Bioremediation of petroleum-contaminated saline soil by Acinetobacter baumannii and Talaromyces sp. and functional potential analysis using metagenomic sequencing.

Microbial remediation is a potential remediation method for petroleum-contaminated soil. In order to explore the petroleum degradation mechanism by microorganisms, the oilfield soil was remedied by Acinetobacter baumannii combined with Talaromyces sp. The degradation mechanism was studied by analyzing soil microbial community and functional genes through metagenomics during the degradation process. The result showed the degradation rate of petroleum was 65.6% after 28 days. The concentration of petroleum decreased from 1220 mg/kg to 420 mg/kg. In the co-culture group, Acinetobacter baumannii became the dominant species, the annotated genes of it at the species level accounted for 7.34% while that of Talaromyces sp. accounted for only 0.34%. Meanwhile, the annotated genes of Bacillus, Halomonas, and Nitriliruptor at the genus level were up-regulated by 1.83%, 0.90%, and 0.71%, respectively. In addition, large functional genes were significantly up-regulated, including the peroxisome, P450 enzyme (CYP53, CYP116, CYP102, CYP645), and biofilm formulation, promoting the oxidation and hydroxylation, and catalyzing the epoxidation of aromatic and aliphatic hydrocarbons. Meanwhile, the degrading genes of alkanes and aromatic hydrocarbons were expressed promotionally, and degradation pathways were deduced. In conclusion, the inoculation of Acinetobacter baumannii combined with Talaromyces sp. accelerated the degradation of petroleum in oilfield soil and improved the growth of indigenous petroleum-degrading bacteria. Many functional genes related to petroleum degradation were promoted significantly. These results proved the co-culture of bacteria-fungi consortium contributes to the bioremediation of petroleum-contaminated soil.

Talaromyces amestolkiae uses organic phosphate sources for the treatment of uranium-contaminated water.

Fungi have received particular attention in regards to alternatives for bioremediation of heavy metal contaminated locales. Enzymes produced by filamentous fungi, such as phosphatases, can precipitate heavy metal ions in contaminated environments, forming metal phosphates (insoluble). Thus, this research aimed to analyze fungi for uranium biomineralization capacity. For this, Gongronella butleri, Penicillium piscarium, Rhodotorula sinensis and Talaromyces amestolkiae were evaluated. Phytate and glycerol 2-phosphate were used as the phosphate sources in the culture media at pH 3.5 and 5.5, with and without uranium ions. After 4 weeks of fungal growth, evaluated fungi were able to produce high concentrations of phosphates in the media. T. amestolkiae was the best phosphate producer, using phytate as an organic source. During fungal growth, there was no change in pH level of the culture medium. After 3 weeks of T. amestolkiae growth in medium supplemented with phytate, there was a reduction between 20 and 30% of uranium concentrations, with high precipitation of uranium and phosphate on the fungal biomass. The fungi analyzed in this research can use the phytic acid present in the medium and produce high concentrations of phosphate; which, in the environment, can assist in the heavy metal biomineralization processes, even in acidic environments. Such metabolic capabilities of fungi can be useful in decontaminating uranium-contaminated environments.

New butyrolactone derivatives from the endophytic Fungus Talaromyces sp. CPCC 400783 of Reynoutria japonica Houtt.

Six new butyrolactone derivatives (1, 2a/2b, 3a/3b and 4), together with another two known derivatives (5 and 6) were isolated from the endophytic fungus Talaromyces sp. CPCC 400783. Their structures were established by a combination of spectroscopic analysis, including NMR and HRESIMS. The absolute configurations were elucidated by ECD experiments. Subsequently, compound 1, 3b, 4 and 5 exhibited good inhibitory effect against influenza A/WSN/33 (H1N1) virus with IC50 values of 21.93 ± 1.51, 21.54 ± 3.75, 18.36 ± 2.15 and 23.80 ± 3.05 µM respectively.

Induction of secondary metabolite production by fungal co-culture of Talaromyces pinophilus and Paraphaeosphaeria sp.

Fungal co-culture is a strategy to induce the production of secondary metabolites by activating cryptic genes. We discovered the production of a new compound, talarodone A (1), along with five known compounds 2-6 in co-culture of Talaromyces pinophilus and Paraphaeosphaeria sp. isolated from soil collected in Miyazaki Prefecture, Japan. Among them, the productions of penicidones C (2) and D (3) were enhanced 27- and sixfold, respectively, by the co-culture. The structure of 3 should be represented as a γ-pyridol form with the reported chemical shifts, but not as a γ-pyridone form, based on DFT calculation.

Atrorosins: a new subgroup of Monascus pigments from Talaromyces atroroseus.

