海洋微生物来源的α-葡萄糖苷酶抑制剂的筛选及性质研究

建立了α-葡萄糖苷酶抑制剂的体外筛选模型。10株海洋微生物经摇床培养得到发酵液,经大孔树脂吸附、甲醇洗脱得到粗提物;以阿卡波糖为阳性对照,用比色法（PNPG法）对粗提物进行α-葡萄糖苷酶抑制活性测定,并对产生活性代谢产物的菌株做进一步研究。结果发现：有6株菌株的代谢产物均具有不同程度的α-葡萄糖苷酶抑制作用,其中菌株N-1的代谢产物对α-葡萄糖苷酶的抑制作用最强,每毫升N-1样品液中含有的活性代谢产物对α-葡萄糖苷酶的抑制作用相当于5mg的阿卡波糖对α-葡萄糖苷酶的抑制作用;该活性产物具有较强的酸碱耐受性及温度耐受性,在pH值1～14、50～100℃时稳定性较好;初步分析活性代谢产物的成分并非蛋白质、氨基酸、糖类及皂苷类物质。     

菌株H008发酵条件的优化及其发酵液杀虫活性物质的研究

对菌株H008(Streptomyces laidensis)的发酵培养条件进行了优化,并对其发酵产物的生物学活性进行了初步研究.采用正交试验法,以菌株H008发酵液的杀虫活性为指标进行了培养条件的优化.通过正交试验确定的发酵生产工艺参数为pH值8.0、发酵温度28℃、溶解氧60%、发酵周期96 h.发酵液中该菌株的代谢产物具有杀虫活性高、热稳定性高、酸碱适应范围广、极性强等特性.     

一株盐湖嗜盐菌的培养条件研究

嗜盐放线菌研究是极端环境微生物研究的重要组成部分,选择性采用高盐极端环境放线细菌新颖菌株进行菌株多种方式培养及次生代谢产物研究,有望找到新的有用化学结构.文章对一株分离自盐湖嗜盐菌的培养条件进行了研究,通过其平板及斜面培养情况、发酵培养情况、薄层层析技术显色等最终确定了其较为合适的培养基成分.     

青岛海域鲍鱼内生真菌代谢产物活性筛选探究

以开放实验的形式探究海洋内生真菌的分离纯化实验操作,锻炼学生微生物实验技术中的无菌操作、菌种的分离纯化过程,进而通过发酵条件的优化培养对海洋真菌的次级代谢产物进行初步的研究,使学生进一步了解环境微生物中真菌发酵条件的调控和次级代谢产物的应用。通过微生物开放实验分离纯化出4株来源于鲍鱼的海洋内生真菌A1、A2、A3和A4,选择菌株A1通过优化培养发现其RICE培养基的次级代谢产物量较大,且有较好的抑菌活性。开放实验既提高了学生科技创新意识,又使学生接受了初步的科研训练。     

培养基组成成分对加利利链霉菌AF1发酵过程影响的初步探索

探索影响加利利链霉菌AF1发酵过程的培养基组成成分。碳、氮源试验结果表明菌株AF1能够较好的利用可溶性淀粉和胰蛋白胨。在基础培养基和培养条件不变的前提下,改变可溶性淀粉质量浓度,在质量浓度为3.5%时,菌株AF1的代谢产物对金黄色葡萄球菌的抑菌活性最高,达到15.5 mm。在基础培养基和培养条件不变的前提下,改变胰蛋白的质量浓度,当浓度达到0.25%时,抑菌活性最高,达到13.4 mm。供试的8种氨基酸试验结果表明Arg、Gln和Lys具有明显促进菌体生长的作用,Asn、Phe的促进作用不明显,而其他的氨基酸则表现出对菌体生长无显著效果;在产物合成方面,Arg、Asn、Phe、Tyr、Gln、Lys对产物合成具有较为明显的促进作用,但Met和Cys却表现出对产物合成有抑制作用。不同浓度的各种氨基酸试验结果表明,当基础培养基中Tyr浓度为0.13 mmol/L或者Lys 0.25 mmol/L时菌株生长和活性物质的代谢效果最为明显,100 mL的发酵液中菌体干重和发酵液抑菌活性分别达到0.19 g、14 mm和0.22 g、15.9 mm。     

