Abstract
The development of biotechnological approaches for preventing chill haze formation has attracted great interest in brewing research. The current work provides an innovative biocatalytic system, based on immobilized tannase (as phenolic-degrading enzyme), for the continuous treatment of wort in fluidized-bed reactor (FBR). The covalent immobilization on chitosan beads has been performed using a food-grade cross-linker. The initial protein concentration of 1.35 mgBSAeq/mL allowed us to maximize the specific activity of the biocatalyst (0.017 I.U./mgBSAeq), which was characterized by a higher pH and storage stability than that of the free enzyme. The continuous treatment in FBR has been optimized varying the flow rate (Qv) and the amount of biocatalyst, and the suitable conditions for the continuous treatment of synthetic wort were 560 mL/min (Qv) and 5.0 g of biocatalyst. Immobilized tannase exhibited excellent operational stability in FBR and has been reused eight times retaining 60% of its initial activity. The continuous and specific haze-preventing biotechnological treatment provided in this study, and based on food-grade immobilized tannase, may be successfully applied during the phase of post-mashing, when the operating conditions (T = 40 °C, pH = 5) match those optimal for the catalytic activity of the enzyme.
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The data presented in this study are available upon request from the corresponding author.
References
Dhiman S, Mukherjee G, Kumar A et al (2017) Fungal tannase: recent advances and industrial applications. In: Developments in fungal biology and applied mycology. Springer, Singapore, pp 295–313
Van Diepeningen AD, Debets AJM, Varga J, Van Der Gaag M, Swart K, Hoekstra RF (2004) Efficient degradation of tannic acid by black Aspergillus species. Mycol Res 108:919–925. https://doi.org/10.1017/S0953756204000747
Lu MJ, Chu SC, Yan L, Chen C (2009) Effect of tannase treatment on protein-tannin aggregation and sensory attributes of green tea infusion. LWT 42:338–342. https://doi.org/10.1016/j.lwt.2008.05.015
Su E, Xia T, Gao L, Dai Q, Zhang Z (2009) Immobilization and characterization of tannase and its haze-removing. Food Sci Technol Int 15:545–552. https://doi.org/10.1177/1082013209352919
Cao QQ, Zou C, Zhang YH, Du QZ, Yin JF, Shi J, Xu YQ (2019) Improving the taste of autumn green tea with tannase. Food Chem 277:432–437. https://doi.org/10.1016/j.foodchem.2018.10.146
Shao Y, Zhang YH, Zhang F, Yang QM, Weng HF, Xiao Q, Xiao AF (2020) Thermostable tannase from Aspergillus niger and its application in the enzymatic extraction of green tea. Molecules 25:952. https://doi.org/10.3390/molecules25040952
Rout S, Banerjee R (2006) Production of tannase under mSSF and its application in fruit juice debittering
Kumar SJ, Kumar V (2021) Effect of immobilized enzymes naringinase and tannase produced from Aspergillus sp. Isolate Mk156394 isolated from rotten pomelo on quality characteristics of citrus limetta juice and process optimization by using response surface methodology. Biointerface Res Appl Chem 11:9646–9657. https://doi.org/10.33263/BRIAC112.96469657
Benucci I, Mazzocchi C, Lombardelli C, Esti M (2023) Phenolic-degrading enzymes: effect on haze active phenols and chill haze in India pale ale beer. Foods 12:77. https://doi.org/10.3390/foods12010077
Schwedt G, Stein K (1994) Immobilized enzymes as tools in food analysis. Einsatz immobilisierter Enzyme in der Lebensmittelanalytik 199:171–182
Wang C, Chen PX, Xiao Q, Yang QM, Weng HF, Zhang YH, Xiao AF (2021) Chitosan activated with genipin: a nontoxic natural carrier for tannase immobilization and its application in enhancing biological activities of tea extract. Mar Drugs 19:66. https://doi.org/10.3390/md19030166
Wu C, Xu C, Ni H, Yang Q, Cai H, Xiao A (2016) Preparation and characterization of tannase immobilized onto carboxyl-functionalized superparamagnetic ferroferric oxide nanoparticles. Bioresour Technol 205:67–74. https://doi.org/10.1016/j.biortech.2016.01.032
