Microorganisms surviving for 5300 years

Abstract Recently, the well-preserved corpse of a prehistoric man with an age of approximately 5300 years bp was discovered in the Central European Alps. Analysis of materials associated with the individual has revealed the presence of microorganisms which are believed to have survived since the time of death. So far, two fungi have been isolated and identified as species of the genera Chaetomium and Absidia , respectively. In addition, we have obtained one bacterial isolate which we have identified as a Streptomyces species. Our findings demonstrate that microorganisms can remain viable under appropriate circumstances for thousands of years. The isolates may enable us to study evolutionary trends within microorganisms.     

关于保留球毛壳与橄榄色毛壳作为不同种的见解

球毛壳(Chaetomium globosum Kze.)系 Kunze 于1817年报道见于丹麦石竹(Dianthus carthusianorum L.)茎上的毛壳菌属(Chaetomium)的第一个种(模式种)。Cooke and Ellis 在1878年描述了见于飞蓬属(Erigeron L.)腐茎上的橄榄色毛壳(C.oliaceum C.et E.)。在 Chivers(1915)、Skolko and Groves(1953)、Udagawa(1960)、Ames(1963)和 Seth(1972)关于毛壳菌属的专著都曾指出很难划分这两个种的界限。Skolk and Groves(1953)区分此两个种时以橄榄色毛壳具有较大的子囊壳、较宽的顶附属丝和较大的子囊孢子,而 Chivers(1915)则认为橄榄色毛壳是球毛壳的异名,Udagawa(1960)区分此两个种仅根据子囊孢子的长度和宽度,他认为橄榄色毛壳的子囊孢子大于球毛壳的子囊孢子。Seth(1972)在他的专著中虽保留此两个种作为独立种,但他指出限于他镜检过的标本材料及根据 Chivers 专著中的球毛壳的特征辑要概括了橄榄色毛壳的特征,对这两个种的区分界限确实是很难划分的。最近我们在北京采集和分离了来源于不同的植物和动物材料上的毛壳菌种类,以期进行北京地区毛壳菌种类调查研究。我们分离获得许多球毛壳——橄榄色毛壳类的毛壳菌菌株。参考了不同作者对这两个种的子囊壳、顶附属丝、侧附属丝、子囊及子囊孢子的特征记载,对北京的这一类型菌株进行了细致研究,认为球毛壳与橄榄色毛壳确有形态学特征区别,表现在橄榄色毛壳的子囊壳、顶附属丝和子囊孢子较球毛壳的更为粗壮,兼之球毛壳的顶附属丝较橄榄色毛壳的为窄且有分隔和微粗糙,球毛壳的子囊孢子呈浅橄榄褐色至暗橄榄褐色,含两个折光性油滴而橄榄色毛壳的子囊孢子呈暗橄榄褐色,量度亦较大,凭依经验即可鉴别此两个不同种。     

Germinability and fungal occurrence in seeds of Abies alba Mill. populations in southern Italy

Abstract

Germination tests were carried out on seeds from three Italian populations of Abies alba. Seed germinability was up to 38%. HgCl₂, NaClO and plant preservative mixture were used as seed‐sterilising agents. Despite chemical treatments, Papulaspora rubida and Chaetomium globosum were isolated. The possible role of these fungi in the physiology of silver fir seed germination process was discussed.     

Aureonitols A and B,Two New C13‐Polyketides from Chaetomium globosum,an Endophytic Fungus in Salvia miltiorrhiza

Two new C₁₃‐polyketides, aureonitols A and B ( 1 and 2 ), along with five known compounds ( 3 – 7 ), were isolated from the solid fermentation culture of the plant endophytic fungus Chaetomium globosum from the aerial parts of Salvia miltiorrhiza. The structures and absolute configurations of 1 and 2 were determined by comprehensive spectroscopic data analysis and computed methods. Compound 5 was found to display the remarkable antimicrobial activities against four multidrug‐resistant bacteria (Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, and Staphylococcus epidermidis) with MIC values of 3.13–6.25 μg/mL (ciprofloxacin: 0.78–1.56 μg/mL), and also against all tested fungal strains with MIC values of 3.13–25 μg/mL (ketoconazole: 0.78–12.50 μg/mL).     

