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Effects of copper ions on the laccase activities and gene transcription level of Auricularia heimuer
MI Haoyu, GENG Nannan, DU Pengyu, LIU Zengcai, ZOU Li
Mycosystema ›› 2025, Vol. 44 ›› Issue (4) : 240271.
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Effects of copper ions on the laccase activities and gene transcription level of Auricularia heimuer
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The effect of copper ions on laccase activity and transcriptional expression of laccase genes in Auricularia heimuer “Hei Hou Yuan” is investigated. The results show that as the copper ion concentration increases from 100 to 700 μmol/L, extracellular laccase activity in A. heimuer increases significantly. Copper ion concentration strongly influences the transcriptional levels of laccase genes. Specifically, the transcription levels of AhLac1, AhLac3, and AhLac6 are significantly upregulated as the copper ion concentration rises. The expression levels of AhLac2, AhLac4, and AhLac5 peak at 500 μmol/L, and increase by 3.38-, 1.29-, and 1.59-fold, respectively. Under different induction times, the transcription levels of AhLac1, AhLac3, and AhLac6 show varied degrees of upregulation with the extension of induction period. Correlation analysis further reveals a positive relationship between laccase activity and the transcriptional levels of AhLac1, AhLac3, and AhLac6. This study provides a scientific basis for improving the yield of A. heimuer and investigating the molecular regulatory mechanisms underlying laccase gene expression.
Auricularia heimuer / copper ion / laccase activity / transcription level {{custom_keyword}} /
Table 1 Content of main active components in CME表1 CME中主要活性成分含量 |
| 成分 Components | 含量 Content (mg/g) |
|---|---|
| 多糖 Polysaccharide | 509.47±14.73 |
| 尿苷Uridine | 1.21±0.10 |
| 鸟苷Guanosine | 0.66±0.03 |
| 腺苷Adenosine | 1.47±0.11 |
| 虫草素 Cordycepin | 2.66±0.15 |
| N6-(2-羟乙基)腺苷 N6-(2-hydroxyethtl)-adenosine | 2.15±0.09 |
| 总多酚 Total polyphenols | 9.21±0.37 |
| 类胡萝卜素 Carotenoid | 0.95±0.04 |
Fig. 1 Determination of monosaccharide composition by HPLC. A: Mixed reference substance; B: Tested samples. 1: Man; 2: Rha; 3: Glc-UA; 4: Gal-UA; 5: Lactose (internal standard); 6: Glc; 7: Gal: 8: Xyl; 9: Ara; 10: Fuc.图1 HPLC检测单糖组成 A：混合对照品；B：供试品. 1：甘露糖；2：鼠李糖；3：葡萄糖醛酸；4：半乳糖醛酸；5：乳糖内标；6：葡萄糖；7：半乳糖；8：木糖；9：阿拉伯糖；10：岩藻糖 |
Table 2 Monosaccharide composition of Cordyceps militaris polysaccharide表2 蛹虫草多糖的单糖组成 |
| 样品 Sample | 甘露糖 Man | 半乳糖醛酸 Gal-UA | 葡萄糖 Glc | 半乳糖 Gal | 阿拉伯糖 Ara |
|---|---|---|---|---|---|
| 含量 Content (mol%) | 25.73 | 9.26 | 29.01 | 29.39 | 5.62 |
Fig. 2 Determination of nucleosides by HPLC. A: Mixed reference substance; B: Test sample. 1: Cytidine; 2: Uridine; 3: Guanosine; 4: Thymidine; 5: Adenosine; 6: Cordycepin; 7: N6-(2-hydroxyethtl)- adenosine.图2 HPLC检测核苷类成分 A：混合对照品；B：供试品. 1：胞苷；2：尿苷；3：鸟苷；4：胸苷；5：腺苷；6：虫草素；7：N6-(2-羟乙基)腺苷 |
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| [2] |
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| [3] |
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| [5] |
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| [6] |
The expression of laccase in the white rot fungus Trametes versicolor is regulated at the level of gene transcription by copper and nitrogen. We used reverse transcription-PCR to demonstrate that as the concentration of copper or nitrogen in fungal cultures was increased, an increase in laccase activity, corresponding to increased laccase gene transcription levels, was observed. In addition, we demonstrated that the amounts of laccase mRNA and laccase activity in 10-day-old cultures were a direct function of the concentration of either 1-hydroxybenzotriazole, a previously described laccase substrate, or 2,5-xylidine, a well-known laccase inducer, in the medium. No induction was observed after the addition of two aromatic acids, ferulic acid and veratric acid, which have been shown to induce laccase production in other white rot fungi. When either copper, 2,5-xylidine, or both compounds were added to cultures grown in the absence of copper, increased laccase transcript levels were detected within 15 min. Corresponding increases in laccase activity were observed after 24-h incubation only when copper was present.
