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Microbial Cholesterol Oxidase: Industrial Applications

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Microbial Enzymes: Roles and Applications in Industries

Part of the book series: Microorganisms for Sustainability ((MICRO,volume 11))

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Abstract

Cholesterol oxidase (ChOx) belongs to the oxidoreductase family and catalyzes the oxidation of cholesterol compounds. ChOx produced by bacteria, streptomyces, fungi, higher plants, and insects is being used in pharmaceutical formulations, agriculture, and waste management. Nowadays, application of ChOx in the detection of clinical diseases has also been reported. The industrial use and demand for ChOx have increased due to its effective and accurate functionality of bioconversion of cholesterol compounds into enantiomerically pure compounds used in various industries. Furthermore, ChOx has also emerged as an insecticide and showed great potential in replacing conventional chemical agents used in the control of cotton pest. In this chapter, we accentuate on the structure, sources, production, and commercial applications of ChOx.

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References

  • Allain CC, Poon LS, Chan CS, Richmond W, Fu PC (1974) Enzymatic determination of total serum cholesterol. Clin Chem 20:470–475

    Article  CAS  PubMed  Google Scholar 

  • Aparicio JF, Martin JF (2008) Microbial cholesterol oxidases: bioconversion enzymes or signal proteins. Mol BioSyst 4:804–809

    Article  CAS  PubMed  Google Scholar 

  • Arya SK, Datta M, Malhotra BD (2008) Recent advances in cholesterol biosensor. Biosens Bioelectron 23:1083–1100

    Article  CAS  PubMed  Google Scholar 

  • Atrat PG, Wagner B, Wagner M, Schumann G (1992) Localization of the cholesterol oxidase in Rhodococcus erythropolis IMET 7185 studied by immunoelectron microscopy. J Steroid Biochem Mol Bio 42:193–200

    Article  CAS  Google Scholar 

  • Binod P, Sindhu R, Pandey A (2013) The alcohol fermentation step: The most common ethanologenic microorganisms among yeasts, bacteria and filamentous fungi. In: Faraco V (ed) Lignocellulose conversion: Enzymatic and microbial tools for bioethanol production. Springer, Berlin, pp 131–150

    Chapter  Google Scholar 

  • Bongiovanni C, Ferri T, Poscia A, Varalli M, Santucci R, Desideri A (2001) An electrochemical multienzymatic biosensor for determination of cholesterol. Bioelectrochemistry 54:17–22

    Article  CAS  PubMed  Google Scholar 

  • Bru R, Sánchez FA, García CF (1989) Characterization of cholesterol oxidase activity in AOT-isooctane reverse micelles and its dependence on micelle size. Biotechnol Lett 11:237–242

    Article  CAS  Google Scholar 

  • Brzostek A, Dziadek B, Rumijowska GA, Pawelczyk J, Dziadek J (2007) Cholesterol oxidase is required for virulence of Mycobacterium tuberculosis. FEMS Microbiol Lett 275:106–112

    Article  PubMed  CAS  Google Scholar 

  • Caldinelli L, Iametti S, Barbiroli A, Bonomi F, Fessas D, Molla G, Pilone MS, Pollegioni L (2005) Dissecting the structural determinants of the stability of cholesterol oxidase containing covalently bound flavin. J Biol Chem 280:22572–22581

    Article  CAS  PubMed  Google Scholar 

  • Cheetham PSJ, Dunnill P, Lilly MD (1982) The characterization and interconversion of three forms of cholesterol oxidase extracted from Nocardia rhodochrous. Biochem J 201:512–521

    Article  Google Scholar 

  • Chen L, Lyubimov AY, Brammer L, Vrielink A, Sampson NS (2007) The binding and release of oxygen and hydrogen peroxide are directed through a hydrophobic tunnel in cholesterol oxidase. Biochemistry 47:5368–5377

