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Background: Pectinases are a group of enzymes that break down pectin, a polysaccharide that is found in plant cell walls. Today, the application of pectinolytic enzymes plays an important role in food technology for the maceration of fruits and vegetables, including for the extraction and clarification of juice.

Aim of the Study: This study aimed to test the clarifying ability of pectinase of three Yarrowia lipolytica strains (Tias J0-6; YA J3-1; Buy J2-1) identified for cocoa juice in fermentation.

Materials and Methods: The pectinase from Yarrowia lipolytica strains (Tias J0-6; YA J3-1; Buy J2-1) were produced using a basic liquid medium and partially purified through ion exchange chromatography. The fruit juices were clarified by enzymatic treatments against a control (free enzymatic treatment juice) and physicochemical and phytochemical parameters and antioxidant activities of the clarified juices were determined.

Results: The results revealed that from three strains, Tias J0-6 strain produced pectinase which showed the best clarification activity on pineapple and orange juices. The juices clarified exhibited an increasing of clarity values and decreasing viscosity and pH values. Thus, pectinase of Tias J0-6 allowed increasing clarity to 41.81% and 7.27%, to decrease viscosity up to 14.03% and 6.55% respectively for pineapple juices and orange juices clarified. The pH values decreased from 4.036±00 to 4.031±00 and from 5.010±00 to 4.592±00  respectively for orange and pineapple juices. The antioxidant activities of juices clarified were strongly correlated to total tannins and total flavonoids (r = 0.717, r = 0.754, r = 0.855).

Conclusion: Tias J0-6 strain seemed a suitable candidate to biotechnology applications such as fruit juices clarification.

Pectinase, Yarrowia lipolytica, pineapple juice, orange juice, clarification, functional food

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How to Cite
CAMARA, F., MIAN, T. M.-A. S., COULIBALY, W. H., N’GUESSAN, A. R., & BEUGRÉ, G. A. M. (2022). CLARIFICATION TREATMENTS OF PINEAPPLE (Malus domestica) AND ORANGE (Citrus sinensis) JUICES BY PECTINASE FROM Yarrowia lipolytica STRAINS IDENTIFIED FROM COCOA JUICE IN FERMENTATION. Journal of Biochemistry International, 9(4), 82-93.
Original Research Article


Fogarty MV. and Kelly CT. In Microbial Enzymes and Biotechnology. ed. Fogarty, M.W. pp. 131±182. London & New York: Elsevier Applied Science Publishers. ISBN 1-85166486-6 Gainvors, A. and Belarbi, A. 1995 Detection methods for polygalacturonase producing strains of Saccharomyces cerevisae. Yeast. 1983;10: 1311-1319.

Mohnen D. Pectin structure and biosynthesis. Curr Opin Plant Biol. 2008; 11:266–277.

Yadav PK, Singh VK, Yadav S, Yadav KDS, Yadav D. In silico analysis of pectin lyase and pectinase sequences. Biochem (Moscow). 2009;74(9):1049–1055.

Reddy PL, Sreeramulu A. Isolation, identification and screening of pectinolytic fungi from different soil samples of Chittoor district. Int J of Life Sciences Biotechnology and Pharma Research. 2012;1(3):186-193.

Yadav D, Yadav S, Anand G. Production, purification and biochemical characterization of an exo-polygalacturonase from Aspergillus niger MTCC 478 suitable for clarification of orange juice. Biotech, 2017;7: 3-8.

Okonji RE, Itakorode BO, Ovumedia JO, Adedeji OS. Purification and biochemical characterization of pectinase produced by Aspergillus fumigatus isolated from soil of decomposing plant materials. J App Biol Biotech. 2019;7: 1-8.

Aaisha GA, Barate DL. Isolation and identification of pectinolytic bacteria from soil samples of Akola Region, India. Int J of Current Microbiology and Applied Sciences. 2015;5(1): 514-521.

Meryandini A, Cocok AM. Fahrurrozi B. Pectinase production and clarification treatments of apple (malus domestica) juice. Annales Bogorienses. 2017;21: 63-68.

Bouatenin KMJP, Djeni NT, Kouame KA, Coulibaly WH, Dje KM. Excretion of β-glucosidase and pectinase by microorganisms isolated from cassava traditional ferments used for attieke production in Côte d'Ivoire. Biocatalysis and Agricultural Biotechnology. 2019;20:101217.

Djoulde RD. Etoa FX, Essia Ngang JJ, Mbofung CMF. Screening des microorganismes à potentialités fermentaires pour le manioc. Tropicultura. 2005;23:11-18.

Merin MG, Martin MC, Rantsior K, Cocolin L, Morata de Ambrosini VI. Characterization of pectinase activity for enology from yeasts occurring in Argentine bonarda grape. Braz J Microbiol. 2015;46:815-823.

