Penicillium Steckii Research Articles

Quantitative Monitoring of Cyclic Glycine-Proline in Marine Mangrove-Derived Fungal Metabolites.

This study developed and validated a robust UPLC-MS/MS method for quantifying cyclic glycine-proline (cGP) in mangrove-derived Penicillium and Aspergillus strains. The method demonstrated excellent linearity, precision, and recovery, with detection limits as low as 4.8 ng/mL. Penicillium pedernalense extract achieved a cGP content of 67.45 ± 1.11 ng/mL, with a corresponding fermentation yield of 29.31 ± 0.61 mg/L. This surpassed Penicillium steckii, which reached a content of 31.71 ± 0.31 ng/mL, with a yield of 8.51 ± 0.15 mg/L. This quantitative approach for metabolite analysis provides a viable method for screening these fungal strains, highlighting their potential for sustainable production of cyclic glycine-proline (cGP).

Antimicrobial Isochroman-Derived Atropo-Diastereomeric Dimers from Penicillium Steckii SCISO 41228.

A pair of unidentified atropo-diastereomeric dimers, penicisteckins G (1) and H (2), and twelve known compounds (3-16) were isolated from the marine coral-derived fungus Penicillium steckii SCISO 41228. Their structures including the absolute configuration were determined by HR-ESI-MS, ECD, 1D-, and 2D-NMR spectra. Compounds 1 and 2 exhibited moderate antibacterial activity against most tested pathogenic strains, especially for methicillin-resistant Staphyloccocus aureus (MASA) and Micrococcus luteus, with MIC values of 4.0 μg mL-1. In addition, compounds 2 and 3 exhibit potent radical scavenging activity against 2,2-diphenyl-1-picrylhydrazylhydrate (DPPH), with IC50 values of 10.76 and 8.66 μg mL-1, respectively.

Process optimization for the production of fibrinolytic enzyme by a novel marine fungus – Penicillium steckii KU1

The demand for microbial thrombolytic agents is alarmingly increasing, and the discovery of novel microbial species with significant level of fibrinolytic enzyme production are required to cope up with the current circumstances. Surprisingly, marine fungi remain largely unexplored in this regard, presenting an untapped potential for the production of fibrinolytic enzymes.A novel marine fungal strain (Penicillium steckii KU1) was isolated successfully from Kappil beach, Kerala. At the beginning Czapek-dox media was used for the production of fibrinolytic enzyme. Statistical optimization of the enzyme production was achieved initially by Plackett-Burman design (PB). Interactive effect of various process parameters was further scrutinized by Central Composite Design (CCD). Incubation time, inoculum size and concentration of casein were shown significant contribution towards production. Statistical optimization, resulted in a tenfold increment (454.49 ± 14.66 U/mL) [optimized conditions - 30 °C, pH 7, casein 2.7 % (w/v), inoculum 5.75 million spores/mL and 115 h incubation], against the predicted value 441.42 U/mL.

Arohynapene A Produced by <i>Penicillium steckii</i> JB-NW-2-1 Isolated from <i>Avicennia marina</i> (Forssk.) Vierh and Its Cytotoxic Activities

Mangrove-associated endophytic fungi are producers of secondary metabolites in unique and diverse structures with interesting biological activities such as antiviral, antifungal, antibacterial, anti-inflammatory, and cytotoxic agents. Endophytes play an important role in the physiological activities of the host plants, influencing the improvement of resistance to stress, insects, nematodes, and diseases. In this study, arohynapene A, a polyketide compound, was successfully isolated from the mangrove-derived fungus Penicillium steckii JB-NW-2-1 obtained from mangrove plant Avicennia marina (Forssk) Vierh. The structure was determined by a spectroscopic method including IR, MS, 1D-, and 2D-NMR techniques. This compound was evaluated for cytotoxic activities using resazurin assay against four cancer cells, HeLa cervical, MCF-7 breast cancer, B16-F10 melanoma, and A549 lung adenocarcinoma. The results showed no significant activities against all cancer cells tested (IC50 > 500 µM).

Identifying Marine-Derived Tanzawaic Acid Derivatives as Novel Inhibitors against Osteoclastogenesis and Osteoporosis via Downregulation of NF-κB and NFATc1 Activation.