    Article  CAS  Google Scholar 

  • Coulombe R, Yue KQ, Ghisla S, Vrielink A (2001) Oxygen access to the active site of cholesterol oxidase through a narrow channel is gated by an Arg-Glu pair. J Biol Chem 276:30435–30441

    Article  CAS  PubMed  Google Scholar 

  • Croteau N, Vrielink A (1996) Crystallization and preliminary X-ray analysis of cholesterol oxidase from Brevibacterium sterolicum containing covalently bound FAD. J Struct Biol 116:317–319

    Article  CAS  PubMed  Google Scholar 

  • Dewan SS (2011) Global markets for enzymes in industrial applications (BIO030G). BCC Research

    Google Scholar 

  • Dewan SS (2012) Global Markets and Technologies for Biofuel Enzymes (EGY099A). BCC Research

    Google Scholar 

  • Doukyu N (2009) Characteristics, biotechnological applications of microbial cholesterol oxidases. Appl Microbiol Biotechnol 83:825–837

    Article  CAS  PubMed  Google Scholar 

  • Doukyu N, Aono R (2001) Cloning, sequence analysis and expression of a gene encoding and organic solvent and detergent-tolerant cholesterol oxidase of Burkholderia cepacia strain ST-200. Appl Microbiol Biotechnol 57:146–152

    Article  CAS  PubMed  Google Scholar 

  • Doukyu N, Nihei S (2015) Cholesterol oxidase with high catalytic activity from Pseudomonas aeruginosa: screening, molecular genetic analysis, expression and characterization. J Biosci Bioeng 120(1):24–30

    Article  CAS  PubMed  Google Scholar 

  • Doukyu N, Shibata K, Ogino H, Sagermann M (2008) Purification and characterization of Chromobacterium sp. DS-1 cholesterol oxidase with thermal, organic solvent and detergent tolerance. Appl Microbiol Biotechnol 80:59–70

    Article  CAS  PubMed  Google Scholar 

  • Edwards DP, O’Conner JL, Bransome ED, Braselton WE (1976) Human placental 3b-hydroxysteroid dehydrogenase: D5-Isomerase. J Biol Chem 251:1632–1638

    CAS  PubMed  Google Scholar 

  • El-Naggar NE, Deraz SF, Soliman HM, El-Deeb NM, El-Shweihy NM (2017) Purification, characterization and amino acid content of cholesterol oxidase produced by Streptomyces aegyptia NEAE 102. BMC Microbiol 17(1):76

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ernst ND, Cleeman JI (2002) National cholesterol education program keeps a priority on lifestyle modification to decrease cardiovascular disease risk. Curr Opin Lipidol 13:69–73

    Article  CAS  PubMed  Google Scholar 

  • Espenshade PJ, Hughes AL (2007) Regulation of sterol synthesis in eukaryotes. Annu Rev Genet 41:401–427

    Article  CAS  PubMed  Google Scholar 

  • Eventoff W, Rossmann MG (1975) The evolution of dehydrogenases and kinases. CRC Crit Rev Biochem 3:111–140

    Article  CAS  PubMed  Google Scholar 

  • Fazaeli A, Golestani A, Lakzaei M, Rasi Varaei SS, Aminian M (2019) Expression optimization, purification, and functional characterization of cholesterol oxidase from Chromobacterium sp. DS1. PLoS One 14:e0212217

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fukuyama M, Miyake Y (1979) Purification and some properties of cholesterol oxidase from Schizophyllum commune with covalently bound flavin. J Biochem 85:1183–1193

    CAS  PubMed  Google Scholar 

  • Ghoshroy KB, Zhu W, Sampson NS (1997) Investigation of membrane disruption in the reaction catalyzed by cholesterol oxidase. Biochemistry 36(20):6133–6140. https://doi.org/10.1021/bi962190p

    Article  CAS  PubMed  Google Scholar 

  • Giorgi V, Menéndez P, García-Carnelli C (2019) Microbial transformation of cholesterol: reactions and practical aspects—an update. World J Microbiol Biotechnol 35(9):131

