PRODUCTION AND CHARACTERIZATION OF CHITOOLIGOSACCHARIDE HYDROLYSATE PREPARED FROM CHITOSANASE ENZYME OF MARINE ISOLATES

Authors

  • P. VANATHI Department of Microbiology, Bharathidasan Arts and Science College, Ellispettai, Erode, Tamil Nadu, India https://orcid.org/0009-0001-5899-5501

DOI:

https://doi.org/10.22159/ijpps.2024v16i8.51671

Keywords:

COS, Marine origin, Diabetic foot ulcer, Fourier transform infrared spectroscopy (FT-IR), Nuclear magnetic resonance (NMR), X-ray diffraction (XRD) analysis, Antimicrobial activity

Abstract

Objective:  The present study was carried out to develop an enzymatic hydrolysate with unique biological properties targeting diabetic foot ulcers.

Methods: Chitosanase-producing organisms were isolated and used to create chitooligosaccharide hydrolysate. Various techniques, such as FTIR, NMR, and X-ray diffraction, were used. Antimicrobial activity was tested using disc diffusion and well diffusion methods. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined through the Chitooligosaccharide-Broth Dilution Method.

Results: The study identified marine mud samples and isolated S9, S15, and SF12 as significant sources of chitosanase production. The partially purified chitosanolytic enzymes produced by these isolates were hydrolyzed in a 1% chitosan solution at 180 °C, revealing more prominent antimicrobial activity. The Chitooligosaccharide Hydrolysate (COS) preparation was fixed at 45 °C, pH 5.5, for 180 minutes. The chitosanase enzyme was soluble in four solvents and insoluble in ethanol, acetone, and diethyl ether. All COS hydrolysates prepared showed antimicrobial activity against foot ulcer pathogens, Pseudomonas sp., and Candida albicans. S9 COS showed higher activity than SF12 hydrolysates against foot ulcer pathogens. The COS hydrolysate showed significantly stronger antimicrobial activities than chitosan and chitosanase.

Conclusion: The present study concludes that COS hydrolysate and its biological functions are applicable for diabetic foot ulcer treatment. Further investigation into the efficacy of COS against diverse infectious pathogens is needed.

Downloads

Download data is not yet available.

References

Xue T, Wang W, Yang Z, Wang F, Yang L, Li J, Gan H, Gu R, Wu Z, Dou G, Meng Z. Accurate determination of the degree of deacetylation of chitosan using USsceriaPLC–MS/MS. Int J Mole Sci. 2022 Aug 8;23(15):8810.

Fang Z, Cong W, Zhou H, Zhang J, Wang M. Biological activities, mechanisms and applications of chitooligosaccharides in the food industry. J Funct Foods. 2024 May 1;116:106219.

Jiménez-Gómez CP, Cecilia JA. Chitosan: a natural biopolymer with a wide and varied range of applications. Molecules. 2020 Sep 1; 25(17):3981.

Bonin M, Sreekumar S, Cord-Landwehr S, Moerschbacher BM. Preparation of defined chitosan oligosaccharides using chitin deacetylases. Int J Mol Sci. 2020 Qct 22; 21(21):7835. Published 2020 Oct 22

Huq T, Khan A, Brown D, Dhayagude N, He Z, Ni Y. Sources, production and commercial applications of fungal chitosan: A review. J Bioresour Bioprod. 2022 May 1;7(2):85-98.

Abdelhamid HN. Chitosan-based nanocarriers for gene delivery. Nanoengineering of Biomaterials. 2022 Feb 14; 91-105.

Bai L, Liu L, Esquivel M, et al. Nanochitin: chemistry, structure, assembly, and applications. Chem Rev. 2022 Jun 2; 122(13):11604-11674.

Ssekatawa K, Byarugaba DK, Wampande EM, et al. Isolation and characterization of chitosan from Ugandan edible mushrooms, Nile perch scales and banana weevils for biomedical applications. Sci Rep. 2021 Feb 18; 11(1):4116.

Khan F, Pham DT, Oloketuyi SF, Manivasagan P, Oh J, Kim YM. Chitosan and their derivatives: antibiofilm drugs against pathogenic bacteria. Colloids Surf B Biointerfaces. 2020 Jan 1; 185:110627.

Boi VN, Trang NT, Cuong DX, Hoan VT, Hai L. Oligosaccharide Chitosan: Viscosity, Molecular Weight, Antibacterial Activity, and Impact of γ Radiation. World. 2020 Apr 29 ;4(2):40-5.

Ul-Islam M, Alabbosh KF, Manan S, Khan S, Ahmad F, Ullah MW. Chitosan-based nanostructured biomaterials: synthesis, properties, and biomedical applications. Adv Ind Eng Polym Res. 2024 Jaan 1; 7:79-99.

Salama A, Hasanin M, Hesemann P. Synthesis and antimicrobial properties of new chitosan derivatives containing guanidinium groups. Carbohydr Polym. 2020 Aug 1; 241:116363.

Ke CL, Deng FS, Chuang CY, Lin CH. Antimicrobial actions and applications of chitosan. Polymers. 2021 Mar 15; 13(6):904.

