漆酶/HOBT体系对苯胺蓝的脱色条件及机理研究

研究了Pleurotus sp.N3固态发酵所得漆酶对三芳基甲烷类染料苯胺蓝的脱色条件和机理.通过可见光光谱扫描和非变性凝胶电泳的分析发现漆酶能够对苯胺蓝进行脱色,介体可以显著增强脱色效果,提高脱色率.分别考察了介体、酶活和温度对苯胺蓝脱色的影响,结果表明,在酶活为200 U/L、1-羟基苯并三唑(HOBT)浓度为4mmol/L、温度为40℃的条件下,反应6h后苯胺蓝(24 mg/L)的脱色率达到90.42％.对脱色0、4h、11h和23 h的产物进行了超高效液相色谱-质谱(UPLC-MS)分析,推断出漆酶/HOBT体系对苯胺蓝的脱色过程中有显色基团碳氮双键(C=N)的断裂.     

镰刀菌(Fusarium sp.)HJ01对中性艳蓝GL的脱色降解特性研究

利用镰刀菌(Fusarium sp.)HJ01降解中性艳蓝GL,经扩增翻译延伸因子TEF1对HJ01菌株做了种的鉴定,并考察了温度、pH值对该菌株降解酶漆酶的活性,以及对中性艳蓝GL脱色率及COD去除率的影响.结果表明,该菌株属层出镰刀菌(Fusarmm proliferatum).HJ01菌株所产降解酶漆酶在pH为2.0～8.0内相对活性在12%～96%之间,在pH=4.0时,相对活性最大;在25～65 ℃内,随着温度的升高,漆酶相对活性增大;在65～85 ℃内,随着温度增大,漆酶相对活性降低.HJ01菌株在25℃、50 r·min-1条件下,于液体培养基中培养96h后加入到25mL含100mg·L-1中性艳蓝GL的溶液中,在35℃、50 r·min-1、pH=4.0条件下继续培养100h后,中性艳蓝GL的脱色率达100%,培养至120h后,COD去除率达87.8%.紫外光谱分析表明,中性艳蓝GL的发色基团蒽醌环被漆酶打破.     

一株白腐菌产生的漆酶对RB亮蓝的脱色作用

W 1是一株能在液体条件下产漆酶的白腐菌 ,纯化的漆酶对RB亮蓝有很好的脱色作用 .漆酶的最适脱色温度为 4 5℃ ,最适脱色pH值为 6 .0 ,脱色pH范围在 4～ 7之间 .当溶液中漆酶活力为 2 .0× 10 3 U/L时 ,在最适脱色条件下、16h内 ,RB亮蓝 (30 0mg/L)的脱色率可以达到 90 % .经酶作用后 ,RB亮蓝在 4 30～ 70 0nm范围内的特征颜色吸收峰基本消失 .实验证明 ,在相同的条件下 ,漆酶粗酶对RB亮蓝有更好的脱色效果 .图 7表 1参 6     

处理秸秆纤维素乙醇废水的东北土著白腐真菌筛选及特性研究

为了提高秸秆纤维素乙醇废水的处理效果,选择6种东北土著白腐真菌,对2%的秸秆纤维素乙醇废水中木质素进行降解处理.采用正交试验法对筛选出的高效降解菌进行产漆酶培养基的优化.结果表明:6种白腐真菌最高酶活大小顺序为青顶拟多孔菌>血红密孔菌>糙皮侧耳菌>彩绒革盖菌>烟色烟管菌>灵芝;血红密孔菌、糙皮侧耳菌、彩绒革盖菌、青顶拟多孔菌、灵芝、烟色烟管菌在第0天起始质量浓度为640.9~716.6 mg/L,在第14天木质素的质量浓度分别为434.0、411.2、441.8、441.7、533.3、503.5 mg/L,对木质素的去除率分别为37.1%、37.0%、31.8%、31.7%、25.6%、21.4%,并分别在第12、12、4、4、2、6天木质素降解趋于平稳,表明降解效果最好的菌种为血红密孔菌.血红密孔菌产漆酶培养基最优组合方案:最佳碳源为锯末,质量浓度为35 g/L;最佳氮源为蛋白胨,质量浓度为4 g/L;最佳pH为5.极差分析表明,各因素对血红密孔菌产漆酶的影响顺序为碳源>氮源> pH >氮源质量浓度>碳源质量浓度.在最佳培养基条件下,废水中木质素降解率达41.1%.研究显示,血红密孔菌可以作为生物法处理秸秆纤维素乙醇废水的菌种资源,也可为今后的进一步应用研究提供科学依据.      