    Article  PubMed  CAS  Google Scholar 

  • Greenplate JT, Duck NB, Pershing JC, Purcell JP (1995a) Cholesterol oxidase: an oostatic and larvicidal agent active against the cotton boll weevil Anthonomus grandis. Entomol Exp Appl 74:253–258

    Article  CAS  Google Scholar 

  • Greenplate JT, Duck NB, Pershing JC, Purcell JP (1995b) Cholesterol oxidase: an oostatic and larvicidal agent active against the cotton boll weevil, Anthonomus grandis. Entomol Exp Appl 74:253–258

    Article  CAS  Google Scholar 

  • Guo FQ, Okamoto M, Crawford NM (2003) Identification of a plant nitric oxide synthase gene involved in hormonal signaling. Science 302:100–103

    Article  CAS  PubMed  Google Scholar 

  • Hao M, Fan G, Zhang Y, Xin Y, Zhang L (2019) Preparation and characterization of copper-Brevibacterium cholesterol oxidase hybrid nanoflowers. Int J Biol Macromol 126:539–548

    Article  CAS  PubMed  Google Scholar 

  • Hol WG, Duijnen VPT, Berendsen HJ (1978) The alpha-helix dipole and the properties of proteins. Nature 273:443–446

    Article  CAS  PubMed  Google Scholar 

  • Illanes A (2008) Enzyme production. In: Illanes A (ed) Enzyme biocatalysis: principles and applications. Springer Netherlands, Dordrecht, pp 57–106

    Chapter  Google Scholar 

  • Isobe K, Mori N, Wakao N (2003a) The second cholesterol oxidase produced by gamma-proteobacterium Y-134. J Biosci Bioeng 96(3):257–261

    Article  CAS  PubMed  Google Scholar 

  • Isobe K, Shoji K, Nakanishi Y, Yokoe M, Wakao N (2003b) Purification and some properties of cholesterol oxidase stable in detergents from gamma- Proteobacterium Y-134. J Biosci Bioeng 95:257–263

    Article  CAS  PubMed  Google Scholar 

  • Kim K, Rhee C, Park H (2002) Degradation of cholesterol by Bacillus subtilis SFF34 isolated from Korean traditional fermented flatfish. Lett Appl Microbiol 35:468–472

    Article  CAS  PubMed  Google Scholar 

  • Kreit J, Lefebvre G, Germain P (1994) Membrane-bound cholesterol oxidase from Rhodococcus sp. cells: production and extraction. J Biotechnol 33:271–282

    Article  CAS  Google Scholar 

  • Lartillot S, Kedziora P (1990) Production, purification and some properties of cholesterol oxidase from a Streptomyces sp. Prep Biochem 20:51–62

    CAS  PubMed  Google Scholar 

  • Lashkarian H, Raheb J, Shahzamani K, Shahbani H, Shamsara M (2010) Extracellular cholesterol oxidase from Rhodococcus sp.: isolation and molecular characterization. Iran Biomed J 14(1–2):49–57

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lee SY, Rhee HI, Tae WC, Shin JC, Park BK (1989) Purification and characterization of cholesterol oxidase from Pseudomonas sp. and taxonomic study of the strain. Appl Microbiol Biotechnol 31:542–546

    Article  CAS  Google Scholar 

  • Li S, Yang X, Yang S, Zhu M, Wang X (2012) Technology prospecting on enzymes: application, marketing and engineering. Comput Struct Biotechnol J 2:e201209017. https://doi.org/10.5936/csbj.201209017

    Article  PubMed  PubMed Central  Google Scholar 

  • Lin Y, Fu J, Song X (2012) Purification and characterization of an extracellular cholesterol oxidase from a Bordetella sp. Process Biochem 45:1563–1569

