端梗霉Z45产漆酶培养基的优化及其对染料的脱色

Medium Optimization for Laccase Production by Acrophialophora sp.Z45 and Its Decolorization of Dyes

菌株Acrophialophora sp.Z45是一株产漆酶的端梗霉属真菌,本文依据不同培养基中菌株Z45对愈创木酚的氧化圈大小探究了影响该菌株产漆酶的因素并设计正交试验优化其产漆酶培养基,进而比较了菌株Z45在土豆葡萄糖培养基和优化后的产漆酶培养基中的漆酶活力差异,最后基于漆酶与染料脱色的相关性评价了菌株Z45对8种合成染料的脱色能力。结果表明,单因素及正交试验确定了菌株Z45优化后的产漆酶培养基为:基础产酶培养基中以蔗糖为碳源、硝酸钠为氮源、C/N比为45∶1、pH为5.0;在优化后的产漆酶培养基中菌株Z45的漆酶活性显著提高;菌株Z45对溴酚蓝、中性红、亚甲基蓝、甲基蓝和结晶紫等5种染料均有一定的脱色能力,其中对三苯甲烷染料甲基蓝的脱色能力最强,菌株Z45的粗酶液能使甲基蓝快速脱色。在较低甲基蓝浓度的固体培养基上,甲基蓝完全脱色的时间不受染料浓度的影响;而较高甲基蓝浓度下,甲基蓝完全脱色的时间随染料浓度升高逐渐延长。

The strain Acrophialophora sp.Z45 is a laccase-producing fungus,which was discovered in the previous study of our laboratory.In this study,based on the size of guaiacol-oxidation zone on different medium,the factors influencing the laccase production by strain Z45 were studied.The orthogonal experiments were designed to optimize the basic medium for laccase production and the difference in laccase activity before and after medium optimization was then compared.Finally,the ability of strain Z45 to decolorize eight synthetic dyes was evaluated based on the correlation between laccase and dye decolorization.The results based on the single factor experiment and orthogonal test showed that the optimized medium for laccase production was as follows:sucrose as carbon source,sodium nitrate as nitrogen source,the carbon to nitrogen ratio was 45∶1,with a pH value of 5.0.The laccase activity of strain Z45 was significantly improved after medium optimization.Strain Z45 had certain decolorization ability to bromophenol blue,neutral red,methylene blue,methyl blue and crystalline violet,among which the decolorization ability to triphenylmethane dye methyl blue was the strongest.The crude enzyme solution of strain Z45 decolorized methyl blue rapidly.On the solid medium with lower concentrations of methyl blue,the time for complete decolorization of methyl blue was not affected by the dye concentration,but at higher concentrations of methyl blue,the time for complete decolorization was gradually extended with the increase of dye concentration.