共查询到20条相似文献,搜索用时 109 毫秒
1.
《菌物学报》2017,(9):1243-1250
漆酶是一种含铜的多酚氧化酶,其作用底物范围广,在食品、能源和环保等领域具有重要的应用价值,因此筛选高产漆酶菌株对于其工业应用具有重要意义。本实验以78株香菇菌株为实验材料,利用愈创木酚平板法初筛,液体发酵复筛的方法筛选高产漆酶香菇菌株,并通过实时定量PCR分析香菇漆酶同工酶在不同菌株之间转录表达的差异性和特异性。结果表明,不同香菇菌株的生长速度、变色圈大小存在差异,以菌丝圈和变色圈大小及其比例为依据,从78株香菇菌株中筛选出19株潜在的漆酶高产菌株;液体发酵从这19株菌株中复筛出3株高产漆酶的菌株:香240、15和冕宁浅灰,其中香240在培养第3天酶活达到最高(3.583U/m L),香15和冕宁浅灰则在培养第7天酶活达到最高(分别为3.842U/m L和2.806U/m L);实时荧光定量PCR结果显示,10个漆酶同工酶基因在两株香菇菌株中的表达存在差异性和特异性,其中Lelac5和Lelac9只在菌株香240中检测到转录本;Lelac1–4在菌株15中表达量较高,而Lelac8、Lelac10和Lelac11在香240中的表达量较高。研究结果将为香菇漆酶的进一步研究提供基础。 相似文献
2.
真菌漆酶异源表达研究进展 总被引:3,自引:0,他引:3
由于漆酶能够氧化芳香类化合物和其它一些非芳香类有机物,具有广泛的底物特异性,因此在纸浆漂白、纺织品染料脱色、有毒废弃物的去除、生物修复和生物传感器等方面具有巨大的应用潜力。但是缺少大量廉价的酶源供应阻碍了漆酶商业化的应用,解决这个问题的一个主要方法就是通过漆酶的异源表达来获得大量的漆酶。综述了真菌漆酶在酵母表达系统和丝状真菌表达系统中表达的研究结果,着重总结了影响漆酶异源表达的因素和提高漆酶表达的策略。 相似文献
3.
从茶树内生真菌筛选产漆酶的菌株,分析不同营养因素和培养条件对菌株漆酶酶活力的影响。采用6种显色底物的平板初筛和酶活测定的复筛方法,从15株茶树内生真菌菌株中筛选获得1株产漆酶酶活较高的菌株CSN 4。单因素分析结果显示,液态发酵条件下菌株CSN-4适宜的主要培养基成分是麸皮和蛋白胨;菌株CSN-4分别在麸皮30 g/L、蛋白胨2.5 g/L、CuSO4·5H2O 0.015 g/L和茶水6 g/L时发酵产漆酶酶活最高。发酵条件试验结果表明,菌株CSN-4分别在接种量为6个菌饼(直径6 mm)、装液量60 mL/250 mL、pH 4.8、摇床转速120 r/min,培养温度为28 ℃时产漆酶酶活较高。在培养基中添加麸皮和茶水对菌株CSN-4产漆酶有明显的促进作用。经过培养基成分及培养条件优化后,菌株CSN 4产漆酶酶活显著升高,达到2 417 U/L。 相似文献
4.
5.
Trametes sp.AH28-2漆酶A的诱导合成及其基因5''''-端调控区的克隆与分析 总被引:5,自引:0,他引:5
Trametes sp.AH28—2漆酶同工酶的合成需要铜离子的存在,较高浓度的Cu^2 有利于漆酶合成。在以葡萄糖为碳源补加0.5mmol/L Cu^2 的培养基中生长时,发酵液漆酶活性为44.3u/L,同时补加4.0mmol/L邻甲苯胺时,漆酶酶活提高到71.0u/L;而在补加Cu^2 和邻甲苯胺的纤维二糖培养基中,酶活上升至2584u/L,为葡萄糖培养基的36.4倍。邻甲苯胺和铜离子诱导产生的漆酶同工酶组分,均为漆酶A(LacA)。竞争性RT—PCR分析表明,漆酶A基因(lacA)转录本的累积伴随有发酵液漆酶活性的增加,邻甲苯胺对lacA的调控发生在转录水平。lacA结构基因长2110bp,含有10个内含子;lacA的cDNA序列为1560bp,编码520aa的漆酶蛋白,其氨基酸序列与其它真菌漆酶具有较高的相似性。采用改进的反向PCR技术,扩增得到的lacA 5’-端调控区长1881bp,分析表明,该区域上分布有1个TATA框、7个CAAT框和多个潜在的顺式作用元件序列位点,包括5个MRE元件、9个CreA结合位点、4个XRE元件、2个STRE元件和7个氮因子调控位点等。这些序列位点的存在部分地对应了菌株摇瓶发酵奈件下lacA的表达规律。 相似文献
6.
利用聚丙烯酰胺凝胶等电聚焦圆盘电泳,分析比较了黑木耳(Auricularia auricula) 16个菌株和光帽鳞伞(Pfioliota nameko) 9个菌株的胞外漆酶同工酶酶谱。经多批次重复, 得到了所有菌株的恒定酶谱。依照这些酶谱及各酶带等电点(pT)值的不同,确立了它们的漆酶标志位点相似系谱。16株黑木耳被区分成4种类型9个组。9株光帽鳞伞被分成两种类 型3个组,但各组内之菌株间仍有异同。试验结果表明,把漆酶同工酶谱应用于菌株鉴别将是一种有意义的方法。 相似文献
7.