真菌诱导子对悬浮培养南方红豆杉细胞次生代谢的影响

在悬浮培养的南方红豆杉细胞体系中 ,细胞进入指数生长期末期时 ,加入真菌诱导子 (即尖孢镰刀菌胞壁成分的粗提出物 ,其主要成分为糖类和多肽 ) ,对几个主要的细胞次生代谢指标进行了监测 .实验结果表明 ,诱导子引起了南方红豆杉细胞次生代谢结构的变化 ,胞内酶蛋白的含量呈先上升后下降的趋势 ,而胞外酶蛋白的含量显著上升 ;苯丙烷类代谢途径中重要的酶PAL的活性显著升高 ;次生代谢产物中 ,酚类物质的含量增加 ,且于 4天后急速上升 ,同时多酚氧化酶的活力也加强 ,萜类产物中紫杉烷类的生物合成均得到了加强 ,其中紫杉醇的含量得到了大幅度的提高 ,达到对照组的 5倍左右 .     

铜绿假单胞菌次生代谢产物的研究进展

铜绿假单胞菌可产生多种具有活性的次生代谢产物。综述了铜绿假单胞菌次生代谢产物的种类、提取方法、合成方法、作用机制及用途,指出了研究中存在的问题,展望了未来的发展方向,拟为铜绿假单胞菌次生代谢产物的科学研究提供参考。     

腐植酸应用在酿酒工业的初步研究

酿酒工业的发酵产品是微生物的代谢产物,如何给微生物创造适宜的生长、生活条件,除温度水份外,关键决定于培养基成分和工艺控制,我们对腐植酸增效剂(以下简称增效剂)的研究和应用,目的就是调整培养基成分,改善微生物的生活条件,促使微生物在生化过程中发挥更大的作用,从而获得更多的代谢产物和经济效果。去年以来,     

北极海泥来源抗菌活性真菌的筛选及菌株H5的初步研究

从北极海泥中筛选抗茵活性海洋真菌,鉴定了活性菌株并对活性最高的菌株H5的代谢产物进行了初步研究。通过形态学观察、基因间隔区（ITS）序列分析对活性最高的茵株H5进行系统发育分析和菌种鉴定;通过纸片法对茵株H5的发酵液进行体外抗茵活性研究;将调至不同pH值的H5发酵液用不同极性的有机溶剂萃取,研究该活性物质的酸碱性和极性;将调至不同pH值的发酵液按不同时间温度梯度进行加热处理,研究该活性物质的稳定性;同时用大孔树脂对活性物质进行提取分离。结果表明,从北极海泥中共分离得到17株真菌,其中真菌H5、H6的代谢产物具有较好的抗茵活性;茵种鉴定表明菌株H5归属于烟曲霉属（Aspergillusfumigatus）,该菌株发酵液对苹兰氏阳性菌、革兰氏阴性茵、真茵及耐药茵均有不同程度的抑制活性,活性产物为极性较大、热稳定性较好、耐酸不耐碱的物质;薄层层析显示活性成分在254rlm紫外光下无吸收。     

通过统计优化培养条件增强链霉菌S-159的抗真菌活性

目的：采用统计实验方法优化培养条件,以增加链霉菌S-159抗真菌活性成分的产量。方法：首先采用PlackettBurman设计以评估11个培养条件（包括2个虚拟变量）的显著性,发现大豆粉添加量、葡萄糖添加量、发酵时间和初始p H值为影响抗真菌活性的最显著因素。接下来通过最速上升路径分析确定进一步优化的中心点,并使用响应面法（RSM）中的中心复合设计（CCD）进一步优化获得的4个显著因素。结果：实验结果符合二阶多项式模型,其确定系数（R2）为0.986,调整后的确定系数（Adj R2）为0.974。求解方程得到优化组合条件为大豆粉添加量23.53 g/L、葡萄糖添加量17.04 g/L、发酵时间60.20 h和初始pH值7.61,抗真菌活性实测值为410.5 U/mL,与该模型预测的413.6 U/mL基本一致,优化后链霉菌S-159代谢产物的抗真菌活性提高了1.7倍。结论：统计实验方法可用于链霉菌S-159的发酵培养条件优化,研究结果为链霉菌S-159的大范围应用提供了数据基础。     

