Chitinase production by an Arthrobacter sp. lysing cells of Fusarium roseum

An Arthrobacter sp. which actively lysed Fusarium roseum was found to liberate chitinase (E.C. 3.2-1.14, chitin glycanohydrolase), an enzyme essential for the hydrolysis of chitin, a major component of fusarial hyphal walls. Factors involved in the production of chitinase were investigated by modifying culture conditions and assaying for enzyme activity. Production occurred on colloidal chitin as well as on native chitin supplemented with yeast extract or peptone. Enzyme production paralleled growth; liberation of enzyme took place during the log phase with the maximum yield being obtained at the stationary phase. Addition of the non-ionic surfactant, polyoxyethylene sorbitan monooleate (Tween 80) increased enzyme yield. An inverse relationship was found between the amount of enzyme produced and the quantity of n-acetyl-glucosamine liberated. The enzyme was generally not produced when grown on various other carbohydrates. These findings suggest that chitinase is inducible and that chitin breakdown is regulated by a repressor-inducer mechanism.Initial hydrolysis rates of colloidal chitin were proportional to the concentration of chitinase used. Optimal pH and temperature for enzyme activity were 4.9 and 50°C, respectively. Purification of the chitinase was obtained by (NH₄)₂SO₄ precipitation followed by DEAE-cellulose and Sephadex chromatography, achieving a 12-fold increase in specific activity.