菌株Bacillus sp.CLb芽孢漆酶对染料、模拟染料废水脱色的研究

[目的]研究芽孢杆菌菌株CLb的芽孢漆酶在介体的互作下对不同类型的染料及模拟染料废水的脱色效果。[方法]制备芽孢粗酶液,在介体的互作下,测定具有漆酶活性的粗酶液对活性亮蓝、活性黑、靛红和结晶紫的脱色效果,筛选出对染料脱色有促进作用的漆酶介体,并且研究芽孢漆酶在不同pH条件下对染料和模拟染料废水脱色的影响。[结果]菌株CLb的芽孢漆酶在无介体参与下只能使活性亮蓝和结晶紫脱色。在介体乙酰丁香酮（ACE）的作用下,芽孢漆酶对4种染料的脱色率均超过70%。在pH 7.0条件下,芽孢漆酶浓度为44.74 U/L和介体ACE浓度为1 mmol/L时对模拟染料废水的脱色效果最好,6 h脱色率超过80%。在介体乙酰丁香酮存在,pH 7.0的条件下,菌株CLb的芽孢漆酶对合成染料脱色效率高于pH 9.0时的脱色率。[结论]在介体乙酰丁香酮的存在下,芽孢漆酶对染料和模拟染料废水均具有较好的脱色效果。     

漆树酶的固定化——Ⅰ聚丙烯酰胺包埋法固定化漆树酶

漆树酶(BC.1.10.3.2)从生漆中提取后,用聚丙烯酰胺包埋固定化。研究了固定化的条件,以及固定化漆树酶的重复使用性和耐热性,测定了它的米氏常数,并与天然漆树酶进行了比较。固定化漆树酶的活力回收可达30％;重复使用十多次后其活性还保留约80％;在干燥状态下,固定化漆树酶非常耐热。     

Lignin-degrading activity of edible mushroom <Emphasis Type="Italic">Strobilurus ohshimae</Emphasis> that forms fruiting bodies on buried sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) twigs

Strobilurus ohshimae is an edible mushroom, and it specifically forms its fruiting bodies on buried sugi (Cryptomeria japonica) twigs. In this research, we studied lignindegrading activity of S. ohshimae. We isolated 18 strains of S. ohshimae from various regions of Japan, and determined their lignin degradation rates on sugi wood meal medium. All the strains of S. ohshimae degraded approximately 6%–12% of sugi lignin in 30 days, and these lignin degradation rates were 1.5–3 times higher than those of Trametes versicolor, which is a typical lignin-degrading fungus. Among the three main lignin-degrading enzymes, activity of lignin peroxidase and manganese peroxidase was not observed, while 4340U/g of laccase was produced in 30 days. To investigate the effect of wood species on lignin degradation by S. ohshimae, the lignin degradation rate and laccase productivity on sugi wood meal medium were compared with those on beech (Fagus crenata). In T. versicolor, both lignin degradation rate and laccase productivity were higher on beech than on sugi. Conversely, in S. ohshimae, lignin degradation rate and laccase productivity were higher on sugi than on beech. Therefore, it was suggested that coniferous lignin is not always difficult to degrade for the fungi that inhabit softwood. Part of this article presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004     

陈生漆加漆酶制作精制漆

用不干的陈生漆制作精制漆的方法，即加漆酶的方法，制出的精制漆，其主要性能和用好生漆制出的精制漆基本一样，而且还有很大的灵活性，可根据不同用途，配以不同量的漆酶和其它添加剂来制作各种精制漆。     

灰葡萄孢菌胞外大分子分泌物的植物毒性及漆酶性质鉴定

采用透析的方法提取灰葡萄孢菌分泌的大于14 kD的胞外大分子物质,将其接种到番茄和菊花叶片组织,检测灰葡萄孢菌胞外大分子分泌物的植物毒性,并以鞣酸和RB亮蓝为指示剂鉴定灰葡萄孢菌胞外大分子分泌物的漆酶性质。结果显示:灰葡萄孢菌胞外大分子分泌物可导致叶片组织坏死,具有植物毒性;接种灰葡萄孢菌后第3天,鞣酸和RB亮蓝平板出现不同程度的显色反应,第6天时平板几乎完全显色,胞外大分子物质具有漆酶性质。     

Laccase and phosphatase activities of the dominant ectomycorrhizal types in a lowland oak forest

The year-round dynamics of laccase and acid phosphatase activities of ectomycorrhizas (EM) were monitored in an oak forest, with a microplate photometric method using individual EM root tips. Among the most frequent and abundant EM morphotypes, those of Lactarius quietus and Cortinarius anomalus showed a peak of laccase activity in spring, while those of Xerocomus chrysenteron displayed highest laccase activities in summer and autumn. In comparison, less seasonal differences were observed in EM acid phosphatase activity. This suggests that the different EM types within the EM community perform complementary functions depending on season.     