【目的】从26株真菌菌株中筛选高产漆酶菌株。【方法】采用愈创木酚法进行产漆酶菌株的筛选,通过正交实验对筛选出的高产菌株进行优化,并通过形态学和分子系统学对菌株进行鉴定。【结果】26株真菌菌株中有4株可产生漆酶,其中菌株H52.1为产漆酶最好菌株;菌株H52.1产漆酶优化培养基碳源为可溶性淀粉,氮源为硝酸铵,pH为8,金属离子为Ca2+;经鉴定,该菌株为大孢戴氏霉。【结论】大孢戴氏霉在产漆酶方面值得进一步研究开发。 相似文献
8.
大戟科4种药用植物内生真菌3种胞外酶活性的比较 总被引:8,自引:0,他引:8
利用分光光度计比色测定大戟科乌柏(Sapium sebiferum Roxb.)、大戟(Euphorbia pekinensis Rupr.)、泽漆(Euphorbia helioscopia Level)、重阳木(Bischofia polycarpam Airy Shaw)4种药用植物新鲜叶和茎叶内生真菌的纤维素酶、漆酶和多酚氧化酶活性,并比较它们的差异。结果表明:茎内的真菌漆酶活性和多酚氧化酶活性普遍比叶内真菌高;重阳木内生真菌的漆酶及多酚氧化酶活性比乌柏、大戟和泽漆的内生真菌高。不同植物以及同一植物不同部位的内生真菌的胞外酶活性存在较大差异,推测其机理可能与内生真菌的种类和宿主植物之间的相互关系等因素有关。从乌柏和重阳木茎内皮中筛选到的漆酶和多酚氧化酶活性较高的2个菌株有良好的应用前景。 相似文献
9.
利用聚丙烯酰胺凝胶等电聚焦圆盘电泳,分析比较了黑木耳(Auricularia auricula)16个菌株和光帽鳞伞(Pholiota nameko)9个菌株的胞外漆酶同工酶酶谱。经多批次重复,得到了所有菌株的恒定酶谱。依照这些酶谱及各酶带等电点(pI)值的不同,确立了它们的漆酶标志位点相似系谱。16株黑木耳被区分成4种类型9个组。9株光帽鳞伞被分成两种类型3个组,但各组内之菌株间仍有异同。试验结果表明,把漆酶同工酶谱应用于菌株鉴别将是一种有意义的方法。 相似文献
10.
依照真菌漆酶催化特定作用底物进行氧化还原反应结果的特性,设计了一个采用愈创木酚和α-萘酚作为酶反应显色判别依据和产酶菌株筛选的限制性驯化剂,同时结合选用木质素作为产酶诱导强化因子的筛选分析模版,对保藏和外购的常见大型担子菌和食用真菌进行产酶性能分析。结果发现,这些菌株均不能有效地生产漆酶或酶活性很低,以该模版作为初筛手段从杨树林等野生菌菇生长地中分离得到15株产酶野生微生物,经反复筛选得到3株酶活性较高的大型真菌株A21、X18和Y11,其胞外漆酶活力分别为255.9、248.7 和277.3 U·ml-1,具有可观的产业化生产开发应用潜力。 相似文献
11.
12.
During the last decade the search for novel biotechnologically valuable laccases/oxidases with a high redox potential and concomitant activity under neutral-alkaline conditions is an attractive and at the same time complicated task due to their rare occurrence in nature. By means of the modified micromethod based on the chromogenic reaction with indicator substrates the successful identification of laccases/oxidases with different pH-optimum was carried out during submerged cultivation of 71 fungal strains of different taxonomic groups. Based on more sensitivity (detected laccase activity can be 4–6 time less as compared with the usual spectrophotometric assay of laccase activity), good productivity (measurements of numerous samples at once in small total volume – up to 150 μL), economy and rapidity, the presented modification of chromogenic reaction can be applied for identification of trace amount of laccase/oxidase activity in biological liquids, to determine the chemoselectivity of induced laccase/oxidase isoforms with respect to pH-value of medium, and to monitor the dynamics of expression of alkaliphilic and acidophilic laccases/oxidases during submerged cultivation of fungi. 相似文献
13.
14.
15.
We investigated the influence of potential laccase inducers with environmental relevance on extracellular laccase activity and removal of the xenoestrogen technical nonylphenol (tNP) by the aquatic hyphomycete Clavariopsis aquatica. Concomitantly, we identified two putative laccase gene fragments (Icc1 and Icc2) and have followed their expression during removal of tNP under different conditions. Our results indicate a significant effect of copper on extracellular laccase activity in supernatants of fungal cultures. Laccase activity was highest in the presence of copper when added together with vanillic acid, followed by copper when used alone. Only slight laccase activities were recorded in the presence of only vanillic acid, whereas in the absence of either compound laccase activities were negligible. Laccase activity was well correlated with the removal efficiency of tNP, indicating the involvement of laccase in tNP bioconversion. Overall, Icc2 was less expressed than Icc1. The expression of Icc1 and Icc2 correlated only partially with the measured laccase activity, suggesting the existence of cell-associated laccase fractions not detectable in fungal culture supernatants and/or the existence of additional laccase genes. 相似文献
16.
17.
18.
A comparative study has been conducted on seven white rot fungi to investigate their abilities to produce laccase and selectively degrade lignin. Laccase was produced constitutively on the different media tested. Of the different lignins, phenolic compounds and sugars involved, the highest laccase yield was obtained on indulin AT. Salicylic acid inhibited enzyme activity. A temperature of 20°C and 0.2% of indulin AT were found to be optimum for enzyme activity. No correlation was found between the amount of enzyme and fungal mass produced. During semisolid degradation of angiospermic wood sawdust, Daedalea flavida caused a total weight loss of 11%, with a lignin loss of 15.77% during two months of decay. Lignin removal was comparatively selective during the first month, during which time laccase production was also higher, indicating its probable role in lignin degradation. 相似文献
19.