The temperature stability of this enzyme was assayed from the residual exercise after the incubation at various temperatures (20~70 °C) at pH 7.zero for 60 min. The enzyme was purified from tradition supernatant of strain CJ-5. Cell free tradition broth was saturated with ammonium sulfate between 30% and sixty five% concentrations. The solution was left overnight at four °C, centrifuged, and the precipitates had been dissolved in McIlvaine buffer (pH 7.0).
Moreover, the power of Sphingomonas sp. CJ-5 to provide most chitinase activity under delicate circumstances of agitation and aeration could be an necessary benefit in view of additional course of improvement.
The chitinase and chitosanase from Sphingomonas sp. CJ-5 could also be probably useful for industrial application and recycling of chitin wastes significantly. The thermal stability of the chitosanase was larger than that of the chitinase. The pH stability of chitinase or chitosanase was assayed from the residual activity after the incubation at 36 °C or 56 °C within the buffer of varied pH (3~9) for 60 min.
The chitinase and chitosanase of Sphingomonas sp. CJ-5 are fairly steady and active, and degrade chitin effectively, although the enzyme yield isn't high.
The flow fee was maintained at zero.25 ml/min. All purification steps have been done at four °C. The lively fractions comparable to chitinase and chitosanase have been collected and used as supply enzymes for enzyme evaluation.
https://enzymes.bio/ was dialyzed against the same buffer for 24 h, and then subjected to Sephadex G-200 gel filtration column (1.6 cm×60 cm) equilibrated with the identical buffer. The protein was eluted with the same buffer at a circulate fee of 24 ml/h. Each fraction was collected, and the chitinase and chitosanase actions had been measured. The fractions with chitinase or chitosanase activity had been individually pooled, concentrated and additional applied on DEAE-Sepharose Fast Flow column (1.6 cm×20 cm) pre-equilibrated with 10 mmol/L Tris-HCl (pH 7.6). The absorbed chitinase or chitosanase was eluted by a linear gradient of NaCl from zero to zero.5 mol/L in the identical buffer.