Dzogbefia VP, Ametefe GD, Kwatia CAS. Optimal conditions for pectinase production by Saccharomyces cerevisiae (ATCC 52712) in solid state fermentation and its efficacy in orange juice extraction. Journal of Biotechnology and Biochemistry. 2017;3:78-86.

Poondla V, Bandikari R, Subramanyam R, Obulam VSR. Low temperature active pectinases production by Saccharomyces cerevisiae isolates and their characterization, Biocatalysis and Agricultural Biotechnology. 2015;4:70-76.

Castilho LR, Medronho RA, Alves TLM. Production and extraction of pectinases obtained by solid state fermentation of agroindustrial residues with Aspergillus niger. Bioresour Technol. 2000 ;71:45–50.

Silva D, Martins ES, Silva R, Gomes E. Pectinase production by Penicillium viridicatum RFC3 by solid state fermentation using agricultural wastes and agro-industrial byproducts. Br J Microbiol. 2002;33:318–324.

Pedrolli DB, Carmona EC. Pectin lyase from Aspergillus giganteus: comparative study of productivity of submerged fermentation on citrus pectin and orange waste. Prikl Biokhim Mikrobiol. 2009;45:677–683.

Dinu D, Nechifor MT, Stoian G, Costache M, Dinischiotu A. Enzymes with new biochemical properties in the pectinolytic complex produced by Aspergillus niger MIUG 16. J Biotechnol. 2007;131:128–137

Singh C. Poonia GS. Toor GS. Distribution pattern of fruit processing industry in Punjab. Indian Food Packer. 1994; 8(1): 47.

Vaillant F, Millan P, Brien GO, Dornier M, Decloux M, Reynes M. Crossow microfiltration of passion fruit juice after partial enzymatic liquefaction. Journal of Food Engineering. 1999 ;42:215-224.

Tribess TB, Tadini CC. Inactivation kinetics of pectin methyl esterase in orange juice as a function of pH and temperature-time process conditions. Journal of the Science of Food and Agriculture. 2006;86:1328-1335.

Sulaiman MZ, Sulaiman NM, Liew SY. Limiting permeate flux in the clarification of untreated starfruit juice by membrane ultrafiltration. Chemical Engineering Journal. 1998; 69:145-148.

Kashyap DR, Vohra PK, Chopra S, Tewari R. Applications of Pectinases in commercial sector: a review. Biores Technol. 2001;77:215–227.

Landbo A-K, Meyer AS. Statistically designed two step response surface optimizations of enzymatic prepress treatment to increase juice yield and lower turbidity of elderberry juice. Innovative Food Science and Emerging Technologies. 2007; 8:135-142.

Blanco P, Sierro C, Villa TG. Production of pectic enzymes in yeasts. FEMS Microbiology Letters. 1999;175:1-9.

Mutlu M, Sarioglu K, Demir N, Ercan MT, Acar J. The use of commercial pectinase in fruit juice industry. Part I: viscosimetric determination of enzyme activity. Journal of Food Engineering. 1999;41:147-150.

Kaur G, Kumar S, Satyanarayana T. Production, characterization and application of a thermostable polygalacturonase of a thermophilic moul Sporotrichum Termophile. Bioresource Technology. 2004;94:239-243.

Abdullah LAG, Sulaiman NM, Aroua MK, Noor, MMMJ. Response surface optimization of conditions for clarification of carambola fruit juice using a commercial enzyme. Journal of Food Engineering. 2007;81:65-71.

Mian TM-AS, Camara F, Coulibaly WH, Beugré GAM, N’Guessan AR, Djè KM. Identification of pectinase-producing yeast strains isolated from fermented mucilage of cocoa juice. Current topics in biotechnology. 2021;12:27–38.

Miller GL, Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem. 1959; 31(3):426–428.

Acar J, Alper N, Esturk O. The production of cloudy apple nectar using total liquefaction enzymes. Fruit Process. 1999;8:314-317.

Singleton V, Orthofer R, Lamuela-Ravento´s R. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. In L. Packer (Ed.). Oxidants and antioxidants, part A, methods in enzymology (Vol. 299, pp. 152–178). 1999. New York: Academic Press

Ribereau-Gayon P, Stonestreet E. Dosage des tanins du vin rouge et détermination de leur structure. Chim. Anal. 1966; 48: 188–196.

Meda A, Lamien CE, Romito M, Millogo J, Nacoulma OG. Determination of total phenolic, flavonoid and proline contents in Burkina Faso honeys as well as their radical scavenging activity. Food Chemistry. 2005; 91: 571-577.

Ribereau-Gayon P, Stonestreet E. Le dosage des anthocyanes dans le vin rouge. Bull. Soc. Chim. 1965 ; 9 : 2649–2652.

Alves CQ, David JM, David, JP, Bahia MV, Aguiar RM. Métodos para determinação de atividade antioxidante in vitro em substratos orgânicos. Química Nova. 2010; 33: 2202–2210.

