Study on the Effect of Blending Ratios on the Antibacterial Activities of Chitosan/Gelatin Blend Solutions

혼합비율에 따른 키토산/젤라틴 혼합용액의 항균활성에 관한 연구

  • Published : 2005.06.30

Abstract

Chitosan, second largest biomass after cellulose on earth, has potential for use as functional food package due to its antibacterial activity. However, due to high melting temperature of chitosan, chitosan films have been made by casting method. Because gelatin has relatively low molting temperature depending upon amount of plasticizer added, it was added to chitosan to produce commercially feasible film. The objective of the current study was to determine optimum blend ratio and amount of chitosan/gelatin blend solutions against antibacterial activities for extruder resin. Gram-positive bacteria (Bacillus cereus ATCC 14579 and Listeria monocytogenes ATCC 15313) and -negative bacteria (Escherichia coli ATCC 25922 and Salmonella enteritidis IFO 3313) were used. Paper (8 mm) diffusion and optical density methods were used to evaluate effect of different blending ratio solutions on the inhibition of bacterial growth. Measured clear none size ranged from 8 mm to 18.07 mm in paper diffusion test. For B. cereus, E. coli, and S. enteritidis, addition of $50\;{\mu}L$ blend solution (chitosan/gelatin = 2/8: 0.3 mg) resulted in clear zone on paper disc. In L. monocytogenes, inhibition effect was observed with 0.6 mg chitosan (chitosan/gelatin=4/6). Minimum inhibitory concentration (MIC) values of B. cerues, L. monocytogenes, E. coli, and S. enteritidis with addition of chitosan were 0.1461, 0.2419, 0.0980, and 0.0490 mg/mL, respectively, These results indicate possibility of producing commercially feasible film with addition of optimum chitosan/gelatin amount.

