Abstract
A novel thermostable caffeine dehydrogenase enzyme was isolated from Pichia manshurica strain CD1. At increased temperatures, the enzyme caffeine dehydrogenase exhibits increased activity. The enzyme also exhibits high stability after the heat treatment and remains up to 56% stable at 100 ºC compared to its untreated condition. A Comprehensive analysis was carried out to monitor the thermodynamic parameters (Ea, ΔG, ΔH, and ΔS) of the enzyme. In this study, the conformational changes of caffeine dehydrogenase in native and thermally treated states were investigated by CD spectroscopy and intrinsic tryptophan fluorescence spectroscopy. The far-UV CD spectra results showed that the protein was able to maintain its alpha-helical structure after the heat treatment and the near-UV CD spectra results showed that temperature had no such effect on the protein’s tertiary structure. The results of tryptophan fluorescence spectroscopy of the enzyme showed a temperature-dependent dynamic quenching. The fluorescence intensity of the protein was decreased as the degree of freedom in the tryptophan- region of the protein was increased with temperature. Thus, the enzyme does not undergo denaturation at high temperatures and can maintain its conformation to remain catalytically active. Therefore, it can be concluded that the thermal stability of caffeine dehydrogenase is an essential key factor in increasing commercial interest due to its high catalytic efficiency and specificity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Change history
14 July 2023
A Correction to this paper has been published: https://doi.org/10.1007/s11756-023-01477-5
References
Afzal AJ, Ali S, Latif F, Rojoka MI, Siddiqui KS (2005) Innovative kinetic and thermodynamic analysis of a purified superactive xylanase from Scopulariopsis sp. Appl Biochem Biotechnol 120:51–70. https://doi.org/10.1385/abab:120:1:51
Arndt C, Koristka S, Bartsch H, Bachmann M (2012) Native polyacrylamide gels. Methods Mol Biol 869:49–53. https://doi.org/10.1007/978-1-61779-821-4_5
Bhatt K, Lal S, Srinivasan R, Joshi B (2020) Molecular analysis of Bacillus velezensis KB 2216, purification and biochemical characterization of alpha-amylase. Int J Biol Macromol 164:3332–3339. https://doi.org/10.1016/j.ijbiomac.2020.08.205
Bushueva TL, Busel EP, Burstein EA (1978) Relationship of thermal quenching of protein fluorescence to intramolecular structural mobility. Biochim Biophys Acta - Protein Struct 534:141–152. https://doi.org/10.1016/0005-2795(78)90484-1
Chafik A, Essamadi A, Çelik SY, Mavi A (2020) A novel acid phosphatase from cactus (Opuntia megacantha Salm-Dyck) cladodes: purification and biochemical characterization of the enzyme. Int J Biol Macromol 160:991–999. https://doi.org/10.1016/j.ijbiomac.2020.05.175
Chen YH, Huang YH, Wen CC, Wang YH, Chen WL, Chen LC, Tsay HJ (2008) Movement disorder and neuromuscular change in zebrafish embryos after exposure to caffeine. Neurotoxicol Teratol 30:440–447. https://doi.org/10.1016/j.ntt.2008.04.003
Chevallet M, Luche S, Rabilloud T (2006) Silver staining of proteins in polyacrylamide gels. Nat Protoc 1. https://doi.org/10.1038/nprot.2006.288
Collins T, Gerday C, Feller G (2005) Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev 29:3–23. https://doi.org/10.1016/j.femsre.2004.06.005
Dash SS, Gummadi SN (2006) Catabolic pathways and biotechnological applications of microbial caffeine degradation. Biotechnol Lett 28:1993–2002. https://doi.org/10.1007/s10529-006-9196-2
Fernandes AS, Mello FVC, Thode Filho S, Carpes RM, Honorio JG, Marques MRC, Felzenszwalb I, Ferraz ERA (2017) Impacts of discarded coffee waste on human and environmental health. Ecotoxicol Environ Saf 141:30–36. https://doi.org/10.1016/j.ecoenv.2017.03.011
Gentile K, Bhide A, Kauffman J, Ghosh S, Maiti S, Adair J, Lee TH, Sen A (2021) Enzyme aggregation and fragmentation induced by catalysis relevant species. Phys Chem Chem Phys 23:20709–20717. https://doi.org/10.1039/d1cp02966e
Greenfield NJ (2006) Using circular dichroism spectra to estimate protein secondary structure. Nat Protoc 1:2876–2890. https://doi.org/10.1038/nprot.2006.202
Gu X, Zhang S, Ma W, Wang Q, Li Y, Xia C, Xu Y, Zhang T, Yang L, Zhou M (2022) The impact of instant coffee and decaffeinated coffee on the gut microbiota and depression-like behaviors of sleep-deprived rats. Front Microbiol 13:778512. https://doi.org/10.3389/fmicb.2022.778512