Brand-Williams W, Cuvelier ME, Berset C. Use of free radical method to evaluate antioxidant activity. Lebensm. Wiss.Technol. 1995; 28: 25-30.

Lim L, Sun Y, Laura T, Liang X, Ye H, Zeng X. Determination of polyphenolic content and antioxidant activity of Kudingcha made from Ilex kudingcha C.J. Tseng. Food Chem. 2009; 112: 35–41.

Utami R, Widowati E, Christy A. Screening and characterization of amylase enzyme in sweet orange (Citrus sinensis) juice clarification. Nusantara bioscience. 2016;8 (2):268-272.

Garg G, Singh A, Kaur A, Singh R, Kaur J, Mahajan R. Microbial pectinases : an ecofriendly tool of nature for industries. 3 Biotech. 2016;6(1) :47.

Rai P, Rai C, Majumdar GC, Dasgupta S, De S. Resistance in series model for ultrafiltration of mosambi (Citrus sinensis (L.) Osbeck) juice in a stirred continuous mode. Journal of Membrane Science. 2006 ;283:116-122.

Dedehou SCAE., Dossou J, Soumanou MM. Etude diagnostique des technologies de transformation de la pomme de cajou en jus au Bénin. International Journal of Biological and Chemical Sciences. 2015;9(1):371-387.

Sharma HP, Sharma S, Vaishali Prasad K. Application of Non-thermal clarification in Fruit Juice processing - A Review. South Asian Journal of Food Technology and Environment. 2015;1(1): 15- 21

Nakkeeran E, Umesh-Kumar S, Subramanian R. Aspergillus carbonarius polygalacturonases purified by integrated membrane process and affinity precipitation for apple juice production. Biores Technol. 2011;102:3293–3297

Tu T, Luo H, Meng K, Cheng Y, Ma R, Shi P, Huang H, Bai Y, Wang Y, Zhang L, Yao B. Improvement in thermostability of an Achaetomium sp. Strain Xz8 endopolygalacturonase via the optimization of charge–charge interactions. Appl Environ Micrbiol. 2015; 81(19): 6938–6944.

Barman S, Sit N, Badwaik LS, Deka SC. Pectinase production by Aspergillus niger using banana (Musa balbisiana) peel as substrate and its effect on clarification of banana juice. J Food Sci Technol. 2015;52:3579–3589.

Pan X, Li K, Ma R, Shi P, Huang H, Yang P, Meng K, Yao B. Biochemical characterization of three distinct polygalacturonases from Neosartorya fischeri P1. Food Chem. 2015;188:569–575.

Dey TB, Adak S, Bhattacharya P, Banerjee R. Purification of polygalacturonase from Aspergillus awamori Nakazawa MTCC 6652 and Its application in apple juice clarification. Food Sci Technol. 2014;59:591–595.

Arsad P, Sukor R, Wan IWZ, Mustapha NA, Meor AS. Effect of enzymatic treatment on physicochemical properties of sugar palm fruit juice. Int J on Adv Science Engineering Information Technology. 2015;5:308-312.

Alvarez S, Alvarez R, Riera FA, Coca J. Influence of depectinization on apple juice ultra-filtration colloidal and surface. Physicochemical and Engineer Aspect. 1998;138:337-382.

Wojdyło A, Oszmianski J, Kolniak J. Effect of pectinase treatment on extraction of antioxidant phenols from pomace, for the production of puree-enriched cloudy apple juices. Food Chemistry. 2011;127:623–631.

Iriani ES, Said EG, Suryani AS. Pengaruh Konsentrasi Penambahan Pektinase dan Kondisi Inkubasi terhadap Rendemen dan Mutu Jus Mangga Kuini (mangifera odorata Griff). J Pascapanen. 2005;2(1):11-17.

Poletto P, Borsoi C, Zeni M, da Silveira MM. Downstream processing of pectinase produced by Aspergillus niger in solid state cultivation and its application to fruit juices clarification. Food Sci Technol. 2015;35(2):391-397.

Müller L, Gnoyke S, Popken AM, Böhma V. Antioxidant capacity and related parameters of different fruit formulations. LWT- Food Science and Technology. 2010;43:992-999.

Halliwell B, Gutteridge JMC. In Free Radicals in Biology and Medicine (3rd ed.). New York: 1999. Oxford University Press.

Siebert JK. Haze formation in beverages. LWT-Food Science and Technology. 2006; 39:987-994.

Pinelo M, Zeuner B, Meyer AS. Juice clarification by protease and pectinase treatments indicates new roles of pectin and protein in cherry juice turbidity. Food and Bioproducts Processing. 2010;88:259-265.

Wojdyło A, Oszmianski J, Kolniak J. Effect of pectinase treatment on extraction of antioxidant phenols from pomace, for the production of puree-enriched cloudy apple juices. Food Chemistry. 2011;127:623–631.