A new series of azaphilone pigments named atrorosins have been isolated from the filamentous fungus Talaromyces atroroseus. Atrorosins have a similar azaphilone scaffold as the orange Monascus pigment PP-O, with a carboxylic acid group at C-1, but are unique by their incorporation of amino acids into the isochromene system. Despite that the atrorosin precursor PP-O, during fermentation, was initially produced as two isomers (3:2, cis:trans ratio), the atrorosins were surprisingly almost exclusively (99.5%) produced as the cis-form, possibly due to steric interactions with the incorporated amino acid. When grown on complex media, a whole range of atrorosins is produced, whereas individual atrorosins can be produced selectively during fermentation by supplementing with the desired primary amine-containing compound.

Talaromyolides A-D and Talaromytin: Polycyclic Meroterpenoids from the Fungus Talaromyces sp. CX11.

Talaromyolides A-D (1-4) and talaromytin (5) were isolated from a marine fungus Talaromyces sp. CX11. Their structures were unambiguously determined by nuclear magnetic resonance (NMR), mass spectrometry, X-ray crystallography experiments, and time-dependent density functional theory electronic circular dichroism calculations. Talaromyolides A and D represent two novel carbon skeletons. Talaromytin exhibits two slowly interconverting conformers in DMSO-d6 and CH3OH-d4 that were studied by temperature-dependent NMR experiments. Talaromyolide D exhibits potent antiviral activity against pseudorabies virus (PRV) with a CC50 value of 3.35 µM.

Withanolide production by fungal endophyte isolated from Withania somnifera.

Withanolides, the secondary metabolite from Withania species is used in Ayurvedic medicine and now proved to have potential use in treating cardiovascular, Alzheimer's disease, etc. Its production in plants varies between genotypes with very low yield. For improved industrial commercialisation, there is a need to increase its production. Endophytic fungi are symbiotically associated with plants and can synthesise the same bioactive compounds and natural products as their host plant. There are no reports available on withanolide-producing (endophytic) fungi. The present study identified an endophytic fungus (Taleromyces pinophilus) from leaves of Withania somnifera which produces withanolides in the medium. The structure of withanolide was confirmed by 1H NMR, LC-MS analyses and quantified by HPLC analysis. The fungus produces high amount of withanolide when compared to leaf and root of W. somnifera. The fungus can be exploited to produce the withanolide to meet its demand.

New Isocoumarins and Related Metabolites from Talaromyces flavus.

Two new isocoumarin derivatives, talaisocoumarins A (1) and B (2), and three new related metabolites, talaflavuols A-C (3-5) were isolated from the wetland soil-derived fungus Talaromyces flavus BYD07-13. Their structures were elucidated by spectroscopic (NMR) and MS analyses. The absolute configurations of 1 and 2 were determined by CD and an Rh2(OCOCF3)4-induced CD method. All compounds were evaluated for cytotoxic and antimicrobial activities. However, none of them showed any activity. The plausible biosynthetic pathways for 1-5 were also proposed.

In vitro cytotoxicity and antimicrobial activity of Talaromyces flavus SP5 inhabited in the marine sediment of Southern Coast of India.

Marine sediment samples were collected from the coastal areas of Southern India, particularly in Kanyakumari District. Twenty-eight different fungal strains were isolated. The screening of fungi from marine sediment was done to isolate a potent fungus that can produce bioactive compounds for biomedical applications. Only three strains viz Trichoderma gamsii SP4, Talaromyces flavus SP5 and Aspergillus oryzae SP6 were screened for further studies. The intracellular bioactive compounds were extracted using solvent extraction method. The crude extracts were tested for its anti-microbial and anti-cancer properties and analytically characterized using Gas Chromatography Mass Spectrometry (GC-MS). All the three extracts were active, but the extract from T. flavus SP5 was found to be more active against various human pathogens, viz., Pseudomonas aeruginosa ATCC 27853 (17.8 ± 0.1), Escherichia coli ATCC 52922 (18.3 ± 0.3), and Candida tropicalis ATCC 750 (17.7 ± 0.4). It also exhibited cytotoxic activity against HEp2 carcinoma cell line with the LC50 value of 25.7 µg·L-1. The GC-MS data revealed the presence of effective bioactive compounds. These results revealed that the extract from isolated fungus T. flavus SP5 acted as a potent antimicrobial, antifungal, and anticancer agent, providing basic information on the potency of marine fungi towards biomedical applications; further investigation may lead to the development of novel anticancer drugs.

Carbon and Nitrogen Sources Influence Tricalcium Phosphate Solubilization and Extracellular Phosphatase Activity by Talaromyces flavus.

