Study on the extraction and purification of Chitin and Chitin based derivatives from Exoskeleton of Fresh water prawns
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Abstract
Crustacean’s shells constitute the traditional and current commercial source of Chitin. Chitin and its derivatives as a potential resource as well as multiple functional substrates have generated attractive interest in various fields such as biomedical, pharmaceutical, food and environmental industries. In the present investigation, chitinous wastes were collected from the fresh water areas of Dehradun and Rishikesh of Uttarakhand State. The bacterium, Bacillus sp. isolated from soil produces chitinase enzyme responsible for degradation of chitin obtained from chitinous wastes. Further the chitinases enzyme was utilized to degrade the chitinous wastes into chito-oligosaccharides. The chitin active molecule present in the chitinous waste at another stage was deacetylated to chitosan. Further the antibacterial activity of chitinases, chitin, chitosan and chito-oligosaccharides were determined in vitro by well diffusion method. The enzyme purified showed potent activity against the bacterial cultures, but no activity was observed against the fungal test cultures. Amongst the test bacterial cultures the chitinase showed maximum inhibition against Micrococcus luteus (diameter of zone of inhibition: 21 mm) followed by multi-drug resistant Staphylococcus aureus (diameter of zone of inhibition: 20 mm) and Salmonella abony (diameter of zone of inhibition: 17 mm). Further, chitin, chitosan and chito-oligosaccharide were subjected to antimicrobial activity against the similar strains and the results were found to be very satisfactory as the chitin and chitin-based derivatives were equally antimicrobial in nature. Further the antifungal activity of chitinases, chitin, chitosan and chito-oligosaccharides was determined in vitro by well diffusion method against Aspergillus niger and Candida albicans. Amongst the test fungal cultures, the chitinase showed maximum inhibition against Aspergillus niger (diameter of zone of inhibition: 24 mm) followed by Candida albicans (diameter of zone of inhibition: 14 mm). Deacetylated form of chitin i.e chitosan showed potent antifungal activity against Candida albicans (diameter of zone of inhibition: 24 mm) followed by Aspergillus niger (diameter of zone of inhibition: 18 mm). The chitin extracted showed almost similar antifungal activity against Aspergillus niger and Candida albicans (diameter of zone of inhibition: 15 mm) respectively. The low molecular weight derivatives viz. chito-oligosaccharide showed significant antifungal activity against Aspergillus niger (diameter of zone of inhibition: 14 mm) but no activity was found against Candida albicans.
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References
Aam BB, Heggset EB, Norberg AL, Sorlie M, Vaum KM, Eijsink VGH, Production of chitooligoosaccharides and their potential Applications in Medicine. Mar. Drugs, 2010; 8: 1482-1517.
Adachi, W.; Sakihama, Y.; Shimizu, S.; Sunami, T.; Fukazawa, T.; Suzuki, M.; Yatsunami, R.; Nakamura, S.; Takenaka, A. Crystal structure of family GH-8 chitosanase with subclass II specificity from Bacillus sp. K17. J. Mol. Biol. 2004, 343, 785–795.
Alam J., Kushwaha A., Mathur A. (2013). Evaluation of growth regulatory effect of Chitin and Chitin -based derivatives extracted from fresh water crustaceans. Recent Research in Science & Technology, 5 (1): 05-08.
Alam J., Mathur A. (2014). Antibacterial potential of Chitin and Chitin- based derivatives against pathogenic and drug resistant bacterial strains. World Journal of Pharmacy & Pharmaceutical Sciences, 3 (12): 1698-1707.
Alam J., Mathur A. (2014). Evaluation of antifungal potential of Chitin and Chitin-based derivatives against pathogenic fungal strains. Biolife, 2(4): 1354-1358.
Alam J., Mathur A. (2015). First Report on evaluation of anti-diabetic potential of Chitin and Chitin-based derivatives against alloxan induced albino rats. International Journal of Pharmaceutical Science Research, Accepted for publication in Vol. 6; Issue 9 (September, 2015); (Acknowledgement No. IJPSR/RA-5335/02-15)
Amatayakul-Chantler S, Ferguson, MAJ, Dwek RA, Rademacher TW, Parekh RB, Crandall, IE, Newell PC. Cell surface oligosaccharides on Dictyostelium during development. J. Cell Sci. 1975; 99: 485-495.
Andres, Y, Giraud, L.Gerente, C. & Le Cloirec, P. - Environ. Technol., 28, p.1357-1363 (2007).
Asaoka A. Chitin-chitosan –The choice food supplement for over 10, 000 physicians in Japan, New York, Vantage Press, Inc., 1996.