Desai N, Rana D, Salave S, Gupta R, Patel P, Karunakaran B, Sharma A, Giri J, Benival D, Kommineni N. Chitosan: A potential biopolymer in drug delivery and biomedical applications. Pharm. 2023 Apr 21; 15(4):1313.

Khalaf EM, Abood NA, Atta RZ, Ramírez-Coronel AA, Alazragi R, Parra RM, Abed OH, Abosaooda M, Jalil AT, Mustafa YF, Narmani A. Recent progressions in biomedical and pharmaceutical applications of chitosan nanoparticles: A comprehensive review. Int J Biol Macromol. 2023 Mar 15; 231:123354.

Abd El-Hack ME, El-Saadony MT, Shafi ME, Zabermawi NM, Arif M, Batiha GE, Khafaga AF, Abd El-Hakim YM, Al-Sagheer AA. Antimicrobial and antioxidant properties of chitosan and its derivatives and their applications: A review. Int J Biol Macromol. 2020 Dec 1; 164:2726-44.

Su H, Sun J, Jia Z, Zhao H, Mao X. Insights into promiscuous chitosanases: the known and the unknown. Appl Microbiol Biotechnol. 2022 Nov1; 106(21):6887-6898.

Vanathi P.Production, Optimization and Application of Chitosanase Enzyme From Marine Actinomycetes - Streptomyces Massasporeus. Afr J Bio Sc, 2024 May 6; 6(10).

Vanathi P.Production, Characterization, Application of Chitologiosaccharides Hydrolysate with Partially Purified Chitosanase Enzyme From Marine Isolate Brevidomonas Diminuta.Int J Pharm Res, 2024 Jun 1;15(6): 1835-1844.

Vanathi P, Fernandez S, Shanmugapriya M, et al. Production and characterization of partially purified chitosanase enzyme of Bacillus cereus A4/B4 isolated from biowaste soil samples: bioactive chitooligosaccharide. Food Sci Indian J Res Food Sci Nutr. 2015 Dec 1;2(2):41-48.

Jędrzejczak E, Frąckowiak P, Sibillano T, et al. Isolation and Structure Analysis of Chitin Obtained from Different Developmental Stages of the Mulberry Silkworm (Bombyx mori). Molecules. 2024; Apr 23; 29(9):1914.

Yin N, Du R, Zhao F, Han Y, Zhou Z. Characterization of antibacterial bacterial cellulose composite membranes modified with chitosan or chitooligosaccharide. Carbohydrate Polym. 2020 Feb 1; 229: 115520.

Triunfo M, Tafi E, Guarnieri A, et al. Characterization of chitin and chitosan derived from Hermetia illucens, a further step in a circular economy process. Sci Rep. 2022 Apr 22;12(1):6613.

Boschetti G, Sgarabotto D, Meloni M, et al. Antimicrobial Resistance Patterns in Diabetic Foot Infections, an Epidemiological Study in Northeastern Italy. Antibiotics (Basel). 2021 Qct 13; 10(10):1241.

Silva NS, Araújo NK, Daniele-Silva A, Oliveira JW, Medeiros JM, Araújo RM, Ferreira LD, Rocha HA, Silva-Junior AA, Silva MS, Fernandes-Pedrosa MD. Antimicrobial activity of chitosan oligosaccharides with special attention to antiparasitic potential. Mar drugs. 2021 Feb 12; 19(2):110.

El-Beltagi HS, El-Mahdy OM, Mohamed HI, El-Ansary AE. Antioxidants, Antimicrobial, and Anticancer Activities of Purified Chitinase of Talaromyces funiculosus Strain CBS 129594 Biosynthesized Using Crustacean Bio-Wastes. Agron. 2022 Nov 12; 12(11):2818.

Kowalska-Krochmal B, Dudek-Wicher R. The Minimum Inhibitory Concentration of Antibiotics: Methods, Interpretation, Clinical Relevance. Pathogens. 2021 Feb 4;10(2):165.

Parvekar P, Palaskar J, Metgud S, Maria R, Dutta S. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of silver nanoparticles against Staphylococcus aureus. Biomater Investig Dent. 2020 Jul 23; 7(1):105-109.

Miguez N, Kidibule P, Santos-Moriano P, Ballesteros AO, Fernandez-Lobato M, Plou FJ. Enzymatic synthesis and characterization of different families of chitooligosaccharides and their bioactive properties. Appl Sci. 2021 Apr 3; 11(7):3212.

Wang K, Yu D, Bai Y, Cao H, Guo J, Su Z. Isolation and Purification of Chitosan Oligosaccharides (Mw ≤ 1000) and Their Protective Effect on Acute Liver Injury Caused by CCl4. Mar Drugs. 2024 Mar 8;22(3):128.

Paoletti F, El-Sagheer AH, Allard J, Brown T, Dushek O, Esashi F. Molecular flexibility of DNA as a key determinant of RAD51 recruitment. EMBO J. 2020;39(7):e103002.

de Andrade RC, de Araújo NK, Torres-Rêgo M, Furtado AA, Daniele-Silva A, de Souza Paiva W, de Medeiros Dantas JM, da Silva NS, da Silva-Júnior AA, Ururahy MA, de Assis CF. Production and characterization of chitooligosaccharides: Evaluation of acute toxicity, healing, and anti-inflammatory actions. Inter J Mole Sci. 2021 Sep 30;22(19):10631.