Jana A, Halder SK, Ghosh K, Paul T, Vágvölgyi C, Mondal KC, Mohapatra PKD (2015) Tannase immobilization by chitin-alginate based adsorption-entrapment technique and its exploitation in fruit juice clarification. Food Bioproc Tech 8:2319–2329. https://doi.org/10.1007/s11947-015-1586-9
Flores EEE, Cardoso FD, Siqueira LB, Ricardi NC, Costa TH, Rodrigues RC, Hertz PF (2019) Influence of reaction parameters in the polymerization between genipin and chitosan for enzyme immobilization. Process Biochem 84:73–80. https://doi.org/10.1016/j.procbio.2019.06.001
Benucci I, Mazzocchi C, Lombardelli C, Cacciotti I, Esti M (2019) Multi-enzymatic systems immobilized on chitosan beads for pomegranate juice treatment in fluidized bed reactor: effect on haze-active molecules and chromatic properties. Food Bioproc Tech 12:1559–1572. https://doi.org/10.1007/s11947-019-02315-w
Cacciotti I, Lombardelli C, Benucci I, Esti M (2019) Clay/chitosan biocomposite systems as novel green carriers for covalent immobilization of food enzymes. J Mater Res Technol 8:3644–3652. https://doi.org/10.1016/j.jmrt.2019.06.002
Benucci I, Lombardelli C, Liburdi K, Acciaro G, Zappino M, Esti M (2016) Immobilised native plant cysteine proteases: packed-bed reactor for white wine protein stabilisation. J Food Sci Technol 53:1130–1139. https://doi.org/10.1007/s13197-015-2125-4
Benucci I, Lombardelli C, Cacciotti I, Liburdi K, Nanni F, Esti M (2016) Chitosan beads from microbial and animal sources as enzyme supports for wine application. Food Hydrocoll 61:191–200. https://doi.org/10.1016/j.foodhyd.2016.05.016
Benucci I, Lombardelli C, Cacciotti I, Esti M (2020) Papain covalently immobilized on chitosan-clay nanocomposite films: application in synthetic and real white wine. Nanomaterials 10:1622. https://doi.org/10.3390/nano10091622
Benucci I, Caso MC, Bavaro T, Masci S, Keršienė M, Esti M (2020) Prolyl endopeptidase from Aspergillus niger immobilized on a food-grade carrier for the production of gluten-reduced beer. Food Control 110:106987. https://doi.org/10.1016/j.foodcont.2019.106987
Priyadarshi R, Rhim JW (2020) Chitosan-based biodegradable functional films for food packaging applications. Innov Food Sci Emerg Technol 62:102346
Yang X, Chen Y, Yao S, Qian J, Guo H, Cai X (2019) Preparation of immobilized lipase on magnetic nanoparticles dialdehyde starch. Carbohydr Polym 218:324–332. https://doi.org/10.1016/j.carbpol.2019.05.012
Tan L, Li LQ, Dong J, Liu ZL, Liu YP, Lu M (2015) Immobilization of papain on flexible magnetic nanoparticles. Appl Mech Mater 723:511–514. https://doi.org/10.4028/www.scientific.net/AMM.723.511
Chen H, Liu L, Lv S, Liu X, Wang M, Song A, Jia X (2010) Immobilization of Aspergillus niger xylanase on chitosan using dialdehyde starch as a coupling agent. Appl Biochem Biotechnol 162:24–32. https://doi.org/10.1007/s12010-009-8790-x
Cappannella E, Benucci I, Lombardelli C, Liburdi K, Bavaro T, Esti M (2016) Immobilized lysozyme for the continuous lysis of lactic bacteria in wine: bench-scale fluidized-bed reactor study. Food Chem 210:49–55. https://doi.org/10.1016/j.foodchem.2016.04.089
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Zhang DH, Yuwen LX, Peng LJ (2013) Parameters affecting the performance of immobilized enzyme. J Chem 2013:1–7
Zhang DH, Yuwen LX, Li C, Li YQ (2012) Effect of poly(vinyl acetate-acrylamide) microspheres properties and steric hindrance on the immobilization of Candida rugosa lipase. Bioresour Technol 124:233–236. https://doi.org/10.1016/j.biortech.2012.08.083
Li R, Fu G, Liu C, McClements DJ, Wan Y, Wang S, Liu T (2018) Tannase immobilisation by amino-functionalised magnetic Fe3O4-chitosan nanoparticles and its application in tea infusion. Int J Biol Macromol 114:1134–1143. https://doi.org/10.1016/j.ijbiomac.2018.03.077
Kumar S, Beniwal V, Kumar N, Kumar A, Chhokar V, Khaket TP (2015) Biochemical characterization of immobilized tannase from Aspergillus awamori. Biocatal Agric Biotechnol 4:398–403. https://doi.org/10.1016/j.bcab.2015.07.004
Abdel-Naby MA, Sherif AA, El-Tanash AB, Mankarios AT (1999) Immobilization of Aspergillus oryzae tannase and properties of the immobilized enzyme. J Appl Microbiol 87:108–114. https://doi.org/10.1046/j.1365-2672.1999.00799.x