To discover novel osteoclast-targeting antiosteoporosis leads from natural products, we identified 40 tanzawaic acid derivatives, including 22 new ones (1-8, 14-19, 27-32, 37, and 38), from the South China Sea mangrove-derived fungus Penicillium steckii SCSIO 41025. Penicisteck acid F (2), one of the new derivatives showing the most potent NF-κB inhibitory activity, remarkably inhibited osteoclast generation in vitro. Mechanistically, 2 reduced RANKL-induced IκBα degradation, NF-κB p65 nuclear translocation, the activation and nuclear translocation of NFATc1, and the relevant mRNA expression. NF-κB p65 could be a potential molecular target for 2, which has been further determined by the cellular thermal shift assay, surface plasmon resonance, and the gene knock-down assay. Moreover, 2 could also alleviate osteoporosis in ovariectomized mice by reducing the quantities of osteoclasts. Our finding offered a novel potential inhibitor of osteoclastogenesis and osteoporosis for further development of potent antiosteoporosis agents.

Antibacterial activities and molecular identification of endophytic fungi isolated from mangrove Avicennia marina

Abstract. Mulyani Y, Wulandari AP, Sinaga SE, Safriansyah W, Azhari A, Purbaya S, Abdullah FF, Farabi K, Shiono Y, Supratman U. 2023. Antibacterial activities and molecular identification of endophytic fungi isolated from mangrove Avicennia marina. Biodiversitas 24: 6923-6933. This research explored the antibacterial potential of endophytic fungi from the Avicennia marina's (Forssk.) Vierh leaves, stem bark, and root, located in Blanakan, Subang District, West Java. The screening process for antibacterial activity potential was conducted using the Kirby Bauer disk diffusion method. Subsequently, the molecular screening results of the most promising fungi species were identified using Internal Transcribed Spacer (ITS) markers. The results revealed the isolation of 30 fungal isolates. Among these, seven endophytic fungi exhibited significant antibacterial activity against Staphylococcus aureus ATCC 29213 and Vibrio harveyi ATCC 5339, with inhibition zones ranging from 7.88±1.52 to 23.60±0.77 mm. Through molecular identification, 7 endophytic fungi were identified, including Penicillium chrysogenum, Cladosporium anthropophilum, Trichosporon asahii, Cladosporium sphaerospermum, Fusarium verticillioides, Meyerozyma carpophila, and Penicillium steckii. Notably, C. anthropophilum exhibited the highest inhibition zones against S. aureus and V. harveyi measuring 23.60±0.77 and 21.80±0.26 mm, and showed (Minimum Inhibitory Concentration) MIC values of 15.625±0.98 and 31.25±0.39 ?g/mL, respectively. In this study, endophytic fungi isolated from different parts of A. marina exhibited promising antibacterial activity, with Cladosporium anthropophilum from the stem bark showing the highest potency against S. aureus and V. harveyi, suggesting their potential as a source of antibacterial agents. To the best of our knowledge, this is the inaugural study uncovering the isolation and antibacterial potential of endophytic fungi from the genera Cladosporium, Trichosporon, Fusarium, and Meyerozyma. These fungi were extracted from the bark, leaves, and roots of A. marina, situated within the unique mangrove ecosystem of Blanakan Sub-district, Subang District, West Java, Indonesia.

Endophytic fungus Penicillium steckii DF33 promoted tanshinones biosynthesis in Salvia miltiorrhiza by regulating the expression of CYP450 genes

Salvia miltiorrhiza, a prominent traditional Chinese medicinal resource, has been extensively employed in the management of cardiovascular and cerebrovascular ailments. Ensuring the consistency of S. miltiorrhiza raw materials revolves around the imperative task of maintaining stable tanshinones content and composition. An effective approach in this regard involves the utilization of endophytic fungi as inducers. Within this context, our study spotlights an endophytic fungus, Penicillium steckii DF33, isolated from the roots of S. miltiorrhiza. Remarkably, this fungus has demonstrated a significant capacity to boost the biosynthesis and accumulation of tanshinones. The primary objective of this investigation is to elucidate the underlying regulatory mechanism by which DF33 enhances and regulates the biosynthesis and accumulation of tanshinones. This is achieved through its influence on the differential expression of crucial CYP450 genes within the S. miltiorrhiza hairy roots system. The results revealed that the DF33 elicitor not only promotes the growth of hairy roots but also enhances the accumulation of tanshinones. Notably, the content of cryptotanshinone was reached 1.6452 ± 0.0925 mg g−1, a fourfold increase compared to the control group. Our qRT-PCR results further demonstrate that the DF33 elicitor significantly up-regulates the expression of most key enzyme genes (GGPPS, CPS1, KSL1, CYP76AH1, CYP76AH3, CYP76AK1, CYP71D411) involved in the tanshinone biosynthesis pathway. This effect is particularly pronounced in certain critical CYP450 genes and Tanshinone ⅡA synthase (SmTⅡAS), with their expression levels peaking at 7 days or 14 days, respectively. In summary, endophytic P. steckii DF33 primarily enhances tanshinone biosynthesis by elevating the expression levels of pivotal enzyme genes associated with the modification and transformation stages within the tanshinone biosynthesis pathway. These findings underscore the potential of employing plant probiotics, specifically endophytic and root-associated microbes, to facilitate the biosynthesis and transformation of vital constituents in medicinal plants, and this approach holds promise for enhancing the quality of traditional Chinese medicinal materials.

