球形芽孢杆菌C3-41菌株芽孢漆酶的提
发布时间：2019-05-29 05:55
【摘要】：目前对漆酶的研究主要集中在白腐真菌漆酶上,但随着对细菌漆酶研究的增多,结果发现,细菌漆酶与白腐真菌漆酶相比而言,细菌漆酶具有耐高温、pH耐受度高、来源丰富、容易在工业宿主生物中表达便于实现大规模工业化生产等优点,由芽孢杆菌产生的芽孢漆酶更具有最适温度高、热稳定性强、pH适用范围宽,碱性耐受度高等特点,使其具有广泛的应用前景。(1)芽孢杆菌C3-41产漆酶性质的初步探究结果显示,常用漆酶检测体系中,pH4时0.1mol/L磷酸氢二钠-柠檬酸缓冲溶液1.8mL、0.6mM/L ABTS 0.2mL以及1mL漆酶样品所组成的3mL缓冲体系为该芽孢漆酶的最佳检测体系。三种处理方式不同的样品的漆酶活性和蛋白含量的比较即可得知球形芽孢杆菌C3-41产生漆酶的主要部位是芽孢,且该芽孢漆酶为胞内酶。(2)芽孢杆菌C3-41芽孢漆酶的酶学性质研究显示,pH对芽孢漆酶活性影响比较明显,当pH为4时芽孢漆酶活性最大;该芽孢漆酶的温度稳定性较好,在较高的温度的下酶活仍然维持在较高的水平上,其最佳温度高达50℃;在体系中金属离子终浓度为0.2mmol/L时,Fe2+、Fe3+均对漆酶活性具有抑制,Cu2+、Zn2+、Ba2+、Ca2+对漆酶活性均具有促进作用,其中Ca2+的促进作用最明显;添加工作浓度的DTT、β-巯基乙醇、EDTA、PMSF四种常用蛋白酶抑制剂发现,DTT和β-巯基乙醇对其酶活的抑制作用较明显,PMSF的抑制作用最微弱。(3)芽孢杆菌C3-41芽孢漆酶对活性艳蓝KN-R、活性艳红X-3B、孔雀绿三种染料的脱色特性研究显示,在染料浓度不超过2g/L情况下,该芽孢漆酶对活性艳蓝的脱色率随染料浓度增大而增大,最大浓度时脱色率为87.8%;对活性艳红的脱色率随染料浓度增大而减小,最大浓度时脱色率为72.8%;对孔雀绿的脱色率随染料浓度变化不大,除浓度达到2g/L时脱色率为82.2%外,其他浓度脱色率均大于86%。(4)芽孢漆酶与纯化漆酶相比,芽孢漆酶的温度耐受度和pH耐受度更高本论文研究结果表明,球形芽孢杆菌C3-41芽孢漆酶对染料脱色、漆酶酶学特性以及以及工程应用中降低成本方面均有较好的利用价值。
[Abstract]:At present, the research on laccase is mainly focused on white rot fungus laccase, but with the increase of bacterial laccase, the results show that bacterial laccase has high temperature resistance, pH tolerance and abundant sources compared with white rot fungus laccase. It is easy to express in industrial host organisms to realize large-scale industrial production. The spore laccase produced by Bacillus has the characteristics of high optimum temperature, strong thermal stability, wide application range of pH and high alkaline tolerance. It has a wide range of application prospects. (1) the preliminary study on the properties of laccase produced by Bacillus C3341 shows that 0.1mol/L disodium hydrogen phosphate citric acid buffer solution 1.8 mL in common laccase detection system, The 3mL buffer system composed of 0.6mM/L ABTS 0.2mL and 1mL laccase samples is the best detection system for the spore laccase. By comparing the laccase activity and protein content of the samples with different treatment methods, it can be found that the main part of laccase produced by Bacillus globularis C _ 3 / 41 is spores. The spore laccase was intracellular enzyme. (2) the enzyme properties of Bacillus C3o41 spore laccase showed that pH had obvious effect on spore laccase activity, and when pH was 4, spore laccase activity was the highest. The temperature stability of the spore laccase is good, and the enzyme activity is still maintained at a high level at higher temperature, and the optimum temperature is as high as 50 鈩，
本文编号：2487706
[Abstract]:At present, the research on laccase is mainly focused on white rot fungus laccase, but with the increase of bacterial laccase, the results show that bacterial laccase has high temperature resistance, pH tolerance and abundant sources compared with white rot fungus laccase. It is easy to express in industrial host organisms to realize large-scale industrial production. The spore laccase produced by Bacillus has the characteristics of high optimum temperature, strong thermal stability, wide application range of pH and high alkaline tolerance. It has a wide range of application prospects. (1) the preliminary study on the properties of laccase produced by Bacillus C3341 shows that 0.1mol/L disodium hydrogen phosphate citric acid buffer solution 1.8 mL in common laccase detection system, The 3mL buffer system composed of 0.6mM/L ABTS 0.2mL and 1mL laccase samples is the best detection system for the spore laccase. By comparing the laccase activity and protein content of the samples with different treatment methods, it can be found that the main part of laccase produced by Bacillus globularis C _ 3 / 41 is spores. The spore laccase was intracellular enzyme. (2) the enzyme properties of Bacillus C3o41 spore laccase showed that pH had obvious effect on spore laccase activity, and when pH was 4, spore laccase activity was the highest. The temperature stability of the spore laccase is good, and the enzyme activity is still maintained at a high level at higher temperature, and the optimum temperature is as high as 50 鈩，
本文编号：2487706
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