Ong CB, Annuar MS (2018) Immobilization of cross-linked tannase enzyme on multiwalled carbon nanotubes and its catalytic behavior. Prep Biochem Biotechnol 48:181–187. https://doi.org/10.1080/10826068.2018.1425707
Alias MR, Ong CB, Annuar MSM (2023) Adsorption of tannase from Aspergillus ficuum to carboxyl-functionalized multi-walled carbon nanotubes. J Serb Chem Soc. https://doi.org/10.2298/JSC221121009A
Sharma S, Agarwal L, Saxena RK (2008) Purification, immobilization and characterization of tannase from Penicillium variable. Bioresour Technol 99:2544–2551. https://doi.org/10.1016/j.biortech.2007.04.035
Yu X, Li Y, Wang C, Wu D (2004) Immobilization of Aspergillus niger tannase by microencapsulation and its kinetic characteristics. Biotechnol Appl Biochem 40:151. https://doi.org/10.1042/ba20030180
El-Tanash AB, Sherief AA, Nour A (2011) Catalytic properties of immobilized tannase produced from Aspergillus aculeatus compared with the free enzyme. Braz J Chem Eng 28:381–391
Wang SN, Zhang CR, Qi BK, Sui XN, Jiang LZ, Li Y, Zhang QZ (2014) Immobilized alcalase alkaline protease on the magnetic chitosan nanoparticles used for soy protein isolate hydrolysis. Eur Food Res Technol 239:1051–1059. https://doi.org/10.1007/s00217-014-2301-1
Sharma S, Krishan Bhat T, Gupta MN (2002) Bioaffinity immobilization of tannase from Aspergillus niger on concanavalin A-Sepharose CL-4B. Biotechnol Appl Biochem 35:165–169. https://doi.org/10.1111/j.1470-8744.2002.tb01185.x
Dutta N, Miraz SM, Khan MU, Karekar SC, Usman M, Khan SM, Thiruvengadam M (2021) Heterologous expression and biophysical characterization of a mesophilic tannase following manganese nanoparticle immobilization. Colloids Surf B Biointerfaces 207:112011. https://doi.org/10.1016/j.colsurfb.2021.112011
Ching CB, Ho YY (1984) Flow dynamics of immobilized enzyme reactors. Appl Microbiol Biotechnol 20:303–309
Liu J, Chen G, Yan B, Yi W, Yao J (2022) Biodiesel production in a magnetically fluidized bed reactor using whole-cell biocatalysts immobilized within ferroferric oxide-polyvinyl alcohol composite beads. Bioresour Technol 355:127253. https://doi.org/10.1016/j.biortech.2022.127253
Zhou GX, Chen GY, Yan BB (2014) Biodiesel production in a magnetically-stabilized, fluidized bed reactor with an immobilized lipase in magnetic chitosan microspheres. Biotechnol Lett 36:63–68. https://doi.org/10.1007/s10529-013-1336-x
Ricca E, Calabrò V, Curcio S, Basso A, Gardossi L, Iorio G (2010) Fructose production by inulinase covalently immobilized on Sepabeads in batch and fluidized bed bioreactor. Int J Mol Sci 11:1180–1189. https://doi.org/10.3390/ijms11031180
Aharwar A, Parihar DK (2023) Talaromyces verruculosus tannase immobilization, characterization, and application in tea infusion treatment. Biomass Convers Biorefin 13:261–272. https://doi.org/10.1007/s13399-020-01162-6
Hota SK, Dutta JR, Banerjee R (2007) Immobilization of tannase from Rhizopus oryzae and its efficiency to produce gallic acid from tannin rich agro-residues
Schons PF, Lopes FCR, Battestin V, MacEdo GA (2011) Immobilization of Paecilomyces variotii tannase and properties of the immobilized enzyme. J Microencapsul 28:211–219. https://doi.org/10.3109/02652048.2011.552988
Acknowledgements
This work was supported by StaBirVino project “Enzimi immobilizzati per la stabilizzazione sostenibile di birra e vino” (Grant A0375-2020-36649) funded by Lazio Region (Italy) in the context of Progetti Gruppi di Ricerca, LazioInnova 2020-2022.
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IB, ME: conceptualization, validation, data curation, writing—review and editing, visualization, project administration. IB, CL: methodology, formal analysis, investigation, resources, writing—original draft. ME: funding acquisition.
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Benucci, I., Lombardelli, C. & Esti, M. Innovative continuous biocatalytic system based on immobilized tannase: possible prospects for the haze-active phenols hydrolysis in brewing industry. Eur Food Res Technol 249, 2625–2633 (2023). https://doi.org/10.1007/s00217-023-04323-9
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DOI: https://doi.org/10.1007/s00217-023-04323-9