Gummadi SN, Bhavya B, Ashok N (2011) Physiology, biochemistry and possible applications of microbial caffeine degradation. Appl Microbiol Biotechnol 93(2):545–554. https://doi.org/10.1007/s00253-011-3737-x
Gutarra MLE, Godoy MG, Maugeri F, Rodrigues MI, Freire DMG, Castilho LR (2009) Production of an acidic and thermostable lipase of the mesophilic fungus Penicillium simplicissimum by solid-state fermentation. Bioresour Technol 100:5249–5254. https://doi.org/10.1016/j.biortech.2008.08.050
He S, Qiao X, Zhang S, Xia J, Wang L, Liu S (2023) Urate oxidase from tea microbe Colletotrichum camelliae is involved in the caffeine metabolism pathway and plays a role in fungal virulence. Front Nutr 9:1038806. https://doi.org/10.3389/fnut.2022.1038806
Heckman MA, Weil J, De Mejia EG (2010) Caffeine (1, 3, 7-trimethylxanthine) in foods: a comprehensive review on consumption, functionality, safety, and regulatory matters. J Food Sci 75:R77–R87. https://doi.org/10.1111/j.1750-3841.2010.01561.x
Jaiswal A, Preet M, Tripti B (2017) Production and optimization of lipase enzyme from mesophiles and thermophiles. J Microb Biochem Technol 9:126–131. https://doi.org/10.4172/1948-5948.1000355
Jayanthi N, Purwanto MGM, Chrisnasari R, Pantjajani T, Wahjudi A, Sugiarto M (2019) Characterization of thermostable chitinase from Bacillus licheniformis B2. IOP Conf Ser : Earth Environ Sci 293:012030. https://doi.org/10.1088/1755-1315/293/1/012030
Karray A, Alonazi M, Horchani H, Bacha AB (2021) A novel thermostable and alkaline protease produced from Bacillus stearothermophilus isolated from olive oil mill sols suitable to industrial biotechnology. Mol 26:1139. https://doi.org/10.3390/molecules26041139
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685. https://doi.org/10.1038/227680a0
Lovallo WR, Whitsett TL, Al’Absi M (2005) Caffeine stimulation of cortisol secretion across the waking hours in relation to caffeine intake levels. Psychosom Med 67:734–739. https://doi.org/10.1097/01.psy.0000181270.20036.06
Madyastha KM, Sridhar GR, Vadiraja BB, Madhavi YS (1999) Purification and partial characterization of caffeine oxidase—a novel enzyme from a mixed culture consortium. Biochem Biophys Res Commun 263:460–464. https://doi.org/10.1006/bbrc.1999.1401
Micsonai A, Wien F, Kernya L (2015) Accurate secondary structure prediction and fold recognition for circular dichroism spectroscopy. Proc Natl Acad Sci U S A 112:E3095–E3103. https://doi.org/10.1073/pnas.1500851112
Micsonai A, Bulyáki É, Kardos J (2021) BeStSel: from secondary structure analysis to protein fold prediction by circular dichroism spectroscopy. Methods Mol Biol (Clifton N J) 2199:175–189. https://doi.org/10.1007/978-1-0716-0892-0_11
Mohanty SK, Yu CL, Das S, Louie TM, Gakhar L, Subramanian M (2012) Delineation of the caffeine C-8 oxidation pathway in Pseudomonas sp. strain CBB1 via characterization of a new trimethyluric acid monooxygenase and genes involved in trimethyluric acid metabolism. J Bacteriol 194:3872–3882. https://doi.org/10.1128/jb.00597-12
Mohapatra BR, Harris N, Nordin R, Mazumder A (2006) Purification and characterization of a novel caffeine oxidase from Alcaligenes species. J Biotechnol 125:319–327. https://doi.org/10.1016/j.jbiotec.2006.03.018
Möller M, Denicola A (2002) Laboratory exercises protein tryptophan accessibility studied by fluorescence quenching. Biochem Mol Biol Educ 30:175–178. https://doi.org/10.1002/bmb.2002.494030030035
Murtaza A, Muhammad Z, Iqbal A, Ramzan R, Liu Y, Pan S, Hu W (2018) Aggregation and conformational changes in native and thermally treated polyphenol oxidase from apple juice (Malus domestica). Front Chem 6:203. https://doi.org/10.3389/fchem.2018.00203
Okpara MO, Okpara MO (2022) Microbial enzymes and their applications in food industry: a mini-review. Adv Enzym Res 10:23–47. https://doi.org/10.4236/aer.2022.101002
Parvin R, Bhattacharya S, Chaudhury SS, Roy U, Mukherjee J, Gachhui R (2023) Production, purification and characterization of a novel thermostable caffeine dehydrogenase from Pichia manshurica strain CD1 isolated from kombucha tea. Microbiol (Russian Fed) 92:230–241. https://doi.org/10.1134/s0026261722601476
Retamal CA, Thiebaut P, Alves EW (1999) Protein purification from polyacrylamide gels by sonication extraction. Anal Biochem 268:15–20. https://doi.org/10.1006/abio.1998.2977
Reyes CM, Cornelis MC (2018) Caffeine in the diet: country-level consumption and guidelines. Nutr 10:1772. https://doi.org/10.3390/nu10111772