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| [7] |
More and more people have recently payed their attention to studies or investigations on medicinal fungi in China. However, many nomenclatural inconsistencies were found in the Chinese reports. The publications on Chinese medicinal fungi were critically reviewed. As a result, 473 medicinal fungi from China were enumerated in the present checklist. All of the names were checked or revised in accordance with the contemporary taxonomy and the latest version of the International Code of Botanical Nomenclature (Vienna Code). The ‘out-of-date’ names, ill-names, and un-existed names and misapplied names in the previous reports were revised. The common synonyms were listed after their valid names. The main medicinal functions of each species, together with the original or important references, were provided.
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| [8] |
Macrofungi, as an important component in forest ecosystems, consist of most members of Basidiomycota and some members of Ascomycota, having important economical value and ecological functions. Extensive field investigations have been carried out in almost whole types of the Chinese forests during the past 30 years, and 112 000 specimens were collected. Based on morphological examination and phylogenetic analyses in combination with ecology and biogeography, 4 250 species belonging to 21 orders in Baidiomycota and Ascomycota were identified, including two new families, four new subfamilies, 69 new genera and 885 new species. Yunnan Province is the richest in macrofungal diversity among provinces or regions in China, and 314 new species were described from this province, accounting for 35% of all the new species described from China by the authors. Our studies have made contributions to deepening the understanding of global diversity of macrofungi. The names of some important Chinese medicinal fungi were revised, the diversity characteristics of Chinese poisonous mushrooms were revealed, and the pathogenetic wood-decaying species were ascertained. These data improved our knowledge on utilization of natural resources and protection of forest health. Based on molecular evidences, the origin of some forest representative fungal genera or species complex were deduced, and their dispersal and speciation were discussed, for the purposes of providing some data for evolutionary study at level of family, order or class of macrofungi henceforth. {{custom_citation.content}}
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| [9] |
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| [10] |
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| [11] |
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| [12] |
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| [13] |
Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/μg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus. Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent.
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| [14] |
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| [15] |
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| [16] |
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| [17] |
Gaeumannomyces graminis var. tritici, a filamentous ascomycete, is an important root pathogen of cereals that causes take-all disease and results in severe crop losses worldwide. Previously we identified a polyphenol oxidase (laccase) secreted by the fungus when induced with copper. Here we report cloning and partial characterization of three laccase genes (LAC1, LAC2, and LAC3) from G. graminis var. tritici. Predicted polypeptides encoded by these genes had 38 to 42% amino acid sequence identity and had conserved copper-binding sites characteristic of laccases. The sequence of the LAC2 predicted polypeptide matched the N-terminal sequence of the secreted laccase that we purified in earlier studies. We also characterized expression patterns of these genes by reverse transcription-PCR. LAC1 was transcribed constitutively, and transcription of LAC2 was Cu inducible. All three genes were transcribed in planta; however, transcription of LAC3 was observed only in planta or in the presence of host (wheat) plant homogenate.
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| [18] |
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| [19] |
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| [20] |
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| [21] |
Pleurotus ostreatus is a white rot basidiomycete that produces several extracellular laccase isoenzymes, including phenol oxidase A1b (POXA1b), POXA2, and POXC. POXC was the most abundant isoenzyme produced under all of the growth conditions examined in this study. Copper was the most efficient inducer of laccase activity among the putative inducers tested. The amounts of all of the previously described laccase isoenzymes increased substantially in copper-supplemented cultures. Under these conditions expression of POX isoenzymes was regulated at the level of gene transcription. It is worth noting that poxa1b mRNA was the most abundant induced transcript at all of the growth times analyzed, and the amount of this mRNA increased until day 7. The discrepancy between the poxa1b transcript and protein amounts can be explained by the presence of a high level of the protein in P. ostreatus cellular extract, which indicated that the POXA1b isoenzyme could be inefficiently secreted and/or that its physiological activity could occur inside the cell or on the cell wall. Moreover, the POXA1b isoenzyme behaved uniquely, as its activity was maximal on the second day of growth and then decreased. An analysis performed with protease inhibitors revealed that the loss of extracellular POXA1b activity could have been due to the presence of specific proteases secreted into the copper-containing culture medium that affected the extracellular POXA1b isoenzyme.
