Polyamide Microparticles with Immobilized Enological Pectinase As Efficient Biocatalysts for Wine Clarification: The Role of the Polymer Support

Overview

Journal Molecules

Publisher MDPI

Date 2025 Jan 11

PMID 39795171

Authors

Sandra C Oliveira

Samuel M Araujo

Nadya V Dencheva

Zlatan Z Denchev

Affiliations

Soon will be listed here.

Abstract

Free pectinase is commonly employed as a biocatalyst in wine clarification; however, its removal, recovery, and reuse are not feasible. To address these limitations, this study focuses on the immobilization of a commercial pectinolytic preparation (Pec) onto highly porous polymer microparticles (MPs). Seven microparticulate polyamide (PA) supports, namely PA4, PA6, PA12 (with and without magnetic properties), and the copolymeric PA612 MP, were synthesized through activated anionic ring-opening polymerization of various lactams. Pectinase was non-covalently immobilized on these supports by adsorption, forming Pec@PA conjugates. Comparative activity and kinetic studies revealed that the Pec@PA12 conjugate exhibited more than twice the catalytic efficiency of the free enzyme, followed by Pec@PA6-Fe and Pec@PA4-Fe. All Pec@PA complexes were tested in the clarification of industrial rosé must, demonstrating similar or better performance compared to the free enzyme. Some immobilized biocatalysts supported up to seven consecutive reuse cycles, maintaining up to 50% of their initial activity and achieving complete clarification within 3-30 h across three consecutive cycles of application. These findings highlight the potential for industrial applications of noncovalently immobilized pectinase on various polyamide microparticles, with possibilities for customization of the conjugates' properties.

References

Kassara S, Li S, Smith P, Blando F, Bindon K . Pectolytic enzyme reduces the concentration of colloidal particles in wine due to changes in polysaccharide structure and aggregation properties. Int J Biol Macromol. 2019; 140:546-555. DOI: 10.1016/j.ijbiomac.2019.08.043. View

Yushkova E, Nazarova E, Matyuhina A, Noskova A, Shavronskaya D, Vinogradov V . Application of Immobilized Enzymes in Food Industry. J Agric Food Chem. 2019; 67(42):11553-11567. DOI: 10.1021/acs.jafc.9b04385. View

Jiang X, Lu Y, Liu S . Effects of Different Yeasts on Physicochemical and Oenological Properties of Red Dragon Fruit Wine Fermented with , and . Microorganisms. 2020; 8(3). PMC: 7142936. DOI: 10.3390/microorganisms8030315. View

Behram T, Pervez S, Nawaz M, Ahmad S, Jan A, Rehman H . Development of Pectinase Based Nanocatalyst by Immobilization of Pectinase on Magnetic Iron Oxide Nanoparticles Using Glutaraldehyde as Crosslinking Agent. Molecules. 2023; 28(1). PMC: 9823745. DOI: 10.3390/molecules28010404. View

Farago P, Galatus R, Hintea S, Bosca A, Feurdean C, Ilea A . An Intra-Oral Optical Sensor for the Real-Time Identification and Assessment of Wine Intake. Sensors (Basel). 2019; 19(21). PMC: 6864861. DOI: 10.3390/s19214719. View

Espejo F . Role of commercial enzymes in wine production: a critical review of recent research. J Food Sci Technol. 2021; 58(1):9-21. PMC: 7813895. DOI: 10.1007/s13197-020-04489-0. View

Edwards V . The influence of high substrate concentrations on microbial kinetics. Biotechnol Bioeng. 1970; 12(5):679-712. DOI: 10.1002/bit.260120504. View

Miao Q, Zhang C, Zhou S, Meng L, Huang L, Ni Y . Immobilization and Characterization of Pectinase onto the Cationic Polystyrene Resin. ACS Omega. 2021; 6(47):31683-31688. PMC: 8637955. DOI: 10.1021/acsomega.1c04374. View

Rexova-Benkova L, Omelkova J, Veruovic B, Kubanek V . Mode of action of endopolygalacturonase immobilized by adsorption and by covalent binding via amino groups. Biotechnol Bioeng. 1989; 34(1):79-85. DOI: 10.1002/bit.260340111. View

10.

Barbosa O, Ortiz C, Berenguer-Murcia A, Torres R, Rodrigues R, Fernandez-Lafuente R . Strategies for the one-step immobilization-purification of enzymes as industrial biocatalysts. Biotechnol Adv. 2015; 33(5):435-56. DOI: 10.1016/j.biotechadv.2015.03.006. View

11.

Rehman H, Nawaz M, Pervez S, Jamal M, Attaullah M, Aman A . Encapsulation of pectinase within polyacrylamide gel: characterization of its catalytic properties for continuous industrial uses. Heliyon. 2020; 6(8):e04578. PMC: 7453123. DOI: 10.1016/j.heliyon.2020.e04578. View

12.

Pretorius I . Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking. Yeast. 2000; 16(8):675-729. DOI: 10.1002/1097-0061(20000615)16:8<675::AID-YEA585>3.0.CO;2-B. View

13.

Benucci I, Lombardelli C, Liburdi K, Acciaro G, Zappino M, Esti M . Immobilised native plant cysteine proteases: packed-bed reactor for white wine protein stabilisation. J Food Sci Technol. 2016; 53(2):1130-9. PMC: 4837734. DOI: 10.1007/s13197-015-2125-4. View

14.

Ottone C, Romero O, Aburto C, Illanes A, Wilson L . Biocatalysis in the winemaking industry: Challenges and opportunities for immobilized enzymes. Compr Rev Food Sci Food Saf. 2020; 19(2):595-621. DOI: 10.1111/1541-4337.12538. View

15.

Saxena S, Shukla S, Thakur A, Gupta R . Immobilization of polygalacturonase from Aspergillus niger onto activated polyethylene and its application in apple juice clarification. Acta Microbiol Immunol Hung. 2008; 55(1):33-51. DOI: 10.1556/AMicr.55.2008.1.3. View

16.

Diano N, Grimaldi T, Bianco M, Rossi S, Gabrovska K, Yordanova G . Apple juice clarification by immobilized pectolytic enzymes in packed or fluidized bed reactors. J Agric Food Chem. 2008; 56(23):11471-7. DOI: 10.1021/jf8019437. View

17.

Hong M, Chen E . Completely recyclable biopolymers with linear and cyclic topologies via ring-opening polymerization of γ-butyrolactone. Nat Chem. 2015; 8(1):42-9. DOI: 10.1038/nchem.2391. View

18.

Oliveira S, Dencheva N, Denchev Z . Immobilization of Enological Pectinase on Magnetic Sensitive Polyamide Microparticles for Wine Clarification. Foods. 2024; 13(3). PMC: 10855548. DOI: 10.3390/foods13030420. View