    Article  CAS  Google Scholar 

  • Liu WH, Meng MH, Chen KS (1988) Purification and some properties of cholesterol oxidases produced by an inducible and a constitutive mutant of Arthrobacter simplex (microbiology and fermentation industry). Agric Biol Chem 52:413–418

    Article  CAS  Google Scholar 

  • MacLachlan J, Wotherspoon AT, Ansell RO, Brooks CJ (2000) Cholesterol oxidase: sources, physical properties and analytical applications. J Steroid Biochem Mol Biol 72:169–195

    Article  CAS  PubMed  Google Scholar 

  • Matsui I, Nakashima K, Taniguchi T et al. (1982) Preparation of cholesterol oxidase. Patent No JP5757122792

  • Mendes MV, Recio E, Anton N, Guerra SM, Santos AJ, Martin JF, Aparicio JF (2007) Cholesterol oxidases act as signaling proteins for the biosynthesis of the polyene macrolide pimaricin. Chem Biol 14:279–290

    Article  CAS  PubMed  Google Scholar 

  • Navas J, Gonzalez-Zorn B, Ladron N, Garrido P, Vazquez-Boland JA (2001) Identification and mutagenesis by allelic exchange of choE, encoding a cholesterol oxidase from the intracellular pathogen Rhodococcus equi. J Bacteriol 183:4796–4805

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Norouzi P, Faridbod F, Nasli Esfahani E, Larijani B, Ganjali M (2010) Cholesterol biosensor based on MWCNTs-MnO2 nanoparticles using FFT continuous cyclic voltammetry. Int J Electrochem Sci 5:1008–1017

    CAS  Google Scholar 

  • Ohlsson I, Nordstrom B, Branden CI (1974) Structural and functional similarities within the coenzyme binding domains of dehydrogenases. J Mol Biol 89:339–354

    Article  CAS  PubMed  Google Scholar 

  • Ohta T, Fujishiro K, Yamaguchi K, Tamura Y, Aisaka K, Uwajima T, Hasegawa M (1991) Sequence of gene choB encoding cholesterol oxidase of Brevibacterium sterolicum: comparison with choA of Streptomyces sp. SA-COO. Gene 103:93–96

    Article  CAS  PubMed  Google Scholar 

  • Pei Y, Dupont C, Sydor T, Haas A, Prescott JF (2006) Cholesterol oxidase (ChoE) is not important in the virulence of Rhodococcus equi. Vet Microbiol 118:240–246

    Article  CAS  PubMed  Google Scholar 

  • Pollegioni L, Piubelli L, Molla G (2009) Cholesterol oxidase: biotechnological applications. FEBS J 276:6857–6870

    Article  CAS  PubMed  Google Scholar 

  • Prakash D, Nawani N, Prakash M, Bodas M, Mandal A, Khetmalas M, Kapadnis B (2013) Actinomycetes: a repertory of green catalysts with a potential. Biomed Res Int 2013:264020, 1-8

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Praveen V, Srivastava A, Tripathi C (2011) Purification and characterization of the enzyme cholesterol oxidase from a new isolate of Streptomyces sp. Appl Biochem Biotechnol 165:1414–1426

    Article  CAS  PubMed  Google Scholar 

  • Purcell JP, Greenplate JT, Jennings MG, Ryerse JS, Pershing JC, Sims SR, Prinsen MJ, Corbin D, Tran RM, Sammons RD (1993a) Cholesterol oxidase: a potent insecticidal protein active against boll weevil larvae. Biochem Biophys Res Commun 196:1406–1413

    Article  CAS  PubMed  Google Scholar 

  • Purcell JP, Greenplate JT, Jennings MG, Ryerse JS, Pershing JC, Sims SR, Prinsen MJ, Corbin DR, Tran M, Sammons RD, Stonard RJ (1993b) Cholesterol oxidase: a potent insecticidal protein active against boll weevil larvae. Biochem Biophys Res Commun 196(3):1406–1413