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| [22] |
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| [23] |
The white-rot fungus Pleurotus eryngii secretes various laccases involved in the degradation of a wide range of chemical compounds. Since the laccase production is relatively low in fungi, many efforts have been focused on finding ways to increase it, so in this study, we investigated the effect of copper on the transcription of the pel3 laccase gene and extracellular laccase activity. The results indicate that adding 0.5 to 2 mM copper to liquid cultures of P. eryngii KS004 increased both pel3 gene transcription and extracellular laccase activity in a concentration-dependent manner. The most significant increase in enzyme activity occurred at 1 mM Cu, where the peak activity was 4.6 times higher than in control flasks. Copper also induced the transcription of the laccase gene pel3. The addition of 1.5 and 2 mM Cu to fungal culture media elevated pel3 transcript levels to more than 13-fold, although the rate of induction slowed down at Cu concentrations higher than 1.5 mM. Our findings suggest that copper acts as an inducer in the regulation of laccase gene expression in P. eryngii KS004. Despite its inhibitory effect on fungal growth, supplementing cultures with copper can lead to an increased extracellular laccase production in P. eryngii.© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.
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| [24] |
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| [25] |
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{{custom_citation.annotation}}
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| [26] |
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{{custom_citation.annotation}}
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| [27] |
{{custom_citation.content}}
{{custom_citation.annotation}}
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| [28] |
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{{custom_citation.annotation}}
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| [29] |
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{{custom_citation.annotation}}
|
| [30] |
{{custom_citation.content}}
{{custom_citation.annotation}}
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| [31] |
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| [32] |
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| [33] |
Laccases are so called as green catalysts since they can reduce a vast of substrates including lignocellulose and other macromolecular polymers into water and low-molecular weight by-products, without formation of secondary pollutants. Therefore, laccases have huge potentials in various industrial applications. For free laccases, immobilization treatment is capable of enhancing their capacities to adapt harsh conditions for industrial utilization, thermostability, reusability, and expanding the tolerant ranges of pH and temperature. As fungal laccases are important origins of laccases, their properties, production, current immobilization strategies, and potential applications in various fields were summarized in this review. {{custom_citation.content}}
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| [34] |
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| [35] |
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{{custom_citation.annotation}}
|
| [36] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [37] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [38] |
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{{custom_citation.annotation}}
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| [39] |
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{{custom_citation.annotation}}
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| [40] |
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{{custom_citation.annotation}}
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| [41] |
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| [42] |
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{{custom_citation.annotation}}
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| [43] |
{{custom_citation.content}}
{{custom_citation.annotation}}
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| [44] |
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{{custom_citation.annotation}}
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| [45] |
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| [46] |
A novel laccase (Tolacc-T) from white rot fungus Trametes orientalis was enriched to apparent homogeneity with a specific activity of 20.667U/mg protein and recovery yield of 47.33%. The SDS-PAGE gave a single band indicating that Tolacc-T appears as a monomeric protein with a molecular mass of 44.0kDa. Domain structure analysis revealed that Tolacc-T contained a typical copper II binding domain and shared three potential N-glycosylation sites, but had no copper I binding domain, demonstrating that the enzyme is really a laccase, but a novel laccase. Optimal pH and temperature of Tolacc-T was 4.0 and 80°C, respectively, and it retained more than 80% of its original activity after 2h incubation at 10°C to 50°C. The enzyme exhibited strict substrate specificity towards ABTS but showed no or trace activities towards other substrates. Among the metals tested, Mn was proved to be the best activator for enhancing the laccase activity. A strongly inhibiting effect was found when NaN, -cysteine, and DTT were added to the enzyme. However, Tolacc-T activity was little bit inhibited in the presence of chelator EDTA. Furthermore, the enzyme was capable of degrading structurally different synthetic dyes in the absence of a redox mediator.Copyright © 2017 Elsevier B.V. All rights reserved.
