Production and Screening of Streptomyces-Extracellular Chitinase

  • Ibrahim Sani Department of Biochemistry, Faculty of Life Sciences, Kebbi State University of Science and Technology, Aliero, Kebbi State, Nigeria
  • Aminu Argungu Umar Department of Biochemistry, Faculty of Life Sciences, Kebbi State University of Science and Technology, Aliero, Kebbi State, Nigeria
  • Evelyn Uzoamaka Udeze Department of Biochemistry, Faculty of Life Sciences, Kebbi State University of Science and Technology, Aliero, Kebbi State, Nigeria
Keywords: Streptomyces, Soil bacteria, Chitinase, Antifungal, Candida albicans


The aim of this research was to produce Streptomyces-extracellular chitinase and screen its antifungal activity on a clinically isolated Candida albicans. The Streptomyces were isolated from an agricultural farmland; they were identified and screened for the chitinase production. Effects of time, temperature, pH and nitrogen sources on the chitinase production were determined using standard methods. Ammonium sulphate precipitation was used to partially purify the chitinase. Protein concentrations were determined spectrophotometrically using bovine serum albumin as standard. Agar-well diffusion method was used to evaluate the antifungal activity of the chitinase on C. albicans. The isolated Streptomyces were of three (3) strains, and all the strains are Gram positive, catalase positive, oxidase positive while, Strain A and C are indole positive and only Strain B is citrate positive. The maximum chitinase production was at 72 h, 40°C and when yeast extract was used as the nitrogen source. Ammonium sulphate (80%) precipitation yielded the highest enzyme activity of 39.0U/ml. The maximum enzyme activity was observed at temperature of 40oC, pH 5.5 and 1.0% colloidal chitin (substrate). The partially purified chitinase showed a zone of inhibition of 20.11 ± 1.26 mm against the Candida albicans. This result has no significant difference (P>0.05) when compared with that of the standard drug (Fluconazole) with 21.42 ± 0.08 mm zone of inhibition. These findings suggest that Streptomyces at favourable conditions produce chitinase, and this enzyme can be used as an antifungal agent on Candida albicans and other chitin containing fungi.


Al-Ahmadi, K. J., Yazdi, M. T., Najafi, M. F., Shahverdi, A. R., Faramarzi, M. A., Zarrini, G., and Behravan, J. (2008). Optimization of medium and cultivation conditions for chitinase production by the newly isolated: Aeromonas sp. Biotechnology, 7(2), 266-272.

Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 72(1-2), 248-254.

Brzezinska, M. S., Jankiewicz, U., Burkowska, A., and Walczak, M. (2014).Chitinolytic microorganisms and their possible application in environmental protection. Current microbiology, 68(1), 71-81.

Choi, I., Troyer, D. L., Cornwell, D. L., Kirby-Dobbels, K. R., Collante, W. R., and Simmen, F. A. (1997). Closely related genes encode developmental and tissue isoforms of porcine cytochrome P450 aromatase. DNA and cell biology, 16(6), 769-777.

El‐Tarabily, K. A., Soliman, M. H., Nassar, A. H., Al‐Hassani, H. A., Sivasithamparam, K., McKenna, F., and Hardy, G. S. J. (2000). Biological control of Sclerotinia minor using a chitinolytic bacterium and actinomycetes. Plant Pathology, 49(5), 573-583.

Gomaa, E. Z. (2012). Chitinase production by Bacillus thuringiensis and Bacillus licheniformis: their potential in antifungal biocontrol. The Journal of Microbiology, 50(1), 103-111.

Hiroshi, T., Kazuhiro, F., Hiromi, T., Katsushiro, M., Chiaki, I., Yoshiro, O., and Yoshihiko, I. (1994). Gene sequence, purification and characterization of N-acetyl-β-glucosaminidase from a marine bacterium, Alteromonas spp. strain O-7. Gene, 146(1), 111-115.

Hoster, F., Schmitz, J. E., and Daniel, R. (2005). Enrichment of chitinolytic microorganisms: isolation and characterization of a chitinase exhibiting antifungal activity against phytopathogenic fungi from a novel Streptomyces strain. Applied Microbiology and Biotechnology, 66(4), 434-442.

Hussein, R. H. M. (2013). The Influence of Camel and Cattle Meat Quality and Chemical Composition on the Total Microbial Load and Antimicrobial Activities (Doctoral dissertation, University of Gezira).

in vitro. Proceedings of the National Academy of Sciences, 73(12), 4570-4574.

Joo, G. J. (2005). Purification and characterization of an extracellular chitinase from the antifungal biocontrol agent Streptomyces halstedii. Biotechnology letters, 27(19), 1483-1486.

Khan, S. T., Komaki, H., Motohashi, K., Kozone, I., Mukai, A., Takagi, M., and Shin‐ya, K. (2011). Streptomyces associated with a marine sponge Haliclona spp.; biosynthetic genes for secondary metabolites and products. Environmental microbiology, 13(2), 391-403.

