江西三种危害食用菌的绿色霉菌生物学特性研究

本文报道了江西三种危害食用菌的绿色霉菌的生物学特性。它们是:绿色木霉(Trichoderma Viride pers exFn)。产黄青霉(Penicillium Chrysoenum Thom)和构巢曲霉(Aspergillus nidulcms(Eidzm)Winter)。     

花生抗黄曲霉相关基因PnLOX2的原核表达及RNAi载体的构建与转化

本研究对利用基因芯片技术从抗、感黄曲霉花生品种中分离出的差异表达基因PnLOX2进行原核表达,得到分子量为121.5 kD的融合表达产物;构建了PnLOX2基因的3'端反向重复结构并转化花生,得到了转基因花生苗,为反向鉴定PnLOX2基因在花生抗黄曲霉侵染过程中的功能奠定基础。     

花生黄曲霉生长预测模型的研究

通过对分离自花生的黄曲霉LYYSS03-3在不同温度(15-35℃)、不同相对湿度(水活度分别为0.5,0.7,0.9)下的菌落生长特性进行检测,采用Boltzmann和Logistic 模型对黄曲霉的生长曲线进行拟合,结果表明,在适宜温湿度阶段,花生黄曲霉的生长曲线更适合用Boltzmann模型拟合.     

黑曲霉菌渣资源化再利用途径分析

黑曲霉菌渣是黑曲霉发酵法生产葡萄糖酸钠后产生的废弃物，其资源化利用已成为企业扩大生产亟需解决的问题。对该菌渣有效成分检测得知，该菌渣有害金属元素均小于城镇垃圾农用控制标准、有机-无机复混肥料控制标准和鱼粉、猪、家禽添加剂预混合饲料卫生指标，有机质含量符合有机肥料中的有机质含量指标，总糖、粗纤维、粗蛋白、脂肪等营养成分含量较多，同时也含有丰富的N、P、K、Ca、Mg、Fe等元素，可从有机肥料和禽畜饲料方向进行资源化利用，有利于减轻环境污染及实现变废为宝，具有积极的社会、经济和生态效益。     

凉茶渣的黑曲霉发酵工艺和抗氧化活性研究

为促进凉茶渣在畜牧业中的资源化利用,本研究以黑曲霉为菌种固态发酵凉茶渣,首先在单因素试验条件下考察了时间、温度、含水量、浸泡液pH、氮源和碳源对凉茶渣降解率和产物pH的影响;再根据单因素试验结果和规模化生产实际需要,以4%硫酸铵为氮源,2%葡萄糖为碳源,以降解率为考察指标,通过正交试验优化发酵工艺;并通过检测超氧自由基清除率、羟基自由基清除率和1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除率,评价凉茶渣在最优工艺条件下发酵前后抗氧化活性的变化。结果发现,含水量为60%,浸泡液pH为9.0,31℃发酵168 h是凉茶渣的最佳发酵工艺参数。最佳工艺条件下凉茶渣的降解率为25.23%,发酵产物pH为4.53。当凉茶渣发酵前水提液浓度为24 mg/mL时,超氧自由基清除率为43.56%,羟基自由基清除率为47.06%,DPPH自由基清除率为90.71%;当最优条件下发酵产物水提液浓度24 mg/mL时,超氧自由基清除率为30.77%,羟基自由基清除率为95.63%,DPPH自由基清除率为87.36%。本试验结果表明,黑曲霉是适宜的凉茶渣发酵菌种,且凉茶渣经过黑曲霉发酵后具有良好的抗氧化活性。     

Aurasperone A Inhibits SARS CoV-2 In Vitro: An Integrated In Vitro and In Silico Study

Several natural products recovered from a marine-derived Aspergillus niger were tested for their inhibitory activity against SARS CoV-2 in vitro. Aurasperone A (3) was found to inhibit SARS CoV-2 efficiently (IC₅₀ = 12.25 µM) with comparable activity with the positive control remdesivir (IC₅₀ = 10.11 µM). Aurasperone A exerted minimal cytotoxicity on Vero E6 cells (CC₅₀ = 32.36 mM, SI = 2641.5) and it was found to be much safer than remdesivir (CC₅₀ = 415.22 µM, SI = 41.07). To putatively highlight its molecular target, aurasperone A was subjected to molecular docking against several key-viral protein targets followed by a series of molecular dynamics-based in silico experiments that suggested M^pro to be its primary viral protein target. More potent anti-SARS CoV-2 M^pro inhibitors can be developed according to our findings presented in the present investigation.     

