol with sweetening properties that is used by the agrofood industry as a food additive. In this study, we demonstrated that metabolic engineering can be used to improve the production of erythritol from glycerol in the yeast Yarrowia lipolytica. The best results were obtained using a mutant that overexpressed GUT1 and TKL1, which encode a glycerol kinase and a transketolase, respectively, and in which EYK1, which encodes erythrulose kinase, was disrupted; the latter enzyme is involved in an early step of erythritol catabolism. In this strain, erythritol productivity was 75% higher than in the wild type; furthermore, the culturing time needed to achieve maximum concentration was reduced by 40%. An additional advantage is that the strain was unable to consume the erythritol it had created, further increasing the process's efficiency. The erythritol productivity values we obtained here are among the highest reported thus far.

Disciplines :

Biotechnology

Author, co-author :

Carly, Frédéric

Vandermies, Marie ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbial, food and biobased technologies

Telek, Samuel ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbial, food and biobased technologies

Steels, Sébastien ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbial, food and biobased technologies

Thomas, Stéphane

Nicaud, Jean-Marc

Fickers, Patrick ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbial, food and biobased technologies

Language :

English

Title :

Enhancing erythritol productivity in Yarrowia lipolytica using metabolic engineering

Publication date :

July 2017

Journal title :

Metabolic Engineering

ISSN :

1096-7176

eISSN :

1096-7184

Publisher :

Academic Press

Volume :

Pages :

19-24

Peer reviewed :

Peer Reviewed verified by ORBi

Funders :

FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture [BE]

Available on ORBi :