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| [47] |
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| [48] |
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| [49] |
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| [50] |
安琪, 吴雪君, 吴冰, 戴玉成, 2015. 不同碳源和氮源对金针菇降解木质纤维素酶活性的影响. 菌物学报, 34(4): 761-771
以3株栽培的金针菇Flammulina velutipes为材料,研究它们在玉米芯和棉子壳以及不同碳源、氮源培养条件下纤维素、半纤维素和木质素降解酶活性的规律。结果表明,不同金针菇菌株的羧甲基纤维素酶、木聚糖酶和漆酶活力显著不同（P<0.001）,同时,培养条件对羧甲基纤维素酶、木聚糖酶和漆酶的活力都有显著影响（P<0.001）。在简单碳源存在的条件下,金针菇的羧甲基纤维素酶和木聚糖酶活力远远低于复杂碳源培养基（P<0.05）。全营养培养基上生长的金针菇的羧甲基纤维素酶和木聚糖酶活力低于缺乏碳源和氮源的培养基（P<0.05）。漆酶活力在无简单氮源培养基上低于全培养基（P<0.05）和无葡萄糖培养基（P<0.05）,即复杂碳源和氮源培养基上的漆酶活力低于简单碳源和氮源培养基（P<0.05）。
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| [51] |
曹可可, 刘宁, 马双新, 曹志艳, 梁东旭, 柴江婷, 董金皋, 2015. 大斑刚毛座腔菌高产漆酶条件的响应面优化及酶学特性. 中国农业科学, 48(11): 2165-2175
【目的】优化大斑刚毛座腔菌（Setosphaeria turcica）高产漆酶的最佳发酵条件，确定其酶学性质，为进一步开发应用奠定基础。【方法】以大斑刚毛座腔菌为出发菌株，首先采用单因素试验确定影响菌株产漆酶的碳源、氮源及铜离子的种类及范围，并在单因素试验的基础上，以漆酶酶活力为响应值，采用central composite design（CCD）响应面设计试验，利用Design Expert软件对响应值进行3因素3水平下的多元二次回归拟合分析，优化大斑刚毛座腔菌产漆酶的发酵培养基成分；初步分离大斑刚毛座腔菌发酵液中的漆酶，以ABTS为反应底物，设置不同温度及pH，测定漆酶的最适反应温度、pH及热稳定性、pH稳定性，进一步测定其反应动力学常数Km值、Vm值，确定其酶学特性。【结果】建立了以漆酶酶活力为响应值的多元二次回归模型，模型差异显著（P=0.0001），可以用该模型来拟合试验；响应面分析结果表明，各因素对漆酶活力的影响大小依次为Cu<sup>2+</sup>>葡萄糖>尿素，而葡萄糖和尿素交互作用极显著；通过拟合求出模型极值点，对应的大斑刚毛座腔菌产漆酶的最佳培养条件为：葡萄糖50.05 g?L<sup>-1</sup>，KH<sub>2</sub>PO<sub>4</sub> 1 g?L<sup>-1</sup>，尿素1.46 g?L<sup>-1</sup>，MgSO<sub>4</sub> 0.5 g?L<sup>-1</sup>，蛋白胨2 g?L<sup>-1</sup>，玉米浆0.5 g?L<sup>-1</sup>，CuSO<sub>4</sub> 0.07 g?L<sup>-1</sup>，Tween80 3 mL?L<sup>-1</sup>，28℃，150 r/min振荡培养7 d；在此条件下漆酶活力最高达（40.00±1.20）U?mL<sup>-1</sup>。对大斑刚毛座腔菌漆酶发酵液初步分离，经SDS-PAGE检测其漆酶相对分子量约为80 kD；以ABTS为底物时，最适反应温度为50℃，最适pH为4.2，在温度较高且弱酸性条件下活性较高，并且具有良好的温度稳定性和pH稳定性，常温下pH为4.2保持14 h后酶活力基本不变，50℃保温14 h后漆酶活力仍保持在60%以上；进一步在常温、pH为4.2时测定其米氏常数Km值为0.036 mmol?·L<sup>-1</sup>，最大反应速率Vm为28.63 mmol?L<sup>-1</sup>?min<sup>-1</sup>。【结论】利用大斑刚毛座腔菌液体发酵产漆酶并研究其酶学特性，表明作为玉米致病菌的大斑刚毛座腔菌产漆酶具有发酵周期短、活性高、稳定性好等特性，可进一步研究开发应用。
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| [52] |
曹永佳, 马鸿飞, 崔宝凯, 司静, 戴玉成, 2021. 不同固体发酵培养基下三种白腐真菌分泌的木质纤维素酶活性. 菌物学报, 40(5): 1123-1139
本研究选取众所周知的典型白腐真菌树舌灵芝Ganoderma applanatum、毛栓孔菌Trametes hirsuta和木蹄层孔菌Fomes fomentarius作为研究对象,对其利用木质纤维生物质进行发酵及添加有机营养、无机盐、金属离子、表面活性剂等进行了探索,期间以测定漆酶、滤纸纤维素酶、木聚糖酶活性表征3种菌株对木质纤维生物质的预处理能力,为确定白腐真菌菌株及单环境因子而达到高效预处理木质纤维生物质提高生物转化效率的目的奠定了重要的理论基础。结果显示,3种菌株分泌的木质纤维素酶在10周内基本都呈现先上升后下降的趋势,且酶活都较高,均可作为木质纤维生物质预处理的备选菌株。相比于针叶树（落叶松）基质,阔叶树（白桦）基质更适宜于3种菌株生长及分泌木质纤维素酶。各环境因子中,Cu<sup>2+</sup>的添加可提高漆酶活性,表面活性剂对于3种酶活的诱导作用均十分显著。
