Abstract
The production of inexpensive chitinolytic enzymes is an element in the utilization of shellfish-processing waste. In this study, shrimp and crab shell powder, prepared by treating shrimp- and crab-processing waste by boiling and crushing, was used as a substrate for the isolation of an antifungal chitinase-producing microorganism. Bacillus subtilis NPU 001, a strain isolated from soil samples, excreted a chitinase when cultured in a medium containing 2% (w/v) shrimp and crab shell powder as the major carbon source. The chitinase, which was purified by sequential chromatography, had a Mw of 31 kDa and a pI of 5.4. The purified chitinase (2 mg ml−1) inhibited hyphal extension of the fungus Fusarium oxysporum. Compared with other known bacterial chitinases, the unique characteristics of NPU 001 chitinase include antifungal activity against plant-pathogenic fungi and the production of chitotriose as the major enzymatic hydrolysate from colloidal chitin.
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References
Chang WT, Chen C, Wang SL (2003) An antifungal chitinase produced by Bacillus cereus using shrimp and crab shell powder as a carbon source. Curr Microbiol 47:102–108
Chang WT, Chen YC, Jao CL (2007) Antifungal activity and enhancement of plant growth Bacillus cereus grown on chitin wastes. Biores Technol 98:1226–1230
Chang WT, Hsieh CH, Hsieh HS, Chen C (2009) Conversion of crude chitosan to an anti-fungal protease by Bacillus cereus. World J Microbiol Biotechnol 25:375–382
Deane EE, Whipps JM, Lynch JM, Peberdy JF (1998) The purification and characterization of a Trichoderma harzianum exochitinase. Biochem Biophys Acta 1383:101–110
Di Pietro A, Lorito M, Hayes CK, Broadway RM, Harman GE (1993) Endochitinase from Gliocladium virens: isolation, characterization, and synergistic antifungal activity in combination with gliotoxin. Phytopathology 83:308–313
Imoto T, Yagishita K (1971) A simple activity measurement by lysozyme. Agric Biol Chem 35:1154–1156
Kishore GK, Pande S (2007) Chitin-supplemented foliar application of chitinolytic Bacillus reduces severity of Botrytis gray mold disease in chickpea under controlled conditions. Lett Appl Microbiol 44:98–105
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–688
Lorito M, Harman GE, Hayes CK, Broadway RM, Tronsmo A, Woo SL, Di Pietro A (1993) Chitinolytic enzyme produced by Trichoderma harzianum: Antifungal activity of purified endochitinase and chitobiosidase. Mol Plant Pathol 83:302–307
Patil RS, Ghormade V, Deshpande MV (2000) Chitinolytic enzymes: an exploration. Enzyme Microb Technol 26:473–483
Reyes-Ramirez A, Escudero-Abarca BI, Aguilar-Uscanga G, Hayward-Janes PM, Barboza-Corona JE (2004) Antifungal activity of Bacillus thuringiensis chitinase and its potential for the biocontrol of phytopathogenic fungi in soybean seeds. J Food Sci 69:M131–M134
Sakuda S (1996) Studies on the chitinase inhibitors. Allosamidins. In: Muzzarelli RAA (ed) Chitin enzymology, vol 2. Eur Chitin Soc, Grottammare, Italy, pp 203–212
Shaikh SA, Deshpande MV (1993) Chitinolytic enzymes: their contribution to basic and applied research. World J Microbiol Biotechnol 9:468–475
Wang SL, Chang WT (1997) Purification and characterization of two bifunctional chitinase/lysozymes extracellularly produced by Pseudomonas aeruginosa K-187 in shrimp and crab shell powder medium. Appl Environ Microbiol 63:380–386
Acknowledgments
This work was supported in part by a grant from the National Science Council, Republic of China (NSC97-2221-E-346-005).
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Chang, WT., Chen, ML. & Wang, SL. An antifungal chitinase produced by Bacillus subtilis using chitin waste as a carbon source. World J Microbiol Biotechnol 26, 945–950 (2010). https://doi.org/10.1007/s11274-009-0244-7
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DOI: https://doi.org/10.1007/s11274-009-0244-7