Partial Purification and Kinetic Properties of Polygalacturonase from Chrysophyllum albidum G. Don Fruit doi.org/10.26538/tjnpr/v5i11.18
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Abstract
Polygalacturonase is the major pectic enzyme responsible for hydrolysing pectic substances into their monomeric units. This study evaluated the kinetic properties of extracted and partially purified polygalacturonase from Chrysophyllum albidum fruit. Polygalacturonase was extracted from Chrysophyllum albidum fruit and partially purified using ammonium sulphate precipitation (80% saturation), dialysis, and gel filtration (Sephadex-G 100). Protein content and polygalacturonase activity were assayed, and the effects of pH, temperature, and substrate concentration on the enzyme activity were determined. The protein concentration and polygalacturonase activity for the ripe fruit were 1.35 mg/mL and 76.35 U/mg protein, respectively, while for the unripe fruit, it was 0.925 mg/mL and 1.59 U/mg protein. Upon partial purification, five fractions (fraction 18, 21-24) had the highest polygalacturonase activity. The optimum pH and temperature for Chrysophyllum albidum juice extract were 4.5 and 40°C, respectively. The enzyme activity increased with an increase in substrate concentration. The Vmax for polygalacturonase was 4.42 U/mg protein, and Km was 1.38 mg/mL. In conclusion, Chrysophyllum albidum fruit is a source of polygalacturonase which could be explored.
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Allen RL and Lonsdale DM. Molecular characterisation of one of the maize polygalacturonase gene family members, which are expressed during late pollen development. Plant J. 1993; 3(2):261-271.
Buchanan BB, Gruissem W, Jones RL. Biochemistry and molecular biology of plants. (2nd ed.). West Sussex, UK: Wiley Blackwell; 2015. 80-90 p.
Nakagawa T, Taniguchi S, Miyaji T, Tomizuka N. Isolation and characterisation of psychrophilic yeast producing coldadapted pectinolytic enzymes. Lett Appl Microbiol. 2004; 38(5):383-387.
Pedrolli DB, Monteiro AC, Gomes E, Carmona EC. Pectin and pectinases: production, characterisation and industrial application of microbial pectinolytic enzymes. Open Biotechnol J. 2009; 3:9-18.
Silva D, Da Silva-Martins E, Da Silva R, Gmes E. Pectinase production by Penicillium viridicatum RFC3 by solid states fermentation using agricultural wastes and agro-industrial by products. Braz J Microbiol. 2002; 33(4):1-10.
Souza J, Silva E, Maia M, Teixeira M. Screening of fungal strains for pectinolytic activity: Endopolygalacturonase production of Peacilomyces clavisporus 2A, UMIDA. 1. Proc Biochem. 2003; 39(4):455-458.
Kashyap DR, Vohra PK, Chopra S, Tewari R. Applications of pectinases in the commercial sector: A review. Bioresour Technol. 2001;77(3):215-227.
Arunachalam C and Asha S. Pectinolytic enzyme-A review of new studies. Biotechnol Adv. 2010; 561:1-5.
Avwioroko OJ, Anigboro AA, Atanu FO, Otuechere CA, Alfred MO, Abugo JN, Omorogie MO. Investigation of the binding interaction of α-amylase with Chrysophyllum albidum seed extract and its silver nanoparticles: a multispectroscopic approach. Chem Data Coll. 2020; 29:100517.
Ajayi AM, Chidebe EO, Ben-Azu B, Umukoro S. Chrysophyllum albidum (African star apple) fruitsupplemented diet enhances cognitive functions and attenuates lipopolysaccharide-induced memory impairment, oxidative stress, and release of proinflammatory cytokines.
Nutr. 2020; 45(2):1-13.
Amin F, Mohsin A, Bhatti HN, Bilal M. Production, thermodynamic characterisation, and fruit juice quality improvement characteristics of an Exo-polygalacturonase from Penicillium janczewskii. Biochim Biophys Acta Proteins Proteomics. 2020; 1868(5): 140379.
Aslam F, Ansari A, Aman A, Baloch G, Nisar G, Baloch AH, Rehman HU. Production of commercially important enzymes from Bacillus licheniformis KIBGE-IB3 using date fruit wastes as substrate. J Genet Eng Biotechnol. 2020; 18(1):1-7.
Dasari PK. Parametric optimisations for pectinase production by Aspergillus awamori. GSC Biol Pharm Sci. 2020; 12(2):93-98.
Gautam RL and Naraian R. Trichoderma, a factory of multipurpose enzymes: Cloning of enzymatic genes. In: Hesham A, Upadhyay R, Sharma G, Manoharachary C, Gupta V, eds. Fungal Biotechnology and Bioengineering. Cham, Switzerland: Springer; 2020. 137-162 p.
Yadav A, Ali AAM, Ingawale M, Raychaudhuri S, Gantayet LM, Pandit A. Enhanced co-production of pectinase, cellulase and xylanase enzymes from Bacillus subtilis ABDR01 upon ultrasonic irradiation. Proc Biochem. 2020; 92:197-201.
Akhter N, Morshed MA, Uddin A, Begum F, Sultan T, Azad AK. Production of pectinase by Aspergillus niger cultured in solid state media. Int J Biosci. 2011; 1(1):33-42.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem. 1951;193(1):265-275.
Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem. 1959; 31(3):426-8.
Chinedu SN, Dayo-Odukoya OP, Iheagwam FN. Partial purification and kinetic properties of polygalacturonase from Solanum macrocarpum L. fruit. Biotechnol Adv. 2017; 16(1):27-33.
Ázar RIL, da Luz Morales M, Maitan-Alfenas GP, Falkoski DL, Alfenas RF, Guimarães VM. Apple juice clarification by a purified polygalacturonase from Calonectria pteridis. Food Bioprod Proc. 2020; 119:238-245.
Cheng Z, Xian L, Chen D, Lu J, Wei Y, Du L, Wang Q, Chen Y, Lu B, Bi D, Zhang Z. Development of an innovative process for high-temperature fruit juice extraction using a novel thermophilic endopolygalacturonase from Penicillium oxalicum. Front Microbiol. 2020; 11:Article 1200.
Karaoğlan M and Erden-Karaoğlan F. Effect of codon optimisation and promoter choice on recombinant endopolygalacturonase production in Pichia pastoris. Enzyme Microb Technol. 2020; 139:109589.
Aminzadeh S, Naderi-Manesh H, Khajeh K, NaderiManesh M. Purification, characterisation, kinetic properties, and thermal behavior of extracellular polygalacturonase produced by filamentous fungus Tetracoccosporium sp. Appl Biochem Biotechnol. 2006; 135(3):193-208.