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{{custom_citation.annotation}}
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| [53] |
陈带娣, 牛杰振, 余晓媛, 严金平, 伊日布斯, 2013. 影响真菌漆酶表达及其活性的因素. 生命科学, 25(11): 1053-1058
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [54] |
戴玉成, 杨祝良, 2008. 中国药用真菌名录及部分名称的修订. 菌物学报, 27(6): 801-824
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [55] |
戴玉成, 杨祝良, 崔宝凯, 吴刚, 袁海生, 周丽伟, 何双辉, 葛再伟, 吴芳, 魏玉莲, 员瑗, 司静, 2021. 中国森林大型真菌重要类群多样性和系统学研究. 菌物学报, 40(4): 770-805
大型真菌主要为担子菌门的真菌和少数为子囊菌门的真菌,该类真菌具有重要的经济价值和生态功能,主要生长在森林生态系统中。30年来作者对我国几乎所有类型森林生态中的大型真菌进行了系统调查和采集,共采集标本11.2万号。基于对这些材料的形态学及分子系统学研究,并结合生态学和生物地理学特征,共鉴定出中国森林大型真菌4 250种,隶属于担子菌门和子囊菌门的21个目,发现和发表2个新科、4个新亚科、69个新属和885个新种。云南省是我国森林大型真菌最丰富的省份,描述于该省的新种有314种,占作者发表的全部中国新种的35%。这些研究为深入认识全球大型真菌物种多样性提供了中国的贡献,更新了我国重要食药用菌名称,揭示了我国毒蘑菇多样性基本特征,系统论述了我国森林病原菌的物种多样性,为资源利用、森林健康和保护提供了科学依据;论述了森林大型真菌代表性类群在种和属级水平的起源和演化,为今后开展重要类群科级、目级甚至纲级的系统进化关系提供了重要数据。
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [56] |
董玲玲, 谭晴, 张梦婷, 肖春丽, 张贺, 蒲金基, 刘晓妹, 2022. 外源物对杧果胶孢炭疽病菌漆酶活性及漆酶基因家族成员表达的作用. 果树学报, 39(7): 1157-1166
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [57] |
冯煊, 张小雪, 刘猛, 刘月涵, 张惠, 魏欢, 杨志新, 2020. Cu(Ⅱ)对一株镰刀菌形态发育及木质素酶系活性的影响. 菌物学报, 39(5): 839-847
本试验采用室内摇瓶培养,通过添加不同浓度的Cu(II),研究了Cu(II)对降解菌Fusarium sp. ZH-H2形态发育及木质素酶系活性的影响,以期诱导提高木质素酶系的活性。结果表明,当Cu(II)浓度为0.50mmol/L时,菌丝生物量最高达0.38g,菌丝团呈洁白平滑球状且数量最多,此浓度下木质素过氧化物酶（lignin peroxidase,LiP）活性峰值也达到最高,为28 315.41U/L,当Cu(II)浓度为2.00mmol/L时,有利于诱导锰过氧化物酶（manganese peroxidase,MnP）和漆酶（laccase,Lac）的代谢,最高峰值分别为1 612.90U/L、7 829.12U/L,并且明显缩短了峰值的形成时间,浓度过高或过低则对3种木质素酶系活性产生抑制效应,为下一步分子生物学研究奠定了基础。
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [58] |
付林俊, 刘海, 张晓晴, 张淑琴, 任大军, 2019. 不同离子对漆酶酶活的影响. 化学试剂, 41(8): 830-835
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [59] |
韩增华, 刘佳宁, 党阿丽, 张丕奇, 戴肖东, 张介驰, 2012. 黑木耳漆酶纯化及部分漆酶特性的研究. 菌物研究, 10(4): 234-239
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [60] |
胡楚霄, 雷善钰, 秦艳平, 赵奕锦, 向泉桔, 2019. 蒽对3株灵芝菌株漆酶活性及其转录表达水平的影响. 生物技术通报, 35(9): 112-117