    Article  CAS  PubMed  Google Scholar 

  • Purcell JP, Greenplate JT, Jennings MG, Ryerse JS, Pershing JC, Sims SR et al (1993c) Cholesterol oxidase: a potent insecticidal protein active against boll weevil larvae. Biochem Biophys Res Commun 196(3):1406–1413

    Article  CAS  PubMed  Google Scholar 

  • Qiaocui S, Tuzhi P, Yunu Z, Yang CF (2005) An electrochemical biosensor with cholesterol oxidase sol-gel film on a nanoplatinum/carbon nanotube electrode. Electroanalysis 17:857–861

    Article  CAS  Google Scholar 

  • Randolph T, Clark D, Blanch H, Prausnitz J (1988) Enzymatic oxidation of cholesterol aggregates in supercritical carbon dioxide. Science 239:387–390

    Article  CAS  PubMed  Google Scholar 

  • Reiss R, Faccio G, Thöny-Meyer L, Richter M (2014) Cloning, expression and biochemical characterization of the cholesterol oxidase CgChoA from Chryseobacterium gleum. BMC Biotechnol 14:46

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Richmond W (1973) Preparation and properties of a cholesterol oxidase from Nocardia sp. and its application to the enzymatic assay of total cholesterol in serum. Clin Chem 19:1350–1356

    Article  CAS  PubMed  Google Scholar 

  • Sahu S, Shera SS, Banik RM (2019) Optimization of process parameters for cholesterol oxidase production by Streptomyces olivaceus MTCC 6820. Open Biotechnol J 13(1)

    Google Scholar 

  • Salva TJ, Liserre AM, Moretto AL, Zullo MA, Ventrucci G, Menezes TJ (1999) Some enzymatic properties of cholesterol oxidase produced by Brevibacterium sp. Rev Microbiol 30(4):315–323

    Article  CAS  Google Scholar 

  • Sampson NS, Vrielink A (2003) Cholesterol oxidases: a study of nature's approach to protein design. Acc Chem Res 36:713–722

    Article  CAS  PubMed  Google Scholar 

  • Schatz A, Savard K, Pintner IJ (1949) The ability of soil microorganisms to decompose steroids. J Bacteriol 58:117–125

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shi QC, Zhi TP (2005) A novel cholesterol oxidase biosensor based on pt-nanoparticle/carbon nanotube modified electrode. Chin Chem Lett 16(8):1081–1084

    CAS  Google Scholar 

  • Shirokane YNK, Mizusawa K (1977) Purification and some properties of an extracellular 3-hydroxysteroid oxidase produced by Corynebacterium cholesterolicum. J Ferment Tech 55:337–345

    CAS  Google Scholar 

  • Smith AG, Brooks CJW (1976) Cholesterol oxidases: properties and applications. J Steroid Biochem 7:705–713

    Article  CAS  PubMed  Google Scholar 

  • Smith M, Zahnley J, Pfeifer D, Goff D (1993) Growth and cholesterol oxidation by Mycobacterium species in Tween 80 medium. Appl Environ Microbiol 59:1425–1429

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sojo M, Bru R, Lopez-Molina D, Garcia-Carmona F, Arguelles JC (1997) Cell linked and extracellular cholesterol oxidase activities from Rhodococcus erythropolis. Isolation and physiological characterization. Appl Microbiol Biotechnol 47:583–589

    Article  CAS  PubMed  Google Scholar 

  • Srivastava A, Singh V, Shafiul H, Pandey S, Mishra M et al (2018a) Response surface methodology-genetic algorithm based medium optimization, purification, and characterization of cholesterol oxidase from Streptomyces rimosus. Sci Rep 8:10913

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Srivastava A, Singh V, Tripathi CKM (2018b) Scale up and optimization of cholesterol oxidase production from Streptomyces rimosus MTCC10792 in a 3-L bioreactor. Environ Sustain 1(1)