Antifungal leaf-surface metabolites correlate with fungal abundance in sagebrush populations

A central component in understanding plant–enemy interactions is to determine whether plant enemies, such as herbivores and pathogens, mediate the evolution of plant secondary metabolites. Using 26 populations of a broadly distributed plant species, sagebrush (Artemisia tridentata), we examined whether sagebrush populations in habitats with a greater prevalence of fungi contained antifungal secondary metabolites on leaf surfaces that were more active and diverse than sagebrush populations in habitats less favorable to fungi. Because moisture and temperature play a key role in the epidemiology of most plant–pathogen interactions, we also examined the relationship between the antifungal activity of secondary metabolites and the climate of a site. We evaluated the antifungal activity of sagebrush secondary metabolites against two fungi, a wild Penicillium sp. and a laboratory yeast, Saccharomyces cerevisiae, using a filter-paper disk assay and bioautography. Comparing the 26 sagebrush populations, we found that fungal abundance was a good predictor of both the activity (r ² = 0.36 for Saccharomyces, r ² = 0.37 for Penicillium) and number (r ² = 0.34 for Saccharomyces) of antifungal secondary metabolites. This suggests that selection imposed by fungal pathogens has led to more effective antifungal secondary metabolites. We found that the antifungal activity of sagebrush secondary metabolites was negatively related to average vapor pressure deficit of the habitat (r ² = 0.60 for Saccharomyces, r ² = 0.61 for Penicillium). Differences in antifungal activity among populations were not due to the amount of secondary metabolites, but rather to qualitative differences in the composition of antifungal compounds. Although all populations in habitats with high fungal prevalence had secondary metabolites with high antifungal activity, different suites of compounds were responsible for this activity, suggesting independent outcomes of selection on plants by fungal pathogens. The location of antifungal secondary metabolites on the leaf surface is consistent with their putative defense role, and we found no evidence supporting other functions, such as protection from ultraviolet light or oxidation. That the antifungal activity of sagebrush secondary metabolites was similar for two different fungi provides support for broad antifungal defenses. The incidence and severity of fungal disease in the field (caused by Puccinia tanaceti) were similar in moist and dry habitats, possibly reflecting an equilibrium between plant defense and fungal attack, as sites with greater fungal abundance compensated with more effective secondary metabolites. The geographic correlation between fungal abundance and antifungal secondary metabolites of sagebrush, coupled with our other data showing heritable variation in these metabolites, suggests that pathogenic fungi have selected for antifungal secondary metabolites in sagebrush.     

Antifungal Diketopiperazines from Symbiotic Fungus of Fungus-Growing Ant Cyphomyrmex minutus

The attine fungus Tyridiomyces formicarum, the symbiont of the fungus-growing ant Cyphomyrmex minutus, produces several antifungal diketopiperazines. This represents the first identification of antifungal compounds from an attine symbiont and contradicts previous suggestions that attine fungi do not produce metabolites with antifungal activity. T. formicarum probably produces antifungal compounds in defense (1) against other fungi that invade the gardens and escape the weeding activity of the ants, or (2) against ant-pathogenic fungi that could harm the host ants. Fungi cultivated by fungus-growing ants may represent a rich source of additional bioactive metabolites.     