Assis, O. B. G, Bernardes Filho, R. Viera, D. C. & Campana Filho, S. P. - Int. J. Polymer. Mater., 51, p.633-638 (2002).
Bakkers J, Semino CE, Stroband H, Kijne JW, Robbins PW, Spaink HP. An important developmental role for oligosaccharides during early embryogenesis of cyprinid fish. Proc. Natl. Acad. Sci. USA. 1997; 94: 7982-7986.
Bautista-Banos S, Hernandez-Lopez M, Bosquez-Molina E, Wilson CL. Effects of chitosan and plant extracts on growth of Colletotrichum gloeosporioides, anthracnose levels and quality of papaya fruit. Crop.Prot. 2003; 22: 1087-1092.
Bhatnagar, A.; Sillanpää, M. Applications of chitin- and chitosan-derivatives for the detoxification of water and wastewater - A short review. Adv. Colloid Interface Sci. 2009, 152, 26–38.
Boot RG, Blommaart EFC, Swart E,ghauharali Van der Vlugt K, Bijl N, Moe C, Place A & Aerts JMG. Identification of the acidic mammalian chitinase distinct from Chitotriosidase. J Biol Chem. 2001 a; 276: 6770-6778.
Brameld, K.A.; Goddard, W.A. Substrate distortion to a boat conformation at subsite -1 is critical in the mechanism of family 18 chitinases. J. Am. Chem. Soc. 1998, 120, 3571–3580.
Brameld, K.A.; Goddard, W.A. The role of enzyme distortion in the single displacement mechanism of family 19 chitinases. Proc. Natl. Acad. Sci. USA 1998, 95, 4276–4281. Mar. Drugs 2010, 8 1509
Broekaert WF, Van Parijs J, Leyns F, Joos H, Peumans WJA. Chitin binding Lectin from stringing Nettle Rhizomes with Antifungal Properties. Sciences. 1989; 245: 1100-1102.
Buchsbaum, M. R. a. P, Vicki & John. (1987). Living Invertebrates. Pacific Grove, CA, The Boxwood Press.
Burkenroad, M. D. (1963). "The evolution of the Eucarida (Crustacea, Eumalacostraca), in relation to the fossil record". Tulane Studies in Geology 2 (1): 1–17.
Burkenroad, M. D. (1963). "The evolution of the Eucarida (Crustacea, Eumalacostraca), in relation to the fossil record". Tulane Studies in Geology 2 (1): 1–17.
Bussink AP, Speijer D, Aerts JM, Boot RG. Evolution of mammalian chitinase(-like) members of family 18 glycosyl hydrolases. Genetics. 2007; 177: 959-970.
Calabrese V, Lodi R, Tonon C, D'Agata V, Sapienza M, Scapagnini G, Mangiameli A, Pennisi G, Stella AMG, Butterfield DA. Oxidative stress, mitochondrial dysfunction and cellular stress response in Friedreich's ataxia. J. Neurol. Sci. 2005;233:145–162.
Cantarel, B.L.; Coutinho, P.M.; Rancurel, C.; Bernard, T.; Lombard, V.; Henrissat, B. The Carbohydrate-Active enZYmes database (CAZy): an expert resource for Glycogenomics. Nucleic Acids Res. 2009, 37, D233–D238.
Hirano S, Muzzarelli RAA, Peter MG. N-acyl,N-arylidene- and N-alkylidene chitosans and their hydrogels, Chitin handbook (European Chitin Society,Italy). 1997: pp.71-76.
Hoell IA, Dalmus B, Heggset EB, Aspmo SI & Eijsenki VGH. Crystal structure and enzymatic properties of a bacterial family 19 chitinase reveal differences from plant enzymes. FEBS. J. 2006; 273: 4889-4900.
Hon DN. Polysaccharides in Medicinal Applications; Marcel Dekker: New York. 1996; pp.631- 649.
Honee G. Engineered resistance against fungal plant pathogens. Eur J Plant pathol. 1999; 105: 319-326.
Horn, S.J.; Sørbotten, A.; Synstad, B.; Sikorski, P.; Sørlie, M.; Vårum, K.M.; Eijsink, V.G. Endo/exo mechanism and processivity of family 18 chitinases produced by Serratia marcescens. FEBS J. 2006, 273, 491–503.
Hudson S M & Jenkins D W, Chitin and Chitosan, Encyclopedia of polymer Science and Technology. Third ed(Wiley Intersciences,New York).
