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一株产漆酶菌株的筛选鉴定和发酵条件的研究 总被引:2,自引:0,他引:2
以愈创木酚为底物,采用平板筛选法筛选得到一株产漆酶菌株WS1-2,形态学特性表明该茵属于绿色木霉.对产酶条件的初步研究结果表明,WS1-2菌株的产酶高峰期出现在接种培养后的第4d.与蔗糖、乳糖、半乳糖和可溶性淀粉相比,以葡萄糖为碳源时,发酵上清液的漆酶活力明显要高,最大值达230U/L.以NH4Cl为氮源,最有利于WS1-2漆酶的产生,漆酶活力最高可达到234U/L.0.01mmol/L的愈创木酚和ABTS对WS1-2产漆酶有明显的诱导作用,3~5mg/L的Tween-80可以明显提高WS1-2的产酶水平. 相似文献
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本研究经过选择性培养基分离及特异性底物筛选从干旱区荒漠植物松叶猪毛菜和红砂灌丛土壤中分离出两株产漆酶的真菌Z45和H53,扩增其ITS序列进行分子鉴定,并分析温度、碳源、氮源、碳氮比和pH对菌株Z45生长的影响,在此基础上利用正交试验优化其培养条件.结果 表明,产漆酶真菌Z45和H53均隶属于端梗霉属(Acrophialophora sp.),Z45温度适应范围广且耐高温,最适生长温度为40℃.最优培养基组合为:分别以麦芽糖和硝酸铵为碳、氮源,按15∶1的碳源/氮源比配制,pH值6.0.本研究从干旱区荒漠土壤中获得了1株耐高温、产漆酶、应用价值高的子囊菌端梗霉,丰富了产漆酶微生物的种质资源库. 相似文献
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高酶活菌株的筛选及漆酶特性 总被引:5,自引:0,他引:5
通过Bavendamn氏反应和液体发酵实验筛选出漆酶高产菌株 ,并对其产酶条件和酶活性进行了研究。结果表明 71株实验真菌中有 64株Bavendamn氏反应呈阳性 ,且阳性菌株都具有漆酶活性 ;不同菌株产酶培养基最适碳源、氮源不同 ,采绒革盖菌以淀粉为碳源、干酪素为氮源 ,毛栓菌以麦草粉为碳源、硫酸铵为氮源 ,有利于酶的分泌 ;不同来源漆酶性质不尽相同 ,采绒革盖菌漆酶最适酶解温度为 2 5℃ ,最适酶解pH值为4.6,毛栓菌则分别为 3 0℃和 pH 4.0 ;K+ ,Zn2 + 等对 2种漆酶均有激活作用 ,Ag+ 则能明显抑制漆酶活性。 相似文献
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Masayoshi Takakuwa Yasuo Watanabe 《Bioscience, biotechnology, and biochemistry》2013,77(10):2167-2173
Variations in lipid components of washings and homogenate of pressed baker’s yeast were investigated during the storage of pressed baker’s yeast at 30°C. Washings represents the substances which had leaked out from cells. Homogenate represents those contained in whole cells. Lipids in yeast washings increased toward softening, the phospholipids in yeast homogenate decreased continuously during storage. Two stages, an earlier period of storage (Stage I) and a later period of storage (Stage II) were observed in the degradation of phospholipids. Free fatty acid which was the main degradation product of phospholipid accumulated in Stage II, particularly at softening. The order in phospholipid degradation was PC>PE>PI + PS (PI>PS). Moreover, when washings of stored yeast at softening were assayed using 14C-acyl PC, the release of 14C-acyl fatty acid was observed.These results suggest that phospholipids were degraded by some phospholipid-deacylating enzymes toward softening. From the results of lipid analysis, we inferred that the responsible enzymes were phospholipases. 相似文献
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Oxidation of carbazole, N-ethylcarbazole, fluorene, and dibenzothiophene by the laccase of Coriolopsis gallica 总被引:3,自引:0,他引:3
David C. Bressler Phillip M. Fedorak Michael A. Pickard 《Biotechnology letters》2000,22(14):1119-1125
Purified laccase from Coriolopsis gallica UAMH8260 oxidized carbazole, N-ethylcarbazole, fluorene, and dibenzothiophene in the presence of 1-hydroxybenzotriazole and 2,2-azinobis (3-ethylbenzthiazoline-6-sulfonic acid) as free radical mediators. Susceptibility to laccase oxidation appears related to the ionization potential (IP) of the substrate: compounds with an IP above 8.52, dibenzofuran (IP = 8.77) and benzothiophene (IP = 8.73) were not attacked. Carbazole (IP = 7.68) was the most sensitive to oxidation with >99% transformed with 10 milliunits of laccase after 1 h, though most reactions were carried out for 18 h. 9-Fluorenone was identified as the product of fluorene (IP = 8.52) oxidation, and dibenzothiophene sulfone from dibenzothiophene (IP = 8.44). Although carbazole and N-ethylcarbazole were both completely removed within 18 h, no oxidation or condensation metabolites were detected. This investigation is the first to report the oxidation of dibenzothiophene, carbazole, and N-ethylcarbazole by laccase. 相似文献
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漆酶是一种含铜的多酚氧化酶,与植物病原菌致病性、黑色素合成及降解木质素等方面相关。为明确漆酶在新月旋孢腔菌的催化作用及其催化活性,以2,2′-连氮-双(3-乙基苯并噻唑-6-磺酸)(简称ABTS)为底物,利用分光光度计在420nm下测定胞内漆酶活力,结果表明酶活测定最佳反应条件为缓冲液pH2.8、Cu2+浓度500μmol/L和0.6mmol/L ABTS。根据漆酶Cu2+结合保守结构域设计了1条引物,对新月旋孢腔菌漆酶基因进行克隆,并通过RACE技术克隆了其全长cDNA序列。开放阅读框长1,803bp, 相似文献