木质素降解酶研究进展与外生菌根真菌

综述了木质素降解酶系统的组成、分子生物学研究和主要影响因子的研究进展,以及国内外在外生菌根真菌产木质素降解酶方面的研究,阐述了其在有机污染土壤修复中的优势。木质素降解酶系统能够降解多环芳烃、氯代芳烃、硝基有机物和染料等多种环境污染物。许多外生菌根真菌都具有木质素降解酶系统。     

Cloning, expression in Escherichia coli, and enzymatic properties of laccase from Aeromonas hydrophila WL-11

A strain WL-11 with high laccase activity was isolated from activated sludge collected from the effluent treatment plant of a textile and dyeing industry.It was identified as Aeromonas hydrophila by physiological test and 16S rDNA sequence analysis.A gene encoding of laccase from a newly isolated Aeromonas hydrophila WL-11 was cloned and characterized.Nucleotide sequence analysis showed an open reading frame of 1605 bp encoding a polypeptide comprised of 534 amino acids.The primary structure of the enzyme predicted the structural features characteristic of other laccases,including the conserved regions of four histidine-rich copper-binding sites.The predicted amino acid sequence showed a high homology(more than 60%) with bacterial laccases in the genome and protein databases and the highest degree of similarity(61% identity) was observed with the multicopper oxidase of Klebsiella sp.601.When expressed in Escherichia coli,the recombinant enzyme was overproduced in the cytoplasm as soluble and active form.The purified enzyme had an optimum pH of 2.6 and 8.0 for ABTS(2,2'-azino-bis(3-ethylbenzthiazolinesulfonic acid) and DMP(2,6-dimethoxyphenol),respectively.The kinetic study on ABTS revealed a higher affinity of this enzyme to this substrate than DMP.     

Effects of nonionic surfactant Triton X-100 on the laccase-catalyzed conversion of bisphenol A

The laccase-catalyzed conversion of bisphenol A (BPA) in aqueous solutions was studied in the absence and presence of nonionic surfactant Triton X-100. It was found that the addition of Triton X-100 into the reaction system increased the conversion of BPA, especially near the critical micelle concentration of Triton X-100. Also it was found that the stability of laccase was greatly improved in the presence of TritonX-100. Studies on the endogenous fluorescence emission of laccase indicated that there existed an interaction between Triton X-100 and laccase, which was beneficial to folding and stabilizating of laccase. The binding of Triton X-100 to the laccase surface also mitigated the inactivation e ect caused by the free radicals and polymerization products. Under otherwise identical conditions, a lower dosage of laccase was needed for the higher conversion of BPA in the presence of Triton X-100.     

基因工程菌漆酶对蒽醌染料的脱色研究

利用基因工程菌甲醇毕赤酵母(PMAD16/pMETαA-Lcc1)培养产生的重组漆酶,研究了重组漆酶对蒽醌染料Remazol Brilliant Blue R脱色的影响。结果表明:重组漆酶对蒽醌染料RBBR脱色的最佳pH值为4.5,温度为45℃,当溶液中漆酶活力为10.0U/mL时,在最适脱色条件下作用14h,粗漆酶和纯化漆酶对蒽醌染料RBBR(80mg/L)的脱色率分别为95.2%和87.5%,重组漆酶可有效地使蒽醌染料RBBR脱色。     

生物表面活性剂逆胶束中漆酶催化性能的研究

对一种新型生物表面活性剂逆胶束中漆酶的催化性能进行了研究.以鼠李糖脂(RL,阴离子生物表面活性剂)为表面活性剂,异辛烷和正己醇为溶剂和助表面活性剂,构建逆胶束,分别考察了有机相组成、RL浓度、逆胶束体系含水率W0、缓冲溶液酸碱度及盐浓度等因素对该逆胶束体系中漆酶催化性能的影响.结果表明,当逆胶束有机相正己醇/异辛烷体积比为1:1.8,RL浓度为13mmol/L,体系含水率W0为42,缓冲溶液pH5.0,KCl浓度为10mmol/L时,该体系中漆酶具有较好的活性及稳定性.对比试验显示,与化学表面活性剂AOT逆胶束相比,RL逆胶束能使漆酶表现出较高的活性及稳定性,说明与传统化学表面活性剂相比生物表面活性剂在构建逆胶束酶体系上的具有一定的优越性.     

复配逆胶束体系中漆酶催化性能研究

在由鼠李糖脂和Tween-80构建而成的复配逆胶束媒介研究了漆酶的催化性能.采用紫外法考察了Tween-80摩尔含量、逆胶束含水量、水相酸碱度、水相盐度以及助表面活性剂种类等反应条件对漆酶催化性能的影响.研究结果表明,反应的最佳条件:Tween-80的摩尔含量为30%,体系含水量为20,缓冲溶液pH为4.5,KCI浓度为70mmol/L,助表面活性剂正己醇.并且在最佳条件下,漆酶在复配逆胶束体系中的酶活比在单相鼠李糖脂逆胶束中高1.44倍,比在水溶液中高4.35倍,研究结果表明复配表面活性剂在构建胶束酶学上具有很大的潜力.