Tailoring of recombinant FDH: effect of histidine tag location on solubility and catalytic properties of Chaetomium thermophilum formate dehydrogenase (CtFDH)

Abstract

Several protein expression systems can be used to get enzymes in required quantities and study their functions. Incorporating a polyhistidine tag is a beneficial way of getting various enzymes such as FDHs for industrial applications. The NAD⁺ dependent formate dehydrogenase from Chaetomium thermophilum (CtFDH) can be utilized for interconversion of formate to carbon dioxide coupled with the conversion of NAD⁺ to NADH. In this study, N-terminal His tagged CtFDH (N-CtFDH) and C-terminal His tagged CtFDH (C-CtFDH) was constructed to learn the effect of His tag location on the activity and kinetic parameters of the enzyme. The solubility of proteins was not affected by tag position, however, an interference on the N-terminal region caused a deterioration in specific activity and the kinetic ability of enzyme. The obtained results indicated that the C-terminus of the enzyme is an appropriate region for tag engineering. The C-CtFDH has an approximately three-fold larger specific activity and two-fold higher catalytic efficiency than N-CtFDH. The results suggest that insertion of a His-tag at the N-terminal or C-terminal end of CtFDH has different effects on the protein and the N-terminal fragment of the protein is crucial for the function of CtFDH.     

Evolution of multicopper oxidase genes in coprophilous and non-coprophilous members of the order sordariales

Multicopper oxidases (MCO) catalyze the biological oxidation of various aromatic substrates and have been identified in plants, insects, bacteria, and wood rotting fungi. In nature, they are involved in biodegradation of biopolymers such as lignin and humic compounds, but have also been tested for various industrial applications. In fungi, MCOs have been shown to play important roles during their life cycles, such as in fruiting body formation, pigment formation and pathogenicity. Coprophilous fungi, which grow on the dung of herbivores, appear to encode an unexpectedly high number of enzymes capable of at least partly degrading lignin. This study compared the MCO-coding capacity of the coprophilous filamentous ascomycetes Podospora anserina and Sordaria macrospora with closely related non-coprophilous members of the order Sordariales. An increase of MCO genes in coprophilic members of the Sordariales most probably occurred by gene duplication and horizontal gene transfer events.     

Has the prehistoric ice-man contributed to the preservation of living fungal spores?

Abstract There are arguments against the conclusion drawn by Haselwandter and Ebner that fungal spores have survived for some 5300 years on hay padding in the leather boots of frozen body discovered in the Austrian Alps. According to cryobiological experience, long-term survival of fungal spores is very unlikely at temperatures fluctuating between zero and −40°C. It is quite possible that living spores of these common species have recently reached this substratum.     

Screening microbial strain for improving the nutritional value of wheat and corn straws as animal feed

From 18 strains of cellulolytic microorganisms including bacteria and filamentous fungi, one strain of soft rot fungus identified as Chaetomium cellulolyticum was screened with respect to stronger decomposition ability of cellulose and hemicellulose and its ability for protein synthesis. As it grew on raw corn straw in solid layer fermentation (SLF) for 5 days, the amino acid content in the fermentation product attained 19.29% (w/w) from 6.43% while the total cell wall was reduced by 54%. A toxicity test with mice showed that the fermentation product is not poisonous. The two filamentous fungi, Trichoderma pseudokoningii S-38 and Penicillium decumbens JU-A10 produced large amounts of extracellular cellulase and hemicellulase in the SLF process, but their growth was limited and they sporulated profusely with regard to their value as animal feed products.     

High expression level of antioxidants and pharmaceutical bioactivities of endophytic fungus Chaetomium globosum JN711454

In order to maximize antioxidant activity of pharmaceutical bioactive endophytic fungus Chaetomium globosum JN711454 during fermentation process, designed fermentation experiments of culture media for three levels of eight culture factors were performed using a Taguchi orthogonal array (OA) design with layout L18 (2¹ × 3⁷). The agitation and the potato extract were the most significant affecting factors, and their interaction contributed significantly to fungus activity. The production of antioxidants was more favorable for static condition with 25 g potato extract/100 m. The remaining factors had no strong impact when considered individually. The validation of statistically optimized medium indicated the improvement of antioxidant activity to a level of twofold with approximately overall 40% enhancement in activity. The extract of optimized medium was investigated for various pharmaceutical bioactivities; it revealed a moderate antimicrobial activity, strong anticancer activity against HepG-2, UACC62 cell lines, an antiviral activity against HSV-2 virus, and strong inhibitory activity to butyrylcholinesterase enzyme, one of the neurohydrolase enzymes that play a major role in development of Alzheimer’s disease. As a result of applying statistical fermentation designs, the optimized conditions of endophytic fungus C. globosum JN711454 developed a cost-effective production medium by using inexpensive commercial potato extracts statically, which can lower the energy requirement and could become an efficient, economic, and viable fermentation process for production of pharmaceutical secondary metabolites.