THE OPTIMUM CONDITIONS FOR FRUCTOOLIGOSACCHARIDE PRODUCTION BY USING CRUDE FRUCTOSYLTRANSFERASE OF PLANT ORIGIN

Main Article Content

AHMED ABDULKAREEM IBRAHEM
WASAN KADHIM A. AL-TEMIMI
SARMAD GHAZI AL-SHAWI

Abstract

Fructooligosaccharide (FOS) has an economic importance for its multiple uses in the food and pharmaceutical industries. FOS is produced naturally or industrially by enzymatic catalysis of a plant or microbial Fructosyltransferase (Ftase) enzyme. In the current study, the enzyme was extracted from plant sources, including Iraqi radish (Raphanus sativus var. Longipinnatus), Iraqi garlic (Allium sativum), Iraqi artichoke (Helianthus tuberosus), sweet potato (Ipomoea batatas) and Egyptian pineapple (Ananas comosus). The enzyme was extracted using potassium phosphate buffer at a pH range (6-8) in the presence of Cysteine, Triton x-100, and sodium acetate buffer at a pH range (4-7) in the presence of PMSF, EDTA and mercaptoethanol. Plant extracts showed the highest specific efficacy when using sodium acetate buffer at pH 6 for pineapple residues which was 200.732 units/mg protein. When studying the optimal conditions for FOS production using the crude enzymatic extract of the pineapple residues, which included (pH, temperature, concentration of the substrate, the concentration of the enzymatic extract and time), it was found that the optimal conditions for FOS production depending on the enzymatic activity were (6.5, 40°C, 40% and 250 µL and 18 hours respectively) as the enzymatic activity reached (271.32, 265.385, 266.615, 273.564 and 271.897) units/ml, respectively. By determining the enzymatic reaction products with Thin Layer Chromatography (TLC) and comparing the reaction products with standard sugars, it was found that the extract contains glucose, sucrose, 1-kestose, Nystose and FOS at Rf of (0.83, 0.65, 0.566, 0.48 and 0.41). Respectively, the presence of fructose was not observed.

Keywords:
Fructooligosaccharide, fructosyltransferase, optimal condition, thin layer chromatography

Article Details

How to Cite
IBRAHEM, A. A., AL-TEMIMI, W. K. A., & AL-SHAWI, S. G. (2021). THE OPTIMUM CONDITIONS FOR FRUCTOOLIGOSACCHARIDE PRODUCTION BY USING CRUDE FRUCTOSYLTRANSFERASE OF PLANT ORIGIN. PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY, 22(37-38), 252-265. Retrieved from https://www.ikprress.org/index.php/PCBMB/article/view/6562
Section
Original Research Article

References

Sanchez O, Guio F, Garcia D, Silva E, Caicedo L. Fructooligosaccharides production by Aspergillus sp. N74 in a mechanically agitated airlift reactor. Food and Bioproducts Processing. 2008;86(2):109-115.
DOI: 10.1016/j.fbp.2008.02.003

Sabater-Molina M, Larque E, Torrella F, Zamora S. Dietary fructo oligo saccharides and potential benefits on health. J Physiol Biochem. 2009;65:315–28.

Ramli MAB. Purification of Fructosyl transferase (FTase) From Aspergillus niger to Enhance Production of Fructo oligo saccharides (FOS) as a Food Additive. Thesis submitted to Faculty of Chemical and Natural Resources Engineering, Univ. of Malaysia Pahang; 2012.

Ngampanya B, Keayarsa S, Jaturapiree P, Prakobpran P, Wichienchot S. Characterization of transfructosylating activity enzyme from tubers of tropical Jerusalem artichoke (Helianthus tuberosus L.) for production of fructooligosaccharides. IFRJ. 2016;23(5): 1965-1972.

Antosova M, Polakovic M. Fructosyltransferases: The enzymes catalyzing production of fructooligosaccharides. Areview, Chem. Pap. 2001;55(6):350-358.
Available:https://www.researchgate.net/publication/285819871

Koops AJ, Jonker HH. Purification and Characterization of the Enzymes of Fructan Biosynthesis in Tubers of Helianthus tuberosus Colombia (II. Purification of Sucrose: Sucrose 1-Fructosyltransferase and Reconstitution of Fructan Synthesis in Vitro with Purified Sucrose: Sucrose 1-Fructosyltransferase and Fructan: Fructan 1-Fructosyltransferase). Plant Physiol. 1996;110(4):1167-1175.
DOI: 10.1104/pp.110.4.1167.

Ghazi I, Fernandez-Arrojo L, De Segura AG, Alcalde M. Beet sugar syrup and molasses as low-cost feedstock for the enzymatic production of fructo-oligosaccharides. Journal of Agricultural and Food Chemistry. 2006;54(8):2964-8 .
DOI: 10.1021/jf053023b

Wichienchot S, Prakobpran P, Ngampanya B, Jaturapiree P. Production, purification and fecal fermentation of fructooligosaccharide by FTase from Jerusalem artichoke. International Food Research Journal. 2017;24(1):134-141.

Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry. 1959;31(3):426-428.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry. 1976;72(1-2):248-254.

Sangeetha PT, Ramesh MN, Prapulla SG. Production of fructosyltransferase by Aspergillus oryzae CFR 202 in solid-state fermentation using agricultural by-products. Appl Microbiol Biotechnol. 2004;65:530–537.
DOI: 10.1007/s00253-004-1618-2

Mussatto SI, Aguilar CN, Rodrigues LR, Teixeira JA. Colonization of Aspergillus japonicus on synthetic materials and application to the production of fructooligosaccharides. Carbohydr Res. 2009;344:795–800.

Fara, SOULY. Separation and analysis of some sugars by using thin layer chromatography. JASSBT. 1979;20:251-254.

SPSS. Statistical Package for the Social Sciences. Version, USA. 2019;26.

Segel IH. Biochemical calculations. 2nd Edn, John and sons. Inc.New York; 1976.

Balasundaram B, Pandit AB. Significance of location of enzymes on their release during microbial cells disruption. Biotech. Bioeng. 2001;75:607-614.

NurDini MN, Noormazlinah A, Hamidi NH, Aini MN, Sakinah AM, Ali MF. Enzymaic Production of Fructooligosaccharides from Phytoenzymes of Ananas Comous Waste by using Response Surfacemthodology: On. Journal of Chemical Engineering and Industrial Biotechnology. 2018;3(1):26-36.

Ngampanya B, Keayarsa S, Ngernmeesri K, Prakobtran P, Wichienchot S. Production of fructo-oligosaccharides by partial purified fructosyltransferase from variety of Jerusalem artichoke grown in Thailand. Proceedings of the 7th International Symposium of the Protein Society of Thailand. 2012;172-177.

Ende WVD, Laere AV. Purification and properties of an invertase with sucrose: sucrose fructosyltransferase (SST) activity from the roots of Cichorium intybus L. New Phytol. 1993;123:31-37.

Judprasong K, Tanjor S, Puwastien P, Sungpuag P. Investigation of Thai plants for potential sources of inulin-type fructans. J. Food Composition and Analysis. 2011;24:624-649.

Tanjor S, Judprasong K, Chaito C, Jogloy S. Inulin and fructooligosacharides in different varieties of Jerusalem artichoke (Helianthus tuberosus L.). KKU Res. J. 2012;17(11):25-34.

Fernández RC, Maresma BG, Juárez A, Martínez J. Production of fructooligosaccharides by β‐fructofuranosidase from Aspergillus sp 27H. Journal of Chemical Technology & Biotechnology: International Research in Process, Environmental & Clean Technology. 2004;79(3):268-272.

Jung KH, Kim JH, Jeon YJ, Lee JH. Production of high fructo-oligosaccharide syrup with two enzyme system of fructosyltransferase and glucose oxidase. Biotechnol. Lett. 1993;15:65-70.

Yun JW, Lee MG, Song SK. Batch production of high-content fructo-oligosaccharides from sucrose by the mixed-enzyme system of β-fructofuranosidase and glucose oxidase. J. Fermentation and Bioengineering. 1994;77(2):159-163.

Deepa C Khandekar, Tapas Palai Aman Agarwal, Prashant K Bhattacharya. Kinetics of sucrose conversion to fructo-oligosaccharides using enzyme (invertase) under free condition. Bioprocess and Biosystem Engineering. 2014;37:2529-2537.

Dhake AB, Patil MB. Effect of substrate feeding on production of fructosyltransferase by Penicillium purpurogenum. Brazilian Journal of Microbiology. 2007;38(2):194-199.

Kupcova E, Reiffová K, Bazeľ Y. Thin-layer chromatography: An efficient method for the optimization of dispersive liquid-liquid microextraction. Acta Chimica Slovenica. 2018;65(2):388-393.

Fontana JD, Grzybowski A, Tiboni M, Passos M. Fructo-oligosaccharide production from inulin through partial citric or phosphoric acid hydrolyses. Journal of Medicinal Food. 2011;14(11):1425-1430.

Lu L, Wu J, Song D, Zhao H, Gu G, Guo Y, et al. Purification of fructooligosaccharides by immobilized yeast cells and identification of ethyl β-d-fructofuranoside as a novel glycoside formed during the process. Bioresource Technology. 2013;132:365-369.

Chikkerur J, Ashis Kumar Samanta, Kolte AK AP, Dhali A, Roy S. Production of Short Chain Fructo-oligosaccharides from Inulin of Chicory Root Using Fungal Endoinulinase. Applied Biochemistry and Biotechnology. 2019; 191:695-715.
DOI: org/10.1007/s/2010-019-03215-7