Li J, Tang R, Zhang P, Yuan M, Li H, Yuan M. The Preparation and Characterization of Chitooligosaccharide-Polylactide Polymers, and In Vitro Release of Microspheres Loaded with Vancomycin. J Funct Biomater. 2022 Aug 4;13(3):113.

Mohite P, Shah SR, Singh S, et al. Chitosan and chito-oligosaccharide: a versatile biopolymer with endless grafting possibilities for multifarious applications. Front Bioeng Biotechnol. 2023 May 19; 11:1190879.

Hong T, Yin JY, Nie SP, Xie MY. Applications of infrared spectroscopy in polysaccharide structural analysis: progress, challenge and perspective. Food Chem X. 2021; 12:100168.

Chen X, Wu X, Zhang K, Sun F, Zhou W, Wu Z, Li X. Purification, characterization, and emulsification stability of high-and low-molecular-weight fractions of polysaccharide conjugates extracted from green tea. Food Hydrocoll. 2022 Aug 1; 129:107667.

Campanale C, Savino I, Massarelli C, Uricchio VF. Fourier transform infrared spectroscopy to assess the degree of alteration of artificially aged and environmentally weathered microplastics. Polymers. 2023 Feb 11; 15(4):911.

Fumoto E, Sato S, Takanohashi T. Determination of carbonyl functional groups in heavy oil using infrared spectroscopy. Energy & Fuels. 2020 Jan 24; 34(5):5231-5.

Islam MM, Islam R, Hassan SM, Karim MR, Rahman MM, Rahman S, Hossain MN, Islam D, Shaikh MA, Georghiou PE. Carboxymethyl chitin and chitosan derivatives: synthesis, characterization and antibacterial activity. Carbohydr Polym. 2023 Jun 1;5:100283.

Igcı N, Demiralp FD. A Fourier transform infrared spectroscopic investigation of macrovipera lebetina lebetina and M. l. obtusa Crude Venoms. Eur J Biol. 2020 Jun 1;79(1):14-22.

Giraldo JD, García Y, Vera M, Garrido-Miranda KA, Andrade-Acuña D, Marrugo KP, Rivas BL, Schoebitz M. Alternative processes to produce chitin, chitosan, and their oligomers. Carbohydr Polym. 2024; 332:121924.

Xu T, Sun R, Zhang Y, Zhang C, Wang Y, Wang ZA, Du Y. Recent research and application prospect of functional oligosaccharides on Intestinal disease treatment. Molecules. 2022 Nov 7;27(21):7622.

Xu Y, Wang H, Zhu B, Yao Z. Biochemical characterization and elucidation action mode of a new endolytic chitosanase for efficient preparation of chitosan oligosaccharides. Biomass Convers Bior. 2023 Feb 4:1-9

Hirano S. Applications of chitin and chitosan in the ecological and environmental fields. InApplications of Chitan and Chitosan 2020 Aug 18 (pp. 31-54). CRC Press.

Másson M. Antimicrobial properties of chitosan and its derivatives. In: Chitosan for Biomaterials III: Structure-Property Relationships. Cham: Springer International Publishing; 2021:131-168.

Chandrasekaran M, Kim KD, Chun SC. Antibacterial activity of chitosan nanoparticles: A review. Processes. 2020 Sep 17;8(9):1173.

Li J, Zhuang S. Antibacterial activity of chitosan and its derivatives and their interaction mechanism with bacteria: Current state and perspectives. Eur Polym J. 2020 Sep 5; 138:109984.

Victoria T NS, Kumari T SD, Lazarus B. Antibacterial Activity of Chitosan on Faecal Indicator Bacteria Isolated from Sewage Outfall Nearby Kanyakumari Coast. J Adv Zool. 2024 Apr 1; 45(2).

Sambyal K, Sharma P, Singh RV. Antimicrobial activity of chitooligosaccharides. In: Chitooligosaccharides: Prevention and Control of Diseases. Cham: Springer International Publishing; 2022:301-307

Yan D, Li Y, Liu Y, Li N, Zhang X, Yan C. Antimicrobial properties of chitosan and chitosan derivatives in the treatment of enteric infections. Molecules. 2021 Nov 25; 26(23):7136.

Attjioui M, Gillet D, El Gueddari NE, Moerschbacher BM. Synergistic antimicrobial effect of chitosan polymers and oligomers. Mol Plant Microbe Interact. 2021 Jul 30; 34(7):770-8.

Published

26-06-2024

How to Cite

VANATHI, P. “PRODUCTION AND CHARACTERIZATION OF CHITOOLIGOSACCHARIDE HYDROLYSATE PREPARED FROM CHITOSANASE ENZYME OF MARINE ISOLATES”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 16, no. 08, June 2024, doi:10.22159/ijpps.2024v16i8.51671.

Issue

Section

Original Article(s)