探索多环芳烃（Polycyclic aromatic hydrocarbons,PAHs）蒽对不同灵芝菌株漆酶活性及其转录表达的影响。以3株灵芝菌株（美芝、荣保1号和川芝）为实验材料,避光静置培养5 d后,加入终浓度为1.0 mg/L的蒽,避光处理1 h、3 h、6 h、9 h和24 h,收集上清液测定漆酶活性,菌丝体样品提取RNA,分析多环芳烃蒽对漆酶活性和转录表达水平的影响。结果显示,蒽对3株菌株的漆酶活性及转录表达水平的影响均存在差异。3株灵芝菌株中,川芝的漆酶活性最高,美芝次之,荣保1号酶活最低。荣保1号漆酶活性随着处理时间的延长而上升,培养前期活性被为抑制,后期则被促进;美芝漆酶活性变化趋势类似于荣保1号,变化幅度较小;川芝漆酶活性受到强烈抑制,且随着时间的变化差异不明显。蒽处理下,美芝漆酶基因转录表达水平差异较小,大部分基因在处理9 h后出现转录表达水平峰值;荣保1号漆酶基因的表达水平类似于美芝;川芝漆酶在短期处理下,转录表达水平大幅上调,随着处理时间的延长,转录表达水平下调。3株灵芝菌株漆酶活性和漆酶基因转录表达水平对蒽的响应各不相同,其中川芝较敏感,荣保1号次之,美芝变化幅度最小,耐受力最强。
{{custom_citation.content}}
{{custom_citation.annotation}}
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| [61] |
马银鹏, 姜威, 张丕奇, 戴肖东, 周舒扬, 马庆芳, 刘佳宁, 田爽, 朱加楠, 张介驰, 王天亮, 2023. 黑木耳液体发酵条件优化研究. 食用菌, 45(1): 21-24
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [62] |
潘年港, 刘凌云, 朱姝蕊, 程国辉, 王辛, 张兆娟, 李长田, 2023. 不同遮光处理对黑木耳菌丝抗氧化及子实体生长发育的影响. 菌物学报, 42(9): 1953-1965
黑木耳Auricularia heimuer全日光栽培模式下，在出耳期持续的光照刺激使菌丝处在不适宜的生长环境。本研究采用不同遮光程度的黑菌袋，设置CK (正常菌袋，0%遮光)、T1 (50%遮光)、T2 (70%遮光)和T3 (90%遮光)共4个处理，分析黑木耳菌丝在不同遮光程度下对黑木耳子实体的农艺性状、营养品质和菌丝相关酶活性的变化。结果显示，在遮光处理下黑木耳原基形成时间相对缩短、出耳整齐、未出芽数减少，各处理第一茬产量呈显著性差异，CK处理为22.09 g，T1处理为25.20 g，T2处理为25.35 g，T3处理为25.58 g;遮光处理下不同时期菌丝的漆酶、羧甲基纤维素酶、半纤维素酶及淀粉酶活性显著高于CK处理，在采收期木质素、纤维素和半纤维的含量低，同时保持着较高的超氧化物歧化酶(superoxide dismutase, SOD)和过氧化氢酶(catalase activity, CAT)活性，丙二醛(malondialdehyde, MDA)的含量低，缓解了对机体的损伤，与CK处理呈显著性差异，其中T1处理和T2处理表现较好;遮光处理下子实体的粗蛋白含量、氨基酸含量较高，CK处理的粗脂肪含量较高，各处理存在差异。本研究重点讨论黑木耳菌丝在出耳期持续光照胁迫对子实体生长发育的影响，为黑木耳栽培提供一种更高效的栽培方式。
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [63] |
沈若茗, 陆俊佑, 张煜, 魏华, 陆玲, 2024. 大型食药用菌的降血脂功能研究进展. 菌物学报, 43(8): 4-15
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [64] |
司静, 崔宝凯, 戴玉成, 2011a. 栓孔菌属漆酶高产菌株的初步筛选及其产酶条件的优化. 微生物学通报, 38(3): 405-416
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [65] |
司静, 崔宝凯, 贺帅, 戴玉成, 2011b. 微酸多年卧孔菌产漆酶条件优化及其在染料脱色中的应用. 应用与环境生物学报, 17(5): 736-741
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [66] |
司静, 李伟, 崔宝凯, 戴玉成, 2011c. 真菌漆酶性质、分子生物学及其应用研究进展. 生物技术通报,2: 48-55
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [67] |
孙健, 2022. 黑木耳纤维素降解相关基因的挖掘及功能研究. 东北林业大学博士论文,哈尔滨. 1-126
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [68] |
唐禄鑫, 王雅娴, 彭明意, 王豪, 司静, 崔宝凯, 2023. 真菌漆酶及其生产、固定化与应用. 菌物学报, 42(9): 1821-1837
漆酶能够作用的底物非常广泛，包括木质纤维素等大分子聚合物，且在催化反应时可将该类物质还原成水和其他小分子物质，不会造成二次污染，因此，漆酶素有“绿色催化剂”之称，具有被广泛运用于工业生产中的广阔前景。对游离漆酶进行固定化处理能够使其进一步适应工业生产中的恶劣环境，提高稳定性、可循环利用率，扩大耐受pH和温度范围。真菌漆酶作为漆酶的重要来源，本文对其性质、生产，以及新型的固定化方法和各个领域的应用进行了概述。
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{{custom_citation.annotation}}
|
| [69] |
王豪, 唐禄鑫, 马鸿飞, 钱坤, 司静, 崔宝凯, 2021. 东方栓孔菌漆酶的固定化及其对不同类型染料的脱色作用. 生物技术通报, 37(11): 142-157
