Abstract

ObjectivesThermal and pH stabilities are extremely important for the application of pectate lyase. In this study, we aimed to obtain a strain that is able to produce pectate lyase with good pH stability. MethodsIn this study, screening for pectate lyase was performed using plate assays. Fermentation parameters for pectate lyase production were optimized utilizing a single variable optimization. To get insight into the pectate lyase, its enzyme property, purification, identification, and application were performed. ResultsBacillus altitudinis CAS-WZS-08, producing pectate lyase with good pH stability, was isolated. The optimal fermentation conditions of CAS-WZS-08 are 4 g/L pectin, 20 g/L yeast extract, 2% inoculum size, pH 7.0, and 33 °C, which the production of pectate lyase can reach up to 0.71 ± 0.001 U/mL. The optimal pH and temperature of the pectate lyase were 10.0 and 60 °C, respectively. Stored at 4 °C, the pectate lyase was able to keep its full enzyme activity for 24 h under a wide range of pH (4.0–10.0) condition. With pH 10.0, this pectate lyase was stable under 30-45 °C. In addition, it can be activated by Mn2+, Cu2+, Co2+, and Ca2+, while inhibited by Fe3+, Ba2+, and Mg2+. Later, the electrophoretic pure protein was acquired through ammonium sulfate precipitation, cation exchange column, and Sephadex G-75. Liquid chromatography tandem-mass spectrometry (LC/MS-MS) further confirmed that the purified protein was pectate lyase with a molecular weight of ∼ 40 kDa. At last, the result of pectate lyase in extracting apple juice demonstrated that it has an excellent juice extraction ability. ConclusionThis study provides an excellent pH-stable pectate lyase with good thermal stability that is a potential candidate for industrial applications.

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