Effects of flax lignin addition on enzymatic oxidation of poly(ethylene adipate) urethanes

Lignin use in polymer industry has recently become very attractive from both economical and environmental reasons. In the specific case of blending, the addition of low lignin amounts was found to improve the properties and extends the application field of other natural or synthetic polymers, but the effects of oxidative enzymes on resulting blends are widely unknown. Thereby, the current study was carried out to determine some potential effects of small flax lignin concentrations (4.2 and 9.3 wt%) on the enzymatic degradation of a poly(ethylene adipate) urethane (PU). Thin cast films of PU and its lignin blends were incubated for 3 days at 30 °C with buffered solutions of fungal peroxidase and laccase extracted from Aspergillus sp., and compared with the untreated ones. Changes in surface structure and morphology were investigated by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) and scanning electron microscopy (SEM), while the impact on bulk was assessed from tensile tests and thermogravimetry analysis (TGA). Although the addition of flax lignin reduces the surface structural modifications after enzyme treatment, the morphology, tensile and thermo-oxidative characteristics are still affected, with laccase showing the higher degradative efficiency. The lignin concentration, its high impact on the resulting blends morphology and relative low resistance to laccase and peroxidase degradation was the most important factors proved to driven the enzymatic oxidation.     

高寒草地土壤产漆酶真菌的筛选及分子鉴定

以高寒草地土壤分离出的9株真菌菌株为研究对象,采用rDNA-ITS方法对筛选出的产漆酶菌株进行分子鉴定,根据菌株在愈创木酚、蒽酮、邻苯二酚、邻联甲苯胺、邻甲苯胺、α-萘酚、联苯胺、没食子酸为底物的选择性培养基上的生长情况、菌落大小、漆酶催化氧化还原反应产生的变色圈直径大小及其颜色深浅程度,进行产漆酶真菌初筛,采用液体产酶发酵法选用ABTS为底物测定漆酶活力进行复筛。鉴定结果为菌株1.9为Uncultured fungus clone.,2.1a为Scytalidium,310b为Marasmius tricolor,2.1c为Saccharicola,2.3a为fungal sp,2.4d为Saccharicola.,10a为Marasmiusrotalis,3.7c为Alternaria.WB为Verticilliumlongisporum。筛选结果表明：除菌株3.7c以外,其余8株菌株在以愈创木酚、蒽酮、邻联甲苯胺、联苯胺为底物的培养基上基本都产生氧化带,在没食子酸底物培养基上均不产生氧化带;菌株1.9,2.1a,310b,2.4d,10a,2.1c,2.3a均具有产漆酶活力,其中菌株310b具有较强产漆酶活力。     

产酶微生物及其应用

介绍了产生纤维素酶和漆酶的真菌和细菌种类,以及各种酶在制浆造纸、饲料工业、污水治理、纺织洗涤、食品和堆肥等领域的应用情况。     

姬松茸4个品种菌丝体的生物酶活性测定

对本实验室保藏的4个姬松茸品种进行了菌丝发酵,以酪蛋白、PNPP和ABTS为底物测定了菌丝体细胞内的蛋白酶、磷酸酶和漆酶的活性。数据结果显示,被测定的4个姬松茸品种均具有磷酸酶的活性,其中姬松茸(巴西)的活性最高为48.1 U·g-1;蛋白酶和漆酶活性的检测中发现,只有姬松茸7号和姬松茸11号具有活性,其中姬松茸7号具有较高的蛋白酶活性(95.2 U·g-)1,姬松茸11号具有较高的漆酶活性(58.6 U·g-)1。姬松茸7号和姬松茸11号同时具有3种酶的活性,可以作为深加工的优势材料。