漆酶是一种天然的绿色催化剂,由于具有催化效率高、底物特异性广、对辅因子和特定环境条件要求少、无毒等优点,因而在制浆造纸、生物合成、改善纤维性能、食品加工、生物传感器制造、农林废弃物的生物转化和炼制,特别是环境污染物的生物降解和生物修复等领域具有巨大的应用潜力。但游离漆酶稳定性差且成本较高,固定化方法成为解决该问题的有效手段,其能够有效增强酶的热稳定性及对极端环境的耐受力、使酶易与产物分离以提高酶回收率。本研究基于前期筛选获得的一株高产漆酶白腐真菌菌株东方栓孔菌（Trametes orientalis）所分泌的漆酶（Tolacc-T）,对其纯化酶蛋白以壳聚糖作为载体、戊二醛作为交联剂进行了固定化处理,命名为Tolacc-T@Chit@GA,并优化了固定化条件为戊二醛浓度0.7%（V/V）、交联时间4 h、给酶量6.0 mL、固定化时间6 h。与Tolacc-T相比,Tolacc-T@Chit@GA的pH适应性、耐热变性能力及贮存稳定性均显著增强。循环使用时的稳定性和耐久性也十分明显,循环使用7次后,Tolacc-T@Chit@GA的相对活性仍可保持在80%以上。此外,Tolacc-T@Chit@GA还可使不同类型的染料脱色,尤其对金属络合染料萘酚绿B的脱色效果最佳,气相色谱-质谱联用（gas chromatography-mass spectroscopy,GC-MS）检测确定该染料部分代谢产物为1-萘胺、2-萘酚、1-氨基-2-萘酚、1-亚硝基-2-萘酚、1-亚硝基-2-萘酚-6-磺酸。上述结果证明,东方栓孔菌固定化漆酶Tolacc-T@Chit@GA具有较好的稳定性和可重复利用性,存在广阔的应用前景。
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [70] |
吴芳, 戴玉成, 2015. 黑木耳复合群中种类学名说明. 菌物学报, 34(4): 604-611
黑木耳在我国已经有1 000多年栽培历史,是我国最重要的栽培真菌之一,多年来我国黑木耳的学名一直使用模式产地为欧洲的Auricularia auricula-judae。最近的研究表明A. auricula-judae实际为一复合种,该复合种在全球范围内有5个种。其中A. auricula-judae仅分布于欧洲;美洲有2个种,生长在针叶树上的美洲木耳A. americana和生长在阔叶树上的尚未命名的木耳。中国该类群有3个种：自然分布和栽培最广泛的黑木耳的学名应为A. heimuer,此外,短毛木耳A. villosula在我国东北也广泛分布,并有少量栽培,生长针叶树上的木耳为美洲木耳A. americana,主要分布于中国东北和华北。
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [71] |
吴怡, 马鸿飞, 曹永佳, 司静, 崔宝凯, 2019. 真菌漆酶的性质、生产、纯化及固定化研究进展. 生物技术通报, 35(9): 1-10
真菌漆酶是一种性质优良的多酚氧化酶,由于在分子氧的协助下可将酚类、芳胺类化合物等多种底物氧化,最终得到水及其终产物,符合当代环保工业要求,因而在纸浆漂白、环境治理、生物检测、有机合成等领域有着巨大的应用潜力。就漆酶的生物学性质、生产、纯化、固定化等研究进展和现状进行了介绍和总结,同时对其今后的发展方向进行了展望。
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{{custom_citation.annotation}}
|
| [72] |
吴怡, 马鸿飞, 曹永佳, 司静, 崔宝凯, 2020. 白腐真菌落叶松锈迷孔菌产漆酶液体培养基的优化及其对染料的脱色作用. 生物技术通报, 36(1): 45-59
以白腐真菌落叶松锈迷孔菌（Porodaedalea laricis）胞外漆酶为响应值,通过将Plackett-Burman设计、最陡爬坡设计和Box-Behnken设计相结合,获得了P. laricis产胞外漆酶的最适培养基为：去皮马铃薯365.61 g/L、蛋白胨5.0 g/L、葡萄糖20.0 g/L、KH<sub>2</sub>PO<sub>4</sub> 1.0 g/L、MgSO<sub>4</sub>·7H<sub>2</sub>O 0.5 g/L、MnSO<sub>4</sub>·H<sub>2</sub>O 0.15 g/L、CaCl<sub>2</sub>·2H<sub>2</sub>O 0.03 g/L、酒石酸铵6.68 g/L、琥珀酸钠1.5 g/L、吐温80 0.48 mL/L、玉米芯46.43 g/L、维生素B1 0.01 g/L。在该条件下,P. laricis漆酶活性为3.29 U/mL,相比于优化前提高了2.81倍,与理论值3.32 U/mL相近,说明该模型准确可靠。此外,将漆酶应用于降解多种合成染料包括活性亮蓝X-BR、雷马素亮蓝R、酸性黑172、刚果红、亚甲基蓝、中性红、靛蓝、萘酚绿B和结晶紫,反应168 h后脱色率分别可达到95.64%、97.21%、36.11%、91.63%、61.42%、74.65%、48.60%、25.13%和68.80%。
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [73] |
肖楚, 2012. 黑木耳漆酶高产菌株筛选及发酵条件、酶学性质的研究. 东北农业大学硕士论文,哈尔滨. 1-79
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [74] |