since 18 July 2017

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Bibliography

Almeida, J.R.M., Fávaro, L.C.L., Quirino, B.F., Biodiesel biorefinery: opportunities and challenges for microbial production of fuels and chemicals from glycerol waste. Biotechnol. Biofuels, 5, 2012, 48.
Bernt, W.O., Borzelleca, J.F., Flamm, G., Munro, I.C., Erythritol: a review of biological and toxicological studies. Regul. Toxicol. Pharmacol. 24 (1996), S191–S197.
Barth, G., Gaillardin, C., Yarrowia lipolytica. In Nonconventional Yeasts in Biotechnology. 1996, Springer Berlin Heidelberg, Berlin, Heidelberg, 313–388.
Carly, F., Gamboa-Melendez, H., Vandermies, M., Damblon, C., Nicaud, J.-M., Fickers, P., Identification and characterization of EYK1, a key gene for erythritol catabolism in Yarrowia lipolytica. Appl. Microbiol. Biotechnol., 2017 (submitted for publication).
Fickers, P., Benetti, P., Wache, Y., Marty, A., Mauersberger, S., Smit, M., Nicaud, J.-M., Hydrophobic substrate utilisation by the yeast Yarrowia lipolytica, and its potential applications. FEMS Yeast Res. 5 (2005), 527–543.
Ghezelbash, G.R., Nahvi, I., Malekpour, A., Erythritol production with minimum by-product using Candida magnoliae mutant. Appl. Biochem. Microbiol. 50 (2014), 292–296.
Hallsworth, J.E., Magan, N., A rapid HPLC protocol for detection of polyols and trehalose. J. Microbiol. Methods 29 (1997), 7–13.
Higuchi, R., Krummel, B., Saiki, R.K., A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 16 (1988), 7351–7367.
Ishizuka, H., Wako, K., Kasumi, T., Sasaki, T., Breeding of a mutant of Aureobasidium sp. with high erythritol production. J. Ferment. Bioeng. 68 (1989), 310–314.
Jeya, M., Lee, K.-M., Tiwari, M.K., Kim, J.-S., Gunasekaran, P., Kim, S.-Y., Kim, I.-W., Lee, J.-K., Isolation of a novel high erythritol-producing Pseudozyma tsukubaensis and scale-up of erythritol fermentation to industrial level. Appl. Microbiol. Biotechnol. 83 (2009), 225–231.
Kawanabe, J., Hirasawa, M., Takeuchi, T., Oda, T., Ikeda, T., Noncariogenicity of erythritol as a substrate. Caries Res. 26 (1992), 358–362.
Kim, S.-Y., Lee, K.-H., Kim, J.-H., Oh, D.-K., Erythritol production by controlling osmotic pressure in Trigonopsis variabilis. Biotechnol. Lett. 19 (1997), 727–729.
Kuznetsova, E., Proudfoot, M., Gonzalez, C.F., Brown, G., Omelchenko, M.V., Borozan, I., Carmel, L., Wolf, Y.I., Mori, H., Savchenko, A.V., et al. Genome-wide analysis of substrate specificities of the Escherichia coli haloacid dehalogenase-like phosphatase family. J. Biol. Chem. 281 (2006), 36149–36161.
Lee, J.-K., Ha, S.-J., Kim, S.-Y., Oh, D.-K., Increased erythritol production in Torula sp. by Mn2+ and Cu2+. Biotechnol. Lett. 22 (2000), 983–986.
Lee, D.-H., Lee, Y.-J., Ryu, Y.-W., Seo, J.-H., Molecular cloning and biochemical characterization of a novel erythrose reductase from Candida magnoliae JH110. Microb. Cell Factor., 9, 2010, 43.
Lin, S.-J., Wen, C.-Y., Wang, P.-M., Huang, J.-C., Wei, C.-L., Chang, J.-W., Chu, W.-S., High-level production of erythritol by mutants of osmophilic Moniliella sp. Process Biochem. 45 (2010), 973–979.
Livak, K.J., Schmittgen, T.D., Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25 (2001), 402–408.
Mirończuk, A.M., Furgała, J., Rakicka, M., Rymowicz, W., Enhanced production of erythritol by Yarrowia lipolytica on glycerol in repeated batch cultures. J. Ind. Microbiol. Biotechnol. 41 (2014), 57–64.
Mirończuk, A.M., Rakicka, M., Biegalska, A., Rymowicz, W., Dobrowolski, A., A two-stage fermentation process of erythritol production by yeast Y. lipolytica from molasses and glycerol. Bioresour. Technol. 198 (2015), 445–455.
Mirończuk, A.M., Rzechonek, D.A., Biegalska, A., Rakicka, M., Dobrowolski, A., A novel strain of Yarrowia lipolytica as a platform for value-added product synthesis from glycerol. Biotechnol. Biofuels 9 (2016), 180–191.
Moon, H.-J., Jeya, M., Kim, I.-W., Lee, J.-K., Biotechnological production of erythritol and its applications. Appl. Microbiol. Biotechnol. 86 (2010), 1017–1025.
Munro, I., Bernt, W., Borzelleca, J., Flamm, G., Lynch, B., Kennepohl, E., Bär, E., Modderman, J., Erythritol: an interpretive summary of biochemical, metabolic, toxicological and clinical data. Food Chem. Toxicol. 36 (1998), 1139–1174.
Nicaud, J.-M., Yarrowia lipolytica. Yeast 29 (2012), 409–418.
Paradowska, K., Nitka, D., Purification and characterization of erythritol dehydrogenase from Mycobacterium smegmatis. Ann. UMCS Pharm. 22 (2009), 47–55.
Park, Y.-C., Oh, E.J., Jo, J.-H., Jin, Y.-S., Seo, J.-H., Recent advances in biological production of sugar alcohols. Curr. Opin. Biotechnol. 37 (2016), 105–113.
Querol, A., Barrio, E., Huerta, T., Ramón, D., Molecular monitoring of wine fermentations conducted by active dry yeast strains. Appl. Environ. Microbiol. 58 (1992), 2948–2953.
Rakicka, M., Rywińska, A., Cybulski, K., Rymowicz, W., Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants. Braz. J. Microbiol. 47 (2016), 417–423.
Rymowicz, W., Rywińska, A., Gładkowski, W., Simultaneous production of citric acid and erythritol from crude glycerol by Yarrowia lipolytica Wratislavia K1. Chem. Pap. 62 (2008), 239–246.
Ryu, Y.-W., Park, C.Y., Park, J.B., Kim, S.-Y., Seo, J.-H., Optimization of erythritol production by Candida magnoliae in fed-batch culture. J. Ind. Microbiol. Biotechnol. 25 (2000), 100–103.
Rywińska, A., Juszczyk, P., Wojtatowicz, M., Robak, M., Lazar, Z., Tomaszewska, L., Rymowicz, W., Glycerol as a promising substrate for Yarrowia lipolytica biotechnological applications. Biomass-Bioenergy 48 (2013), 148–166.
Sambrook, J., Fritsch, E., Maniatis, T., 1989. Molecular Cloning: A Laboratory Manual: Vol. 2 (S.l.: Cold Spring Harbor).
Sassi, H., Delvigne, F., Kar, T., Nicaud, J.-M., Coq, A.-M.C.-L., Steels, S., Fickers, P., Deciphering how LIP2 and POX2 promoters can optimally regulate recombinant protein production in the yeast Yarrowia lipolytica. Microb. Cell Factor. 15 (2016), 159–169.
Sawada, K., Taki, A., Yamakawa, T., Seki, M., Key role for transketolase activity in erythritol production by Trichosporonoides megachiliensis SN-G42. J. Biosci. Bioeng. 108 (2009), 385–390.
Tomaszewska, L., Rywińska, A., Gładkowski, W., Production of erythritol and mannitol by Yarrowia lipolytica yeast in media containing glycerol. J. Ind. Microbiol. Biotechnol. 39 (2012), 1333–1343.
Vandermies, M., Denies, O., Nicaud, J.M., Fickers, P., EYK encoding erythrulose kinase as a catabolic selectable marker for genome editing in the non-conventional yeast Yarrowia lipolytica. J. Microb. Methods, 2017 (in press).
Yang, L.-B., Dai, X.-M., Zheng, Z.-Y., Zhu, L., Zhan, X.-B., Lin, C.-C., Proteomic analysis of erythritol-producing Yarrowia lipolytica from glycerol in response to osmotic pressure. J. Microbiol. Biotechnol. 7 (2015), 1059–1069.
Yang, L.-B., Zhan, X.-B., Zheng, Z.-Y., Wu, J.-R., Gao, M.-J., Lin, C.-C., A novel osmotic pressure control fed-batch fermentation strategy for improvement of erythritol production by Yarrowia lipolytica from glycerol. Bioresour. Technol. 151 (2014), 120–127.
Zinjarde, J., Food-related application of Yarrowia lipolytica. Food Chem. 152 (2014), 1–10.