Hudson S M & Smith C. Polysaccharide: Chitin and chitosan:chemistry and technology of their use as structural materials, Biopolymers from renewable resources,edited by D L Kaplan, (Springer-Verlag, New York) 1988: pp.96-118.
Hurtado-Guerrero R, Dorfumeller HC, van Aalten DM. Molecular mechanisms of O-GlcNAcyclation. Curr.Opin. Struct. Biol. 2008; 18: 551-557.
Ikeyama H. RJ Morton eds. Chitin healing power from the sea, Los angeles, CA: Will productions,( 1995).
Imoto T and Yagishita K. A simple activity measurement by lysozyme. Agric Biol. Chem. 1971; 35:1154–1156.
Inui H, Tsujikobo M, Hirano S.(1995). Low molecular weight chitosan stimulation of mitogenic response to platelet deprived growth factor in vascular smooth muscle cells. Biosci., Biotech., Biochem.,59, 2111-2114.
Iseli, B.; Armand, S.; Boller, T.; Neuhaus, J.M.; Henrissat, B. Plant chitinases use two different hydrolytic mechanisms. FEBS Lett. 1996, 382, 186–188.
Ishihara M, Obara K, Nakamura S, Fujita M, Masuoka K, Kanatani Y, Takase B, Hattori H, Morimoto Y, Ishihara M, Maehara T, Kikuchi M. Chitosan hydrogel as a drug delievery carrier to control angiogenesis. J. Artif. Organs. 2006; 9: 8-16.
Itoh Y, Kawase T, Nikaidou N, Fukada H, Mitsutomi M, Watanabe T, Itoh Y. Functional analysis of the chitin binding domain of family 19 chitinase from streptomyces griseus HUT6307: Substrate-binding affinity and cis-dominant increase of antifungal function. Biosci. Biotechnol. Biochem. 2002; 66: 1084-1092.
J. K. Lowry (October 2, 1999). "Dendrobranchiata (Decapoda, Eucarida, Malacostraca)".Crustacea, the Higher Taxa. Australian Museum.
J. W. Martin & G. E. Davis (2001) (PDF). An Updated Classification of the Recent Crustacea. Natural History Museum of Los Angeles County. pp. 1–132.
J. W. Martin & G. E. Davis (2001) (PDF). An Updated Classification of the Recent Crustacea. Natural History Museum of Los Angeles County. pp. 1–132.
Janes K, Fresneau M, Marazuela A, Fabra A, Alonso M. Chitosan nanoparticles as delievery systems for doxorubicin. J. Controll. Release. 2001; 73: 255-267.
Jasrapuria S, Arakane Y, Osman G, Kramer KJ, Beeman RW, Muthukrishnan. Genes encoding proteins with peritrophin A-type chitin binding domains in Tribolium castaneum are grouped into three distinct families based on phulogeny, expression and function. Insect. Biochem. Mol. Biol. 2010; 40: 214-227.
Je J-Y, Kim S-K. Antioxidant activity of novel chitin derivative. Bioorg. Med. Chem. Lett. 2006;16:1884–1887. [PubMed]
Jeuniaux CA, Domard A, Jeuniaux C, Muzzarelli RAA, Roberts G, Eds Jacques, Andre Publ, Lyon France. Brief survey of the early contributionof European scientists to chitin knowledge. In Advances in Chitin Sciences. 1996: 1-9.
Mano JF, Silva GA, Azevedo HS, Malafaya PB, Sousa RA, Silva SS, Boesel LF, Oliveira JM, Santos TC, Marques AP, Neves NM, Reis RL. Natural origin biodegradable systems in tissue engineering and regenerative medicine: present status and some moving trends. J. R. Spoc. Interface. 2007; 4: 999-1030.
Marcotte, E.M.; Monzingo, A.F.; Ernst, S.R.; Brzezinski, R.; Robertus, J.D. X-ray structure of an anti-fungal chitosanase from Streptomyces N174. Nat. Struct. Biol. 1996, 3, 155–162.
Masson, M. Holappa, J. Hjalmarsdóttir, M. Runarsson, O. V. Nevalainen, T. & Jarvinen, T. - Carbohyd. Polym., 74, p.566-571 (2008).
Mathur A, Rawat A, Bhatt G, Baweja S, Ahmad F, Grover A, Madhav K, Dhand M, Mathur D, Verma SK, Singh SK, Dua VK. Isolation of Bacillus producing chitinase from soil: Production and purification of chito-oligosaccharides from chitin extracted from fresh water crustaceans and Antimicrobial activity of chitinase. Recent Res. Science & Technology.( 2011); 3(11): 01-06.