海洋微生物源农用抗生活性物质的筛选

对134株海洋微生物(其中49株海洋放线菌、54株海洋细菌、31株海洋真菌)进行农用抗生活性物质筛选,分别测定其发酵上清液和菌丝体浸提液的活体抗植物病源真菌、杀虫、除草活性,发现共有77株菌株具有农用抗生活性,其中7株海洋放线菌的代谢产物的活性较高:3株对植物病原真菌具有较好抑制活性的海洋放线菌代号为Y12–26、P10–16、JWH–09;两株具有较高杀蚜虫活性的海洋放线菌是P10–23、P12–21;两株除草活性较好的海洋放线菌是YY01–17和HH1901–45。     

Imaging Mass Spectrometry of a Coral Microbe Interaction with Fungi

Fungal infections are increasing worldwide, including in the aquatic environment. Microbiota that coexist with marine life can provide protection against fungal infections by secretion of metabolites with antifungal properties. Our laboratory has developed mass spectrometric methodologies with the goal of improving our functional understanding of microbial metabolites and guiding the discovery process of anti-infective agents from natural sources. GA40, a Bacillus amyloliquefaciens strain isolated from an octocoral in Panama, displayed antifungal activity against various terrestrial and marine fungal strains. Using matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS), the molecular species produced by this microbe were visualized in a side-by-side interaction with two representative fungal strains, Aspergillus fumigatus and Aspergillus niger. The visualization was performed directly on the agar without the need for extraction. By evaluating the spatial distributions, relative intensities and m/z values of GA40 secreted metabolites in the fungal interactions and singly grown control colonies, we obtained insight into the antifungal activity of secreted metabolites. Annotation of GA40 metabolites observed in MALDI-IMS was facilitated by MS/MS networking analysis, a mass spectrometric technique that clusters metabolites with similar MS/MS fragmentation patterns. This analysis established that the predominant GA40 metabolites belong to the iturin family. In a fungal inhibition assay of A. fumigatus, the GA40 iturin metabolites were found to be responsible for the antifungal properties of this Bacillus strain.     

海洋放线菌Y-0117农用活性代谢产物的研究

在农用抗生素筛选中发现代号为Y-0117的海洋放线茵菌株所产生的代谢产物对植物病原真菌具有良好的抑制作用。对Y-0117发酵液进行萃取、硅胶柱层析、制备薄层色谱、制备HPLC,分离纯化得到两个活性组分0117A和0117B。通过紫外光谱、质谱、核磁共振等手段,确定0117A的分子式为C35H58O8,分子量为604。结构与已知化合物Bafilomycin D相同;0117 B的分子式为C42H60O12．分子量为756,结构与近期发现具有抗癌活性的已知化合物Hygrobafilomycin相同。研究发现0117B对多种植物病原真菌具有良好的抑制活性,对稻瘟病菌的最小抑制浓度为15．63μg·mL^-1。     

丹皮酚合成过程中抑菌活性的变化研究

研究了在丹皮酚的合成过程中,原料间苯二酚、中间产物2,4-二羟基苯乙酮及主产物丹皮酚对11种植物病原真菌的抑菌活性,并通过抑菌活性的变化来推测影响抑菌活性的主要官能团.结果表明:0.5g·L-1的2,4-二羟基苯乙酮对11种真菌的抑菌率大于或等于同浓度丹皮酚的抑菌率,两者均远远大于同浓度间苯二酚的抑菌率.由此可推测,酚类分子中乙酰基的引入对抑菌活性起到了非常重要的作用,而酚羟基的甲基化的作用却并不显著.该结论对寻求具有更高抑菌活性的酚类衍生物有着非常重要的意义.     

A study on melt spinning of polyethylene terephthalate filament yarns containing various concentrations of synthesized CuO nanoparticles