The aim of this work was to study phosphate (P) solubilization (and the processes involved in this event) by Talaromyces flavus (BAFC 3125) as a function of carbon and/or nitrogen sources. P solubilization was evaluated in NBRIP media supplemented with different carbon (glucose, sorbitol, sucrose, and fructose) and nitrogen (L-asparagine, urea, ammonium sulfate (AS), and ammonium nitrate (AN) combinations. The highest P solubilization was related to the highest organic acid production (especially gluconic acid) and pH drop for those treatments where glucose was present. Also P solubilization was higher when an inorganic nitrogen source was supplemented to the media when compared to an organic one. Although not being present an organic P source, phosphatase activity was observed. This shows that P mineralization and P solubilization can occur simultaneously, and that P mineralization is not induced by the enzyme substrate. The combination that showed highest P solubilization was for AN-glucose. The highest acid phosphatase activity was for AS-fructose, while for alkaline phosphatase were for AS-fructose and AN-fructose. Acid phosphatase activity was higher than alkaline. P solubilization and phosphatase activity (acid and alkaline) were influenced by the different carbon-nitrogen combinations. A better understanding of phosphate-solubilizing fungi could bring a better use of soil P.

Characterization and flocculability of a novel proteoglycan produced by Talaromyces trachyspermus OU5.

A filamentous fungus strain OU5 was isolated from a soil sample for its ability to produce rich exopolymers (EPS), with high flocculation capability towards kaolin suspension and swine wastewater, at low-carbon source conditions. EPS from strain OU5 was extracted and characterized to determine its flocculating behavior and active constituents involved in the flocculation. Strain OU5 was identified as Talaromyces trachyspermus by 18S rDNA-ITS gene sequencing and morphological observation. The extracted EPS was a novel proteoglycan (designated as BF-OU5) composed of 84.6% (w/w) polysaccharides and 15.2% (w/w) proteins. The enzymatic digestion tests revealed that the polysaccharides in BF-OU5, composed of 67% glucose, 16.4% mannose, 8.6% xylose and 8% galactose, contributed to 99.7% of flocculating capacity and were the major active ingredients in the flocculation. By contrast, the proteins in BF-OU5 only had minor roles in the flocculation. The presence of hydroxyl, amide, carboxyl and methoxyl functional groups in BF-OU5, and the high molecular weight (1.053 × 10(5)-2.970 × 10(5) Da) as well as the structure of a spherical conformation with inner pores and channels made of cross-linked netted textures contributed to the flocculation. A dosage of 20 mg/l BF-OU5 initiated more than 92.5% of flocculating efficiency towards kaolin suspension without any added coagulants; its flocculability was stable over a wide range of pH (4.0-8.0) and temperature (20°C-100°C). Treatment of swine wastewater using BF-OU5 achieved 52.1% flocculating removal for chemical oxygen demand, 39.7% for Kjeldahl nitrogen, 18.6% for NH4(+)-N, 21.5% for total phosphorus, and 75% for turbidity.

Spiculisporic acid E, a new spiculisporic acid derivative and ergosterol derivatives from the marine-sponge associated fungus Talaromyces trachyspermus (KUFA 0021).

A new spiculisporic acid derivative, spiculisporic acid E (2), and a new natural product 3-acetyl ergosterol 5, 8-endoperoxide (1), were isolated, together with ergosta-4, 6, 8 (14), 22-tetraen-3-one, glaucanic acid and glauconic acid, from the culture of the marine-sponge associated fungus Talaromyces trachyspermus (KUFA 0021). All the compounds were inactive against Gram-positive and Gram-negative bacteria, and Candida albicans, as well as multidrug-resistant isolates from the environment. Spiculisporic acid E (2), glaucanic acid and glauconic acid did not show in vitro growth inhibitory activity against the MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer) and A375-C5 (melanoma) cell lines by the protein binding dye SRB method.

The bioactive metabolites of the mangrove endophytic fungus Talaromyces sp. ZH-154 isolated from Kandelia candel (L.) Druce.

Bioactivity-directed fractionation of the extract of the mangrove endophytic fungus Talaromyces sp. ZH-154, which was isolated from the stem bark of Kandelia candel (L.) Druce, Rhizophoraceae, afforded two new metabolites, 7-epiaustdiol ( 1) and 8-O-methylepiaustdiol ( 2), together with the known compounds, stemphyperylenol ( 3), skyrin ( 4), secalonic acid A ( 5), emodin ( 6), and norlichexanthone ( 7). Their structures were elucidated on the basis of spectroscopic evidences including CD, MS, and 1D, 2D NMR techniques. The absolute configuration of 1 was unequivocally determined by single-crystal X-ray diffraction. All isolated compounds were evaluated for their antimicrobial and in vitro cytotoxic activities.