杨迪, 2023. 玉米芯栽培黑木耳的配方筛选及胞外酶活性研究. 吉林农业大学硕士论文,长春. 1-56
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [75] |
余卓, 2021. 蓝铜胜肽诱导变色栓菌液态发酵产漆酶及其机制的研究. 江苏大学硕士论文,镇江. 1-91
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [76] |
张津京, 陈明杰, 冯志勇, 黄建春, 宋晓霞, 汪虹, 陈辉, 2016. 芳香族化合物对斑玉蕈菌丝生物量、漆酶活性及其转录水平的影响. 菌物学报, 35(9): 1130-1138
芳香族化合物适当时间适当浓度添加到培养基中,可提高真菌漆酶活性,有助于增强其对木质纤维素的利用效率。为了增强斑玉蕈漆酶活性,本文研究了8种芳香族化合物对其酶活的影响及其与菌丝生物量的相关性。研究发现在无诱导物条件下,斑玉蕈漆酶活性和菌丝生物量相关系数r为0.9956,说明它们呈正相关,但是整个培养过程漆酶活性相对较低;供试的芳香族化合物对漆酶活性都有不同程度的诱导作用,其中添加0.1mmol/L的愈创木酚对斑玉蕈漆酶活性诱导作用最大,达到3倍以上,同时提高了斑玉蕈菌丝生长速度和菌丝生物量;而随着添加时间的延长,部分化合物对漆酶活性和菌丝生物量都产生不同程度的抑制作用,这可能因为化合物对菌丝毒性的延长导致菌丝生长变慢或死亡;进一步研究发现,斑玉蕈3个漆酶同工酶基因lcc2、lcc3和lcc4在诱导剂愈创木酚的影响下转录水平都不同程度地上调。研究结果表明诱导漆酶活性可以提高斑玉蕈菌丝生长速度和生物量,暗示可能通过提高漆酶活性的方法,提高斑玉蕈的培养基利用效率。
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [77] |
赵丽婷, 2017. 共培养胶红酵母对阿魏蘑漆酶基因表达及其相关蛋白的影响. 江南大学硕士论文,无锡. 1-51
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [78] |
卓睿, 2015. 白腐真菌漆酶及同工酶基因家族的克隆表达调控研究. 华中科技大学博士论文,武汉. 1-197
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Table 1 Content of main active components in CME
Fig. 1 Determination of monosaccharide composition by HPLC. A: Mixed reference substance; B: Tested samples. 1: Man; 2: Rha; 3: Glc-UA; 4: Gal-UA; 5: Lactose (internal standard); 6: Glc; 7: Gal: 8: Xyl; 9: Ara; 10: Fuc.Table 2 Monosaccharide composition of Cordyceps militaris polysaccharideFig. 2 Determination of nucleosides by HPLC. A: Mixed reference substance; B: Test sample. 1: Cytidine; 2: Uridine; 3: Guanosine; 4: Thymidine; 5: Adenosine; 6: Cordycepin; 7: N6-(2-hydroxyethtl)- adenosine.Fig. 3 Scavenging effects of CME on radicals. A: DPPH radical scavenging activities; B: ABTS radical scavenging activities; C: ·OH radical scavenging activities.Fig. 4 Effects of different concentrations of CME on the viabilities of HaCaT cells.Fig. 5 Effects of different concentrations of H2O2 on the viabilities of HaCaT cells.Fig. 6 Effects of different concentrations of CME on the viabilities of HaCaT cells induced by H2O2. Compared with control group, ##P<0.01; Compared with model group, *P<0.05, **P<0.01. The same below.Fig. 7 Effects of different concentrations of CME on ROS levels in HaCaT cells.Fig. 8 Effects of different concentrations of CME on MDA contents (A) and the activities of SOD (B), GSH-Px (C), CAT (D) in HaCaT cells.
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