Neoechinulin A as a Promising SARS-CoV-2 Mpro Inhibitor: In Vitro and In Silico Study Showing the Ability of Simulations in Discerning Active from Inactive Enzyme Inhibitors

The COVID-19 pandemic and its continuing emerging variants emphasize the need to discover appropriate treatment, where vaccines alone have failed to show complete protection against the new variants of the virus. Therefore, treatment of the infected cases is critical. This paper discusses the bio-guided isolation of three indole diketopiperazine alkaloids, neoechinulin A (1), echinulin (2), and eurocristatine (3), from the Red Sea-derived Aspergillus fumigatus MR2012. Neoechinulin A (1) exhibited a potent inhibitory effect against SARS-CoV-2 M^pro with IC₅₀ value of 0.47 μM, which is comparable to the reference standard GC376. Despite the structural similarity between the three compounds, only 1 showed a promising effect. The mechanism of inhibition is discussed in light of a series of extensive molecular docking, classical and steered molecular dynamics simulation experiments. This paper sheds light on indole diketopiperazine alkaloids as a potential structural motif against SARS-CoV-2 M^pro. Additionally, it highlights the potential of different molecular docking and molecular dynamics simulation approaches in the discrimination between active and inactive structurally related M^pro inhibitors.     

Effects of wood char and activated carbon on the hydrolysis of cellobiose by β-glucosidase from Aspergillus niger

Biochar amendments to soils have been suggested as a strategy to sequester carbon and therefore mitigate global climate change. The enrichment of soils with charred materials also increases their fertility. This fertilising effect of biochar may be caused by various mechanisms; an acceleration of nutrient cycling has been suggested as one such mechanism. The rate-limiting step in nutrient cycling is thought to be the extracellular enzymatic attack on biological macromolecules. In this study, therefore, the effects of chestnut wood char (specific surface area 2.0 m² g⁻¹) and of activated carbon (specific surface area approximately 900 m² g⁻¹) on an extracellular enzymatic reaction involved in the degradation of cellulose (i.e., hydrolysis of cellobiose by β-glucosidase from Aspergillus niger) were investigated. Cellobiose was not adsorbed by chestnut wood char, whereas activated carbon absorbed more than 97% of it. Both charred materials adsorbed more than 99% of β-glucosidase. For chestnut wood char, adsorption of the enzyme caused a decrease of approximately 30% in the reaction rate, whereas for activated carbon, the nearly complete absorption of both substrate and enzyme entirely inhibited the reaction. These results show that β-glucosidase from A. niger retains most of its activity when adsorbed to chestnut wood char and that the reaction it catalyses in nature is only slightly affected by this charred material. On the other hand, a material characterised by a high specific surface area and high porosity, such as activated carbon, can make even a highly soluble substrate unavailable for soil enzymes and therefore completely inhibit the reaction. Thus, charred materials may affect nutrient cycling mainly by regulating the availability of substrates: the degradation of highly soluble substrates may be accelerated by materials with low specific surface area, which maintain an active and protected enzyme pool, whereas materials with high specific surface and high porosity may slow down the degradation by making substrates unavailable.     

一株曲霉的一种中温糖化酶的纯化及其部分酶学特性

本研究从广西花坪自然保护区采集的土壤中筛选获得了一株产糖化酶丝状真菌菌株57-45,通过形态学观察和真菌内转录间隔区（internal transcribed spacer,ITS）序列比对分析,将其初步鉴定为曲霉属（Aspergillus sp.）的一个种。纯化真菌57-45所产的一种胞外糖化酶经过硫酸铵分级沉淀、疏水层析和阴离子交换层析三步蛋白质纯化步骤,得到在SDS-PAGE上约60kD的单一蛋白质条带,薄层层析分析表明该纯化的蛋白质水解可溶性淀粉的产物只有葡萄糖,证明该纯化的蛋白质为糖化酶。纯化的糖化酶Km值为1.9mg/mL,Vmax为4148μmol/（min·mg）,最适作用pH5.5,最适作用温度50℃,在同步糖化发酵中有应用的潜力。金属离子Fe3＋、Zn2＋、Cu2＋对酶活有较强的抑制作用,EDTA对酶活有较强的促进作用。本文结果将为进一步研究曲霉糖化酶的酶学特性提供新的材料。     

高效降解有机磷农药真菌的分离鉴定及其特性研究

从有机磷农药严重污染的土壤中分离筛选出有机磷农药高效广谱降解真菌,并对其进行鉴定及降解特性研究。通过有机磷农药驯化培养、形态学和生理生化特性及其18S rDNA序列分析等试验对其进行初步鉴定,研究外界因素初始pH、培养温度、氧化乐果初始浓度等对降解菌的生长量和降解能力的影响。通过驯化分离筛选等过程从污染土壤中筛选到2株可以在高浓度氧化乐果环境下生长的真菌J4和J6,它们经生理生化和18 S rDNA序列测定初步鉴定均为黑曲霉（Aspergillus niger）。研究发现,J4和J6菌株在氧化乐果初始浓度1000 mg/L,pH值分别为6.5和6.0,温度分别为30℃和28℃,摇床转速160 r/min条件下,培养5天后降解菌可达到最佳生长量和降解能力,降解率可分别达77.83%和70.81%,耐受氧化乐果的最大浓度均为5000 mg/L,其生长量和降解能力具有正比例相关性变化趋势;并进一步研究发现,曲霉J4和J6还具有广谱降解有机磷农药的特性。