In this research the CuO nanoparticles were synthesized with four different methods then used to produce polyethylene terephthalate (PET) fibers with antibacterial, antifungal, and photocatalytic properties during the melt spinning process. Nanoparticles were synthesized in different ways using copper acetate salt (Cu(CH₃COO)₂) and sodium hydroxide (NaOH). The synthesis of copper oxide (CuO) nanoparticles was confirmed by field-emission scanning electron microscopy (FE-SEM images), EDX patterns, TGA, FTIR, X-ray diffraction pattern XRD, and UV-vis absorption spectrum. The results showed that synthesized nanoparticles exhibited strong antibacterial activity against gram-positive and gram-negative bacteria. Finally, after examining the fibers with different percentages of nanoparticles), fibers containing 1% of nanoparticles were produced using a 20% nanocopper oxide masterbatch as the optimal sample. The presence of nanoparticles in the fiber structure was confirmed by FE-SEM images, EDX, XRD, and FTIR analysis. Furthermore, the tensile properties, thermal behavior, and photocatalytic properties of the fibers were investigated. Evaluation of the antibacterial and antifungal activity of the produced fibers against gram-positive bacteria Staphylococcus aureus and Candida albicans fungus showed about 90% death of bacteria and fungi.     

Secondary Metabolites of the Rice Blast Fungus Pyricularia oryzae: Biosynthesis and Biological Function

Plant pathogenic fungi produce a wide variety of secondary metabolites with unique and complex structures. However, most fungal secondary metabolism genes are poorly expressed under laboratory conditions. Moreover, the relationship between pathogenicity and secondary metabolites remains unclear. To activate silent gene clusters in fungi, successful approaches such as epigenetic control, promoter exchange, and heterologous expression have been reported. Pyricularia oryzae, a well-characterized plant pathogenic fungus, is the causal pathogen of rice blast disease. P. oryzae is also rich in secondary metabolism genes. However, biosynthetic genes for only four groups of secondary metabolites have been well characterized in this fungus. Biosynthetic genes for two of the four groups of secondary metabolites have been identified by activating secondary metabolism. This review focuses on the biosynthesis and roles of the four groups of secondary metabolites produced by P. oryzae. These secondary metabolites include melanin, a polyketide compound required for rice infection; pyriculols, phytotoxic polyketide compounds; nectriapyrones, antibacterial polyketide compounds produced mainly by symbiotic fungi including endophytes and plant pathogens; and tenuazonic acid, a well-known mycotoxin produced by various plant pathogenic fungi and biosynthesized by a unique NRPS-PKS enzyme.     

Rhizopus and Fusarium are Selected as Dominant Fungal Genera in Rhizospheres of Brassicaceae

We isolated several strains of Rhizopus and Fusarium spp. as dominant fungi in the rhizospheres of Brassicaceae plants. The Fusarium isolates showed a higher tolerance of the antifungal constituents of "mustard oil," which originates from the glucosinolates that are characteristic secondary metabolites of the Brassicaceae, than other Fusarium isolates from non-Brassicaceae plants. In contrast, the Rhizopus isolates showed a high tolerance regardless of their source. Myrosinase activity was found in Bn-R-1-1 (Rhizopus sp.) isolated from the rhizoplane of Brassica napus and Ls-F-in-4-1 (Fusarium sp.) isolated from a surface-disinfected root of Lepidium sativum (Brassicaceae). Ls-F-in-4-1 was the Fusarium most tolerant of the Brassicaceae antifungal constituents. These results suggest that fungi in the rhizospheres of Brassicaceae plants may be selected because of secondary metabolites exuded from the roots of host plants.     

新型抗真菌抗生素发酵工艺及动力学研究

从浙江省东海土壤中筛选到1株具有抑制白色念珠菌ATCC 64548生长的解淀粉芽孢杆菌091,发现其能产生一种相对分子质量为410的大环内酯类抗真菌物质。在250 mL摇瓶中进行了抗生素发酵的单因素优化实验,实验结果:28℃,pH值6.5,摇床转速200 r/min,接种量(体积分数)8%,装液量40 mL,发酵时间36 h。优化后发酵液抗真菌效价比初始培养条件下提高了2.49倍,达7 026.79 U/mL。进一步采用Logistic,Luedeking-Piret和Lu-edeking-Piret-like方程建立了抗生素产生的动力学模型,并根据5 L发酵罐中的实验数据拟合得到了模型参数。结果表明:该数学模型能反映解淀粉芽胞杆菌091的细胞生长、底物消耗和抗生素合成的动力学机制,可为该抗真菌抗生素的中试生产提供理论指导。