New Fusarin Derivatives from the Marine Algicolous Fungus Penicillium steckii SCSIO41040.

Five new fusarin derivatives, steckfusarins A-E (1-5), and two known natural products (6, 7), were isolated and identified from the marine algicolous fungus Penicillium steckii SCSIO 41040. The new compounds, including absolute configurations, were determined by spectroscopic analyses and calculated electronic circular dichroism (ECD). All new compounds were evaluated for their antioxidant, antibacterial, antifungal, antiviral, cytotoxic, anti-inflammatory, antioxidant, cholesterol-lowering, acetyl cholinesterase (AChE) enzyme and 6-phosphofructo-2-kinase (PFKFB3) and phosphatidylinositol-3-kinase (PI3K) inhibitory activities. The biological evaluation results revealed that compound 1 exhibited radical scavenging activity against 2,2-diphenyl-1-picrylhydrazylhydrate (DPPH), with an IC50 value of 74.5 µg/mL. In addition, compound 1 also showed weak anti-inflammatory activity at a concentration of 20 µM.

Tanzawaic Acid Derivatives from the Marine-Derived Penicillium steckii as Inhibitors of RANKL-Induced Osteoclastogenesis.

Seven new tanzawaic acid derivatives, steckwaic acids E-K (1-7), and one new benzene derivate (8), together with seven known tanzawaic acid analogues (9-16) were isolated from the marine algicolous fungus Penicillium steckii SCSIO 41040. The structures and absolute configurations of these new compounds (1-8) were determined by spectroscopic analyses, X-ray diffraction, and comparison of ECD spectra to calculations. Compounds 2, 10, and 15 inhibited lipopolysaccharide (LPS)-induced nuclear factor kappa-B (NF-κB) with IC50 values of 10.4, 18.6, and 15.2 μM, respectively. Compound 2 could suppress the receptor activator of NF-κB ligand (RANKL)-induced osteoclast differentiation in bone marrow macrophage cells (BMMCs). To the best of our knowledge, this is the first report of osteoclastogenesis inhibitory activity for tanzawaic acid derivatives.

Green synthesized TiO2 nanoparticles: Structural characterization and photoinduced antifungal activity against P. steckii

This study synthesized titanium dioxide (TiO2 ) nanoparticles (NPs) from mango leaf extract and investigated the features and antibacterial capabilities of three different. The microscopic morphological observation, scanning electron microscopy, and transmission electron microscopy results showed that all three NPs showed agglomeration phenomenon, and the TN-1 sample existed as large agglomerates, whereas the agglomeration phenomenon of TN-3 sample was improved by the modified, without large agglomerates. The biosynthetic TN-2 and TN-3 NPs were spherical and uniform in size, whereas those of the TN-3 sample was the smallest, ranging from 10 to 30nm. X-ray diffraction and Raman spectroscopy results exhibited that these were highly pure anatase NPs. The result of ultraviolet (UV)-visible-near-infrared spectral analysis showed that the TN-2 and TN-3 samples displayed higher UV absorption properties than the TN-1 samples and were highest in the modified NPs, which was more suitable for preparing chitosan-based nanocomposite material in future experiments and studies. The colony diameters of the TN-1, TN-2, and TN-3 treatment groups were 7.99, 7.80, and 6.86mm, respectively, after 120min of UV light induction at a wavelength of 365nm. Significant differences were evident between the TN-3 and the other two groups (p<0.05), indicating that the TN-3 sample more effectively inhibited Penicillium steckii than the other TiO2 NPs. PRACTICAL APPLICATION: Nanomaterials coated film preservation is widely used in fruit and vegetable preservation. In this paper, TiO2 nanomaterials will be green synthesized using mango leaf and structurally characterized, whereas antibacterial tests will be conducted against the mango fruit-specific bacterium Penicillium steckii, which will provide a theoretical basis for the storage and preservation of mango.