Korn-Wendisch, F., and Kutzner, H. J. (1991).The family Streptomycetaceae. The Prokaryotes. A handbook on the biology of bacteria: ecophysiology, isolation, identification, applications, Chap. 41.

Kumar, R. R., and Jadeja, V. J. (2016). Isolation of actinomycetes: A complete approach. International Journal of Current Microbiology Applied Science, 5, 606-18.

Limón, M. C., Pintor-Toro, J. A., and Benítez, T. (1999).Increased antifungal activity of Trichoderma harzianum transformants that overexpress a 33-kDa chitinase. Phytopathology, 89(3), 254-261.

Lu, Y., Zen, K. C., Muthukrishnan, S., and Kramer, K. J. (2002).Site-directed mutagenesis and functional analysis of active site acidic amino acid residues D142, D144 and E146 in Manduca sexta (tobacco hornworm) chitinase. Insect biochemistry and molecular biology, 32(11), 1369-1382.

Mitchell, S. (2019). An integrative approach to understanding antimicrobial resistance in New Zealand (Doctoral dissertation, University of Otago).

Narayana, K. J., and Vijayalakshmi, M. (2009).Chitinase production by Streptomyces sp. ANU 6277. Brazilian Journal of Microbiology, 40(4), 725-733.

Nawani, N. N., and Kapadnis, B. P. (2004). Production dynamics and characterization of chitinolytic system of Streptomyces sp. NK1057, a well-equipped chitin degrader. World Journal of Microbiology and Biotechnology, 20(5), 487-494.

Nawani, N. N., B. P. Kapadnis, A. D. Das, A. S. Rao, and S. K. Mahajan.(2002): "Purification and characterization of a thermophilic and acidophilic chitinase from Microbispora sp. V2." Journal of Applied Microbiology 93, no. 6(2): 965-975.

Oberlander, H., and Silhacek, D. L. (1998).New perspectives on the mode of action of benzoylphenyl urea insecticides. In Insecticides with Novel Modes of Action (pp. 92-105).Springer, Berlin, Heidelberg.

Oyeleke, S. B., and Jibrin, N. M. (2009).Production of bioethanol from guinea cornhusk and millet husk. African Journal of Microbiology Research, 3(4), 147-152.

Palmer, E., and Freeman, T. (2004). Investigation into the use of C-and N-terminal GFP fusion proteins for subcellular localization studies using reverse transfection microarrays. International Journal of Genomics, 5(4), 342-353.

Rattanakit, N., Plikomol, A., Yano, S., Wakayama, M., and Tachiki, T. (2002). Utilization of Shrim Shellfish Waste as a Substrate for Solid-State Cultivation of a Culture Based on Chitinase Formation Which Is Necessary for Chitin Assimilation. Journal of Bioscience and Bioengineering, 93(6), 550-556.

Shanmugaiah, V., Mathivanan, N., Balasubramanian, N., and Manoharan, P. T. (2008). Optimization of cultural conditions for production of chitinase by Bacillus laterosporous MML2270 isolated from rice rhizosphere soil. African Journal of Biotechnology, 7(15).

Sowmya, B., Gomathi, D., Kalaiselvi, M., Ravikumar, G., Arulraj, C., and Uma, C. (2012).Production and Purification of Chitinase by Streptomyces sp. from Soil.Journal of Advanced Scientific Research, 3(3).

Suresh, G., Raghu, P., Prakash, U., Suresh, T. C., Kumaran, S., Bharathi, S., ...& Poonguzhali, T. V. (2018). Screening of the Antibiotic Resistant Environmental Bacteria Isolated from Selected Niches in and Around Kanchipuram Town, India. International Journal of Current Microbiology and Applied Sciences, 7(6), 3665-3674.

Taechowisan, T., Peberdy, J. F., and Lumyong, S. (2003). Chitinase production by endophytic Streptomyces aureofaciens CMUAc130 and its antagonism against phytopathogenic fungi. Annals of microbiology, 53(4), 447-462.

Wang, X., Ding, X., Gopalakrishnan, B., Morgan, T. D., Johnson, L., White, F. F., and Kramer, K. J. (1996). Characterization of a 46 kDa insect chitinase from transgenic tobacco. Insect Biochemistry and Molecular Biology, 26(10), 1055-1064.

Zhang, J., Kopparapu, N. K., Yan, Q., Yang, S., and Jiang, Z. (2012). Purification and characterisation of a novel chitinase from persimmon (Diospyros kaki) with antifungal activity. Food chemistry, 138(2-3), 1225-1232.

How to Cite
Sani, I., Umar, A. A., & Udeze, E. U. (2021). Production and Screening of Streptomyces-Extracellular Chitinase. Journal La Lifesci, 2(4), 1-16.