Keywords

biopolymer;chitosan;gelatin;blending ratios;antibacterial activity

References

  1. Jo HL. Antimicrobial activity and food preservative function of a low molecular weight chitosan. Ph. D. thesis. Pusan National Fisheries Univ., Pusan, Korea (1989)
  2. Shahidi F, Arachchi JKV, You JJ. Food applications of chitin and chitosan. Trends Food Sci. Technol. 10: 37-51 (1999) https://doi.org/10.1016/S0924-2244(99)00017-5
  3. SAS Institute, Inc. SAS User's Guide. Statistical Analysis Systems Institute, Cary, NC, USA (1990)
  4. Hwang JK, Kim HJ, Shim JS, Pyun YP. Bacteriocidal activity of chitosan on Streptococcus mutans. Korean J. Food Sci. Technol. 31:522-526(1999)
  5. Sudarshan NR, Hoover DG Knorr D. Antibacterial action of chitosan. Food Biotechnol. 6: 257-272 (1992) https://doi.org/10.1080/08905439209549838
  6. Oh SW, Hong SP, Kim HJ, Choi YJ. Antimicrobial effects of chitosans on Escherichia coli O157:H7, Staphyloccus aureus and Candida albicans. Korean J. Food Sci. Technol. 32: 218-224 (2000)
  7. Kendra DF, Hadwiger LA. Characterization of the smallest chitosan oligomer that is maximally antifungal to Fusarium solani and elicits pisatin formation in Pisum sativum. Exp. Mycol. 8: 276-281 (1984) https://doi.org/10.1016/0147-5975(84)90013-6
  8. Tokura S, Ueno K, Miyazaki S, Nishi N. Molecular weight dependent antimicrobial activity by chitosan. Macromolecular Symposia 120: 1-9(1997) https://doi.org/10.1002/masy.19971200103
  9. Uchida Y, Izume M, Ohtakara A. Purification and Enzymatic properties of Chitosanase from Bacillus sp. M. Bull. Fac. Agr. Saga Univ. 66: 105-116(1989)
  10. Koyano T, Minoura N, Nagura M, Kobayashi K. Attachment and growth of cultured fibrablast cells on PVA/chitosan blended hydrogels. Biomed. Mater. Res. 39:486-490 (1998) https://doi.org/10.1002/(SICI)1097-4636(19980305)39:3<486::AID-JBM20>3.0.CO;2-7
  11. Arai K, Kinumaki T, Fugita T. Toxicity of chitosan. Bull. Tokai Reg. Fich Res. Lab. 56: 86-94 (1968)
  12. Zhao L, Mitomo H, Zhai M, Yoshii F, Nagasawa N, Kume T. Synthesis of antibacterial PVA/CM-chitosan blend hydrogels with electron beam irradiation. Carbohydr. Polymers, 53: 439-446 (2003) https://doi.org/10.1016/S0144-8617(03)00103-6
  13. Muzzarelli RAA, Barontini G, Rocchetti R. Immobilized enzymes on chitosan columns: $\alpha$ chymotrypsin and acid phosphatase. Biotechnol. Bioeng. 18: 1445 (1976)
  14. Li Q, Dunn T, Grandmaison EW, Goosen MFA. Applications and properties of chitosan. pp. 3-30. In: Applications of chitin and chitosan, Goosen MFA (ed). Technomic Publishing Co., Lancaster, UK (1997)
  15. Berkely RCW, Gooday GW, Ellwood DC. Chitin chitosan and their degradative enzymes, pp. 204-250. In Microbial Polysaccharides and Polysaccharides. Berkely RCW (ed). Academic Press, New York, NY, USA (1979)
  16. Gilsenan PM, Ross-Murphy SB. Rheology characterization of gelatins from mammalian and marine sources. Food Hydrocolloids 14: 191-195 (2000) https://doi.org/10.1016/S0268-005X(99)00050-8
  17. Allan CR, Hadwiger LA. The fungicidal effects of chitosan on fungi and varying cell wall composition. Exp. Mycol. 3: 285-287 (1979) https://doi.org/10.1016/S0147-5975(79)80054-7
  18. Kim MH, Oh SW, Hong SP, Yoon SK. Antimicrobial characteristics of chitosan and chitosan oligosaccharides on the microorganisms related to kimchi. Korean J. Food Sci. Technol. 30: 1439-1447(1998)
  19. Chung YC, Wang HL, Chen YM, Li SL. Effect of abiotic factors on the antibacterial activity of chitosan against waterborne pathogens. Bioresource Technol. 88: 179-184 (2003) https://doi.org/10.1016/S0960-8524(03)00002-6
  20. Liu XF, Guan YL, Yang DZ, Yao KD. Antibacterial action of chitosan and carboxymethylated chitosan. J. Appl. Polym. Sci. 79:1324-1335(2001) https://doi.org/10.1002/1097-4628(20010214)79:7<1324::AID-APP210>3.0.CO;2-L
  21. Cha DS, Cooksey K, Chinnan MS, Park HJ. Release of nisin from various heat-pressed and cast films. Lebensm.-Wiss. U.-Technol. 36: 209-213 (2003) https://doi.org/10.1016/S0023-6438(02)00209-8
  22. Bigi A, Bracci B, Cojazzi G, Panzavolta S, Roveri N. Drawn gelatin films with improved mechanical properties. Biomaterials 19: 2335-2340(1998) https://doi.org/10.1016/S0142-9612(98)00149-5
  23. Ross. Murphy SB. Structure and rheology of gelatin gels. Polymer 33: 2622-2627 (1992) https://doi.org/10.1016/0032-3861(92)91146-S
  24. Sobral PJA, Menegalli FC, Hubinger MD, Roques MA. Mechanical, water vapor barrier and thermal properties of gelatin based edible films. Food Hydrocolloids 15: 423-432 (2001) https://doi.org/10.1016/S0268-005X(01)00061-3
  25. Yamaguchi H. Application of chitin/chitosan to food and medicine fields. Shokuhin to Kaihatsu. 21: 203 (1986)
  26. Conner DE, Beuchat LR. Effect of essential oil from plants on growth of food spoilage yeast. J. Food Sci. 49: 429-434 (1984) https://doi.org/10.1111/j.1365-2621.1984.tb12437.x
  27. Toroko A, Tatewaki N, Suzuki K, Mikami T, Suzuki S, Suzuki M. Growth inhibitory effect of hexa-N-acetylchitohexaose against Meth-A solid turner. Chem. Pharm. Bull. 36: 784-790 (1988) https://doi.org/10.1248/cpb.36.784
  28. Stossel P, Leuba JL. Effect of chitosan, chitin and some amino-sugars on growth of various soilborns phytopathogenic fungi. Phytopath. W. 111: 82-90 (1984) https://doi.org/10.1111/j.1439-0434.1984.tb04244.x
  29. Normand V, Muller S, Ravey JC, Parker A. Gelation kinetics of gelation: a master curve and network modeling. Macromolecules. 33: 1063-1071 (2000) https://doi.org/10.1021/ma9909455
  30. Choi YH, Lim ST, Yoo BS. Dynamic rheological properties of gelatin. Korean J. Food Sci. Technol. 34: 830-834 (2002)
  31. Wu J, Chiu SC, Rearce EM, Kwei TK. Effects of phenolic compounds on gelation behavior of gelatin gels. J. Polym. Sci. Part A: Polym. Chem. 39: 224-231 (2000) https://doi.org/10.1002/1099-0518(20010101)39:1<224::AID-POLA250>3.0.CO;2-X
  32. Bigi A, Panzavolta S, Roveri N. Hydroxyapatite-gelatin films: a structural and mechanical characterization. Biomaterials 19: 739-744(1998) https://doi.org/10.1016/S0142-9612(97)00194-4