A new alkaline keratinase, which could kill Meloidogyne incognita (a root-knot nematode) was separated and purified from Bacillus sp. 50-3 in this study. The solid ammonium sulfate was selected to precipitate the enzyme and its proper adding mass was also determined. After solid ammonium sulfate precipitation and liquid chromatography on DEAE-Sephadex-A50 column, there was 17.7-fold purification with a yield of 46.5%, as determined by azokeratin as substrate. The purification effect was determined through SDS-PAGE and the molecular weight of the enzyme was found to be 27,423 Da by the MALDI-TOF-MS. When the secondstage juveniles of Meloidogyne incognita were exposed to 50 lg/ml of keratinase solution, 98.5% of Meloidogyne incognita mortality rates were obtained compared to control after 24 h. Its simple purification step and high yield from the cheap medium affords this keratinase great biotechnological potential, especially in controlling rootknot nematodes such as Meloidogyne incognita. To the best of our knowledge, this study is the first report that uses keratinase as a pesticide.

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An amperometric horseradish peroxidase (HRP) biosensor based on multilayer films containing carbon nanotubes (CNTs) and HRP was developed. With the pH regulation of the dispersion solution of CNTs, the sensitivity of the HRP multilayer film biosensor is tunable by the control of the dissociation of CNTs. The successful formation of multilayers was confirmed by UV-visible spectroscopy. The features of multilayers were characterized by SEM and electrochemical impedance spectrum (EIS). The performance of the HRP biosensor is reported for the amperometric detection of phenols. The biosensor presented a linear response for catechol from 9.1 × 10 -8 -6.45 × 10 -5 mol/L, with a sensitivity of 0.00554 A·L/mol and a detection limit of 8.5 × 10 -8 mol/L. The study can provide a feasible simple approach for developing a new sensitivity tunable method for CNTs-based biosensors.

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