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Abstract
This study aimed to screen Streptomyces strains capable of producing chitinase and to determine the culture conditions affecting the enzyme activity of the most promising strain. Twenty-four Streptomyces isolates were evaluated for chitinase production based on the diameter of the colloidal chitin (CC) hydrolysis zone. The most active strain was identified by 16S rRNA gene sequencing and phylogenetic analysis. Factors influencing chitinase production were investigated, including initial CC concentration (0–3.5%, at 0.5% intervals), pH (4-9), incubation temperature (30, 35, and 40°C), and cultivation time (0–144 h, at 12-h intervals). The crude enzyme was obtained by centrifuging the culture broth (5,000 rpm for 10 minutes at 4 °C) to remove the cells. Chitinase activity in the culture supernatant was determined using a colorimetric assay with 3,5-dinitrosalicylic acid (DNS) reagent at 540 nm. The results indicated that 19 of the 24 tested isolates exhibited chitinase activity, with strain N2R3 producing the largest hydrolysis zone (16.3 mm). Sequence analysis identified this strain as Streptomyces malaysiense. The maximum chitinase activity (3.33 U/mL) was obtained under conditions of 3% CC, pH 8, 30°C, and 84 h of incubation. S. malaysiense N2R3 exhibited strong potential for chitinase applicability in the biotechnological degradation and valorization of chitin-containing by-products.
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