Kinetic Study of Erythrose Reductase Extracted from Yarrowia lipolytica

Journal of Cell and Molecular Research

مقاله 6، دوره 10، شماره 2 - شماره پیاپی 20، خرداد 2019، صفحه 97-101   

نوع مقاله: Research Articles

شناسه دیجیتال (DOI): 10.22067/jcmr.v10i2.73964

نویسندگان

Masoud Mohammadi Farsani؛ Mohammad Mohammadi^* ؛ Gholam Reza Ghezelbash؛ Ali Shahriari

Shahid Chamran University of Ahvaz, Ahvaz, Iran

چکیده

     Erythritol as a non-caloric and non-cariogenic sweetener is safe for diabetics. Both microbial fermentation and chemical methods can be used to produce erythritol, but chemical methods failed to be industrialized due to their low efficiency. Moniliella tomentosa, Aureobasidium sp. and Yarrowia lipolytica are industrial producers of erythritol. Erythrose reductase (ER) is a key enzyme in the biosynthesis of erythritol and catalyzes the final step in this pathway. Enzyme extract was obtained from Y. lipolytica by grinding cells with 0.5mm glass beads and ER activity was performed using 10 µl enzyme extract, 7.5 mM NADPH and 12 mM D-erythrose in potassium phosphate buffer (pH 7.5). Reaction was monitored with decreasing of NADPH absorbance in OD340 at 37 ˚C for 8 min by a microplate analyzer. In order to determine the activation energy (Ea), activity of enzyme was measured in 4-45 ˚C and results were analyzed with Kinetic software according to Arrhenius equation. The best enzyme activity of ER was 6.268 mU. One unit of ER activity was defined as the amount of enzyme that catalyzes the oxidation of 1μmol of NADPH per minute. Specific activity of enzyme was equal to 3.24U/mg and finally the Ea was determined to be 29.6208 KJ.ER specific activity in this study was lower than the only similar study that used Y. lipolytica. Purification, overexpression and optimizing the reaction can help to increase enzyme performance.

کلیدواژه‌ها

erythrose reductase؛ Yarrowia lipolytica؛ enzyme kinetics

مراجع

Dobrowolski A., Mituła P., Rymowicz W. and Mirończuk A. M. (2016) Efficient conversion of crude glycerol from various industrial wastes into single cell oil by yeast Yarrowia lipolytica. Bioresource technology 207:237-243.

Ghezelbash G., Nahvi I. and Rabbani M. (2012) Study of polyols production by Yarrowia lipolytica in batch culture and optimization of growth condition for maximum production. Jundishapur Journal of Microbiology 5:546-549.

Ghezelbash G. R., Nahvi I. and Emamzadeh R. (2014) Improvement of erythrose reductase activity, deletion of by-products and statistical media optimization for enhanced erythritol production from Yarrowia lipolytica mutant 49. Current microbiology 69:149-157.

Groenewald M., Boekhout T., Neuveglise C., Gaillardin C., Van Dijck P. W. and Wyss M. (2014) Yarrowia lipolytica: safety assessment of an oleaginous yeast with a great industrial potential. Critical reviews in microbiology 40:187-206.

Ishizuka H., Tokuoka K., Sasaki T. and Taniguchi H. (1992) Purification and some properties of an erythrose reductase from an Aureobasidium sp. mutant. Bioscience, biotechnology, and biochemistry 56:941-945.

Ishizuka H., Wako K., Kasumi T. and Sasaki T. (1989) Breeding of a mutant of Aureobasidium sp. with high erythritol production. Journal of Fermentation and Bioengineering 68:310-314.

Janek T., Dobrowolski A., Biegalska A. and Mirończuk A. M. (2017) Characterization of erythrose reductase from Yarrowia lipolytica and its influence on erythritol synthesis. Microbial Cell Factories 16:118.

Jovanović B., Mach R. L. and Mach-Aigner A. R. (2013) Characterization of erythrose reductases from filamentous fungi. AMB Express 3:43.

Kamzolova S. V., Vinokurova N. G., Lunina J. N., Zelenkova N. F. and Morgunov I. G. (2015) Production of technical-grade sodium citrate from glycerol-containing biodiesel waste by Yarrowia lipolytica. Bioresource technology 193:250-255.

Lee D. H., Lee Y. J., Ryu Y. W. and Seo J. H. (2010) Molecular cloning and biochemical characterization of a novel erythrose reductase from Candida magnoliae JH110. Microbial cell factories 9:43.

Lee J. K., Kim S. Y., Ryu Y. W., Seo J.H. and Kim J.H. (2003a) Purification and characterization of a novel erythrose reductase from Candida magnoliae. Applied and environmental microbiology 69:3710-3718.

Lee J. K., Hong K. W. and Kim S. Y. (2003b) Purification and properties of a NADPHdependent erythrosereductase from the Newly isolated Torula corallina. Biotechnology progress 19:495-500.

Moon H.J., Jeya M., Kim I.W. and Lee J.K. (2010) Biotechnological production of erythritol and its applications. Applied microbiology and biotechnology 86:1017-1025.

Morgunov I. G., Kamzolova S. V. and Lunina J. N. (2013) The citric acid production from raw glycerol by Yarrowia lipolytica yeast and its regulation. Applied microbiology and biotechnology 97:7387-7397.

Park E. H., Ryu Y. W. and Kim M. D. (2011) Role of osmotic and salt stress in the expression of erythrose reductase in Candida magnoliae. Journal of microbiology and biotechnology 21:1064-1068.

Rymowicz W., Rywińska A. and Marcinkiewicz M. (2009) High-yield production of erythritol from raw glycerol in fed-batch cultures of Yarrowia lipolytica. Biotechnology Letters 31:377-380.

Tomaszewska L., Rakicka M., Rymowicz W. and Rywińska A. (2014a) A comparative study on glycerol metabolism to erythritol and citric acid in Yarrowia lipolytica yeast cells. FEMS yeast research 14:966-976.

Tomaszewska L., Rymowicz W. and Rywińska A. (2014b) Mineral supplementation increases erythrose reductase activity in erythritol biosynthesis from glycerol by Yarrowia lipolytica. Applied biochemistry and biotechnology 172:3069-3078.

Tomaszewska L., Rywińska A. and Gładkowski W. (2012) Production of erythritol and mannitol by Yarrowia lipolytica yeast in media containing glycerol. Journal of industrial microbiology & biotechnology 39:1333-1343.