Isolation, identification of a laccase-producing fungal strain and enzymatic properties of the laccase
A new type of thermostable laccase was isolated from Paraphoma sp. GZS18, and its partial enzymatic properties were determined. A strain GZS18 of laccase with high yield was screened from forest soil and identified as Paraphoma sp. GZS18 through morphological characteristics and ITS sequence analysis. The laccase of Paraphoma sp. GZS18 (Lac-P) was obtained through cation–anion exchange chromatography, gel filtration chromatography, and other purification processes. The testing result shows that Lac-P is a single protein of 75 kDa, and the 11 amino acid sequences in the N-terminal are AXaVSVASREMT (Xa was the non-standard protein). The optimum temperature and optimum pH of lac-P activity are substrate-independent. The temperature is in the range of 50–70 °C, and pH has high catalytic efficiency in the acidic range. Lac-P has good stability in the temperature and pH. The half time at 70–60 °C is 1.5 and 4 h, respectively. At pH 6–9 and room temperature, there is more than 80% activity 24 h later. Lac-P is tolerant of most metal ions and low concentrations of inhibitors but is inhibited by Hg2+, Fe2+ and NaN3. The laccase from Paraphoma sp. GZS18 at high temperature and pH 6–9, with strong stability, has better industrial application characteristics.
KeywordsCharacterization Laccase Paraphoma sp. Purification
This work was financially supported by the Science technology Program of traditional Chinese medicine and minority medicine in Guizhou Province of traditional Chinese medicine Administration (No. QZYY-2014-028); the Doctor Foundation of Binzhou University (Nos. 2016Y02 and 2016Y17); and the Natural Science Foundation of Shandong Province (Nos. ZR2016CL01 and ZR2016BL16).
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Conflict of interest
The author(s) declare(s) that there is no conflict of interest regarding the publication of this article.
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