    Google Scholar 

  • Stepankova V, Bidmanova S, Koudelakova T, Prokop Z, Chaloupkova R, Damborsky J (2013) Strategies for stabilization of enzymes in organic solvents. ACS Catal 3(12):2823–2836

    Article  CAS  Google Scholar 

  • Turfitt GE (1944) The microbiological degradation of steroids: 2. Oxidation of cholesterol by Proactinomyces sp. Biochem J 38:492–496

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Varma R, Nene S (2003) Biosynthesis of cholesterol oxidase by Streptomyces lavendulae NCIM 2421. Enzym Microb Technol 33:286–291

    Article  CAS  Google Scholar 

  • Vidal JC, Espuelas J, Castillo JR (2004) Amperometric cholesterol biosensor based on in situ reconstituted cholesterol oxidase on an immobilized monolayer of flavin adenine dinucleotide cofactor. Anal Biochem 333:88–98

    Article  CAS  PubMed  Google Scholar 

  • Vrielink A, Ghisla S (2009) Cholesterol oxidase: biochemical properties. FEBS J 276:6826–6843

    Article  CAS  PubMed  Google Scholar 

  • Watanabe K, Aihara H, Nakamura R, Suzuki KI, Momagata K (1986) Isolation and identification of cholesterol degradation Rhodococcus strains food of animal origin and their cholesterol oxidase activities. J Gen Appl Microbiol 32:137–147

    Article  CAS  Google Scholar 

  • Wilmanska D, Sedlaczek L (1988) The kinetics of biosynthesis and some properties of an extracellular cholesterol oxidase produced by Arthrobacter sp. IM 79. Acta Microbiol Pol 37:45–51

    CAS  Google Scholar 

  • Yamada K, Koroleva A, Laughlin M, Oksanen N, Akhgari A, Safronova V, Yakovleva E, Kolodyaznaya V, Buldakova T, Metsä-Ketelä M (2019) Characterization and overproduction of cell-associated cholesterol oxidase ChoD from Streptomyces lavendulae YAKB-15. Sci Rep 9(1):11850

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Yazdi MT, Malekzadeh F, Zarrini G, Faramarzi MA, Kamranpour N, Khaleghparast S (2001) Production of cholesterol oxidase by a newly isolated Rhodococcus sp. World J Microbiol Biotechnol 17:731–737

    Article  CAS  Google Scholar 

  • Ye D, Lei J, Li W, Ge F, Wu K, Xu W, Yong B (2008) Purification and characterization of extracellular cholesterol oxidase from Enterobacter sp. World J Microbiol Biotechnol 24:2227–2233

    Article  CAS  Google Scholar 

  • YıldırımoÄŸlu F, Arslan F, Çete S, YaÅŸar A (2009) Preparation of a polypyrrole polyvinyl sulphonate composite film biosensor for determination of cholesterol based on entrapment of cholesterol oxidase. Sensors (Basel) 9:6435–6445

    Article  CAS  Google Scholar 

  • Yue QK, Kass IJ, Sampson NS, Vrielink A (1999) Crystal structure determination of cholesterol oxidase from Streptomyces and structural characterization of key active site mutants. Biochemistry 38:4277–4286

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We are thankful to Naveen Kumar Arora and Jitendra Mishra for providing editorial contribution. We are also gratified to Jitendra Mishra for preparing color illustration. The authors thank Dr. Qazi Mohd. Sajid Jamal (Department of Heath Informatics, College of Public Health Informatics, Qassim University, Saudi Arabia) for providing the assistance in computational modelling data.

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Akanksha, Mishra, V., Kesari, K.K. (2020). Microbial Cholesterol Oxidase: Industrial Applications. In: Arora, N., Mishra, J., Mishra, V. (eds) Microbial Enzymes: Roles and Applications in Industries. Microorganisms for Sustainability, vol 11. Springer, Singapore. https://doi.org/10.1007/978-981-15-1710-5_12

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