Saldaña MDA, Zetzl C, Mohamed RS, Brunner G (2002) Decaffeination of guaraná seeds in a microextraction column using water-saturated CO2. J Supercrit Fluids 22:119–127. https://doi.org/10.1016/s0896-8446(01)00121-8
Shanmugam MK, Rathinavelu S, Gummadi SN (2021) Self-directing optimization for enhanced caffeine degradation in synthetic coffee wastewater using induced cells of Pseudomonas sp.: bioreactor studies. J Water Process Eng 44:102341. https://doi.org/10.1016/j.jwpe.2021.102341
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. Mol 25:952. https://doi.org/10.3390/molecules25040952
Sharif R, Ahmad SW, Anjum H, Ramzan N, Malik SR (2014) Effect of infusion time and temperature on decaffeination of tea using liquid–liquid extraction technique. J Food Process Eng 37:46–52. https://doi.org/10.1111/jfpe.12058
Sharma S, Vaid S, Bhat B, Singh S, Bajaj BK (2019) Thermostable enzymes for industrial biotechnology. Adv Enzym Technol 1st ed. pp 469–495. https://doi.org/10.1016/b978-0-444-64114-4.00017-0
Silva JDC, de França PRL, Converti A, Porto TS (2019) Pectin hydrolysis in cashew apple juice by aspergillus aculeatus URM4953 polygalacturonase covalently-immobilized on calcium alginate beads: a kinetic and thermodynamic study. Int J Biol Macromol 126:820–827. https://doi.org/10.1016/j.ijbiomac.2018.12.236
Soares da Silva O, Lira de Oliveira R, de Carvalho Silva J, Converti A, Porto TS (2018) Thermodynamic investigation of an alkaline protease from aspergillus tamarii URM4634: a comparative approach between crude extract and purified enzyme. Int J Biol Macromol 109:1039–1044. https://doi.org/10.1016/j.ijbiomac.2017.11.081
Summers RM, Mohanty SK, Gopishetty S, Subramanian M (2015) Genetic characterization of caffeine degradation by bacteria and its potential applications. Microb Biotechnol 8:369–378. https://doi.org/10.1111/1751-7915.12262
Suresh PV, Anil Kumar PK (2012) Enhanced degradation of α-chitin materials prepared from shrimp processing byproduct and production of N-acetyl-d-glucosamine by thermoactive chitinases from soil mesophilic fungi. Biodegradation 23:597–607. https://doi.org/10.1007/s10532-012-9536-y
Temsaah HR, Azmy AF, Raslan M, Ahmed AE, Hozayen WG (2018) Isolation and characterization of thermophilic enzymes producing microorganisms for potential therapeutic and industrial use. J Pure Appl Microbiol 12:1687–1702. https://doi.org/10.22207/jpam.12.4.02
Vaseekaran S, Balakumar S, Arasaratnam V (2010) Isolation and identification of a bacterial strain producing thermostable α- amylase. Trop Agric Res 22:1–11. https://doi.org/10.4038/tar.v22i1.2603
Yu CL, Kale Y, Gopishetty S, Louie TM, Subramanian M (2008) A novel caffeine dehydrogenase in Pseudomonas sp. strain CBB1 oxidizes caffeine to trimethyluric acid. J Bacteriol 190:772–776. https://doi.org/10.1128/jb.01390-07
Zabot GL (2020) Decaffeination using supercritical carbon dioxide. Green Sustain Process Chem Environ Eng Sci Supercrit Carbon Dioxide as Green Solvent 255–278. https://doi.org/10.1016/b978-0-12-817388-6.00011-8
Zhou B, Ma C, Wang H, Xia T (2018) Biodegradation of caffeine by whole cells of tea-derived fungi Aspergillus sydowii, Aspergillus niger and optimization for caffeine degradation. BMC Microbiol 18:1–10. https://doi.org/10.1186/s12866-018-1194-8
Acknowledgements
We thank Prof. Utpal Chandra Chaudhuri for kind help in critical reading of the manuscript. We also thank the State Government, Government of West Bengal for fellowship to RP.
Funding
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
Author information
Authors and Affiliations
Contributions
Rubia Parvin: Conceptualization, Investigation, Methodology, Formal analysis, Data curation, draw all the graphs,Writing-original draft. Khushnood Fatma: Methodology, Data curation. Debbithi Bera: Methodology, Data curation. Jyotirmayee Dash: Methodology, Data curation. Joydeep Mukherjhee: Conceptualization, Writing-review & editing, Validation, Formal analysis. Ratan Gachhui: Supervision, Conceptualization, Writing-review & editing, Validation, Resources, Formal analysis.
Corresponding author
Ethics declarations
Ethical approval
This article does not contain any experiments with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Parvin, R., Fatma, K., Bera, D. et al. The effect of temperature on the activity and stability of the thermostable enzyme caffeine dehydrogenase from Pichia manshurica CD1. Biologia 78, 3249–3258 (2023). https://doi.org/10.1007/s11756-023-01473-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11756-023-01473-9