重组里氏木霉产漆酶及其对染料金橙Ⅱ的脱色

doi:10.3785/j.issn.1008-973X.2014.10.024

浙江大学学报(工学版)

生物医学工程、化学工程

重组里氏木霉产漆酶及其对染料金橙Ⅱ的脱色

陶亮亮, 赵杰, 夏黎明

浙江大学生物质化工教育部重点实验室,浙江杭州 310027

Laccase production by recombinant Trichoderma reesei and its application for decolorization of gold orangeⅡ

TAO Liang-liang, ZHAO Jie, XIA Li-ming

Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang University,

Hangzhou 310027, China

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摘要：

对已有的10个重组里氏木霉转化子进行产酶试验,从中筛选到一个高产转化子,摇瓶发酵96 h,漆酶活力可以达到8.85 IU/mL.SDS-PAGE结果表明：该重组漆酶的相对分子质量约为68 000.重组漆酶的最适温度为65 ℃,该酶具有较好的耐热性能,经70 ℃处理60 min后残余相对酶活维持在70%以上；最适pH为2.2,在pH为2.6～4.2的条件下具有良好的稳定性.采用发酵粗酶液对100 mg/L的偶氮染料金橙Ⅱ进行脱色试验,对影响脱色效果的主要因子进行研究.结果表明,在温度为50 ℃、pH为3.5、介体HOBT用量为2 mmol/L、漆酶用量为0.2 IU/mL的条件下作用5 h,金橙Ⅱ的脱色率可以达到90.6%.

Abstract:

A transformant of Trichoderma reesei with high production of laccase was screened from 10 recombinant transformants by fermentation experiment, and the maximum laccase activity was achieved at 96 h (8.85 IU·mL-1) in shaking flask. SDS-PAGE analysis showed that the recombinant laccase had a relative molecular mass of around 68 000. The optimum temperature and pH value for the recombinant laccase reaction were 65 ℃ and 2.2, respectively. The laccase was stable at pH between 2.6 and 4.2. Approximately 70% of the initial enzyme activity was retained after incubation of the laccase at 70 ℃ for 60 min, indicating that the laccase was highly thermostable. The effects of primary factors on the decolorization of gold orangeⅡ（100 mg.L-1）were analyzed and the decolorization rate reached 90.6% in 5 h at 50 ℃, pH 3.5, HOBT concentration 2 mmol·L-1 and laccase dosage 0.2 IU·mL-1. Excellent thermostability and efficient decolorization suggest that this crude recombinant laccase can be effectively applied to decolorize the recalcitrant azo dyes from effluents.

出版日期: 2014-10-01

Q 814

基金资助:

浙江省重点科技创新团队计划资助项目（2011R50002）.

通讯作者: 夏黎明,男,教授,博导 E-mail: xialm@zju.edu.cn

作者简介: 陶亮亮（1989—），男，硕士生，从事生物化工的研究. E-mail: leonfang1314@163.com

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引用本文:

陶亮亮, 赵杰, 夏黎明. 重组里氏木霉产漆酶及其对染料金橙Ⅱ的脱色[J]. 浙江大学学报(工学版), 10.3785/j.issn.1008-973X.2014.10.024.

TAO Liang-liang, ZHAO Jie, XIA Li-ming.

Laccase production by recombinant Trichoderma reesei and its application for decolorization of gold orangeⅡ

. JOURNAL OF ZHEJIANG UNIVERSITY (ENGINEERING SCIENCE), 10.3785/j.issn.1008-973X.2014.10.024.

链接本文:

http://www.zjujournals.com/eng/CN/10.3785/j.issn.1008-973X.2014.10.024 或 http://www.zjujournals.com/eng/CN/Y2014/V48/I10/1879

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