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漆酶是一种含铜的多酚氧化酶[1],广泛分布于植物、真菌、少数昆虫和细菌中,可用于纸浆造纸、生物合成、食品、能源、木材加工、环保、改善纤维特性、生物检测等多个领域。鉴于其重要的应用价值,漆酶正日益受到重视。 相似文献
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Brandt Bertrand Fernando Martínez‐Morales María R. Trejo‐Hernández 《Biotechnology progress》2017,33(4):1015-1034
Improving laccases continues to be crucial in novel biotechnological developments and industrial applications, where they are concerned. This review breaks down and explores the potential of the strategies (conventional and modern) that can be used for laccase enhancement (increased production and upgraded biochemical properties such as stability and catalytic efficiency). The challenges faced with these approaches are briefly discussed. We also shed light on how these strategies merge and give rise to new options and advances in this field of work. Additionally, this article seeks to serve as a guide for students and academic researchers interested in laccases. This document not only gives basic information on laccases, but also provides updated information on the state of the art of various technologies that are used in this line of investigation. It also gives the readers an idea of the areas extensively studied and the areas where there is still much left to be done. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1015–1034, 2017 相似文献
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《Biocatalysis and Biotransformation》2013,31(1):86-95
Conventional cross-linking of proteins involves the use of toxic chemicals. Here, cross-linking of gelatine and gelatine hydrolysates with tyrosinases from Botryosphaeria obtusa (BoT1 and BoT2), Agaricus bisporus (AbT) and from Verrucomicrobium spinosum (VsT) and with laccases from Trametes hirsuta (ThL) and T. versicolor (TvL) was demonstrated. Enzymatic oxidation of tyrosine residues was indicated by UV/VIS and fluorescence spectroscopy and further confirmed by oxygen consumption measurements. Using a model substrate (Tyr-Ala) dimerization was demonstrated by using RP-HPLC and LC-MS. Enzymatic cross-linking significantly increased the molecular weight of the soluble material up to the point of precipitation as demonstrated by both SDS-PAGE and size exclusion chromatography. The effect of cross-linking was further enhanced in the presence of phenolic molecules such as catechin. 相似文献
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The use of insertional mutagenesis to discover genes that impact laccase activity has resulted in the identification of multiple cellular processes that affect the fitness of Cryptococcus neoformans. Fitness has been defined as the ability of an organism to propagate and evolve within a given environment. Because the human host is an evolutionary dead-end for an opportunistic pathogen, we have defined pathogenic fitness here as the capability to successfully propagate within the stressful environment of the host, causing disease by expression of virulence traits that damage the host. In this review, laccase-deficient insertional mutants will be highlighted in terms of the basic biological processes in which they are involved. The impact of laccase-associated cellular functions on fitness and virulence will be discussed, as will the mutants' potential as therapeutic targets. Vacuolar function, copper homeostasis, mitochondrial function and carbon repression are covered. 相似文献
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Marc Dubernet Pascal Ribereau-Gayon Henri R. Lerner Eitan Harel Alfred M. Mayer 《Phytochemistry》1977,16(2):191-193
The partial purification of an extracellular laccase from Botrytis cinerea is described. Specificity of the enzyme, its Km for a number of substrates and sensitivity to some inhibitors are described. The enzyme is a typical laccase but has an exceptionally low pI and great stability to acid pH. On gel electrophoresis two isoenzymes could be detected. 相似文献