Identification of the polyketide synthase gene responsible for the synthesis of tanzawaic acids in Penicillium steckii IBWF104-06

Microorganisms have been used as biological control agents (BCAs) in agriculture for a long time, but their importance has increased dramatically over the last few years.The Penicillium steckii IBWF104-06 strain has presented strong BCA activity in greenhouse experiments performed against phytopathogenic fungi and oomycetes. P. steckii strains generally produce different antifungal tanzawaic acids; interesting compounds known to be catalyzed by polyketide synthetases in other fungi.Since the decalin structure is characteristic for tanzawaic acids, two polyketide synthase genes (PsPKS1 and PsPKS2) were selected for further analysis, which have similarity in sequence and gene cluster structure with genes that are known to be responsible for the biosynthesis of decalin-containing compounds.Subsequently, gene-inactivation mutants of both PsPKS1 and PsPKS2 have been generated. It was found, that the ΔPspks1 mutant cannot produce tanzawaic acids any more, whereas reintegration of the original PsPKS1 gene into the genome of ΔPspks1 reestablished tanzawaic acid production. The mutant ΔPspks2 is not altered in tanzawaic acids production.Interestingly, both mutants ΔPsPKS1 and ΔPsPKS2 still display strong BCA activity, indicating that the mechanism of action is not related to the production of tanzawaic acids.

Uncommon Polyketides from Penicillium steckii AS-324, a Marine Endozoic Fungus Isolated from Deep-Sea Coral in the Magellan Seamount

Four unusual steckwaic acids E–H (1–4), possessing a rarely described acrylic acid unit at C-4 (1–3) or a double bond between C-12 and C-13 (4) are reported for the first time, along with four new analogues (5–8) and two known congeners (9 and 10). They were purified from the organic extract of Penicillium steckii AS-324, an endozoic fungus obtained from a deep-sea coral Acanthogorgiidae sp., which was collected from the Magellan Seamount at a depth of 1458 m. Their structures were determined by the interpretation of NMR and mass spectroscopic data. The relative and absolute configurations were determined by NOESY correlations, X-ray crystallographic analysis, and ECD calculations. All compounds were tested for their antimicrobial activities against human- and aquatic-pathogenic bacteria and plant-related pathogenic fungi.

DNA metabarcoding of fungal communities in Heshouwu (Polygonum multiflorum Thunb.)

Heshouwu, derived from the dried root tuber of Polygonum multiflorum Thunb., is a widely used functional food and medicinal herb. Mycotoxin pollution of Heshouwu samples has become an emergent issue that cannot be ignored. In this study, the presence of fungi in commercial Heshouwu samples was investigated, and the differences in fungal community in samples from different collection sites and processing approaches were analyzed by DNA metabarcoding. Results showed that Ascomycota and Basidiomycota were the predominant phyla. At the genus level, Aspergillus (2.14%–94.05%), Cladosporium (0.22%–41.18%), Candida (0%–87.40%), Penicillium (0.01%–27.45%), and Alternaria (0%–32.29%) were the main genera. Meanwhile, six potential toxigenic fungi and one human pathogen, namely, Aspergillus flavus, Aspergillus nomius, Aspergillus tamarii, Fusarium solani, Penicillium paxillin, Penicillium steckii, and Candida tropicalis, were detected. In addition, differences in fungal diversity, community composition, trophic modes, and functional groups of Heshouwu samples from five collection sites and two processing approaches were observed. This study provides a comprehensive understanding of the fungal communities of Heshouwu, thereby providing a basis for future research on the prevention of mycotoxin contamination in this important medicinal herb.

Tanzawaic Acid Derivatives: Fungal Polyketides from the Deep-Sea Coral-Derived Endozoic Penicillium steckii AS-324

Ten new tanzawaic acids, namely, steckwaic acids A-D (1-4), 11-ketotanzawaic acid D (5), 6,15-dihydroxytanzawaic acid M (6), 15R-methoxytanzawaic acid M (7), 15S-methoxytanzawaic acid M (8), 8-hydroxytanzawaic acid M (9), and 8-hydroxytanzawaic acid B (10), together with four known analogues (11-14), were isolated and identified from the endozoic fungus Penicillium steckii AS-324, which was obtained from a deep-sea Acanthogorgiidae sp. coral that was collected from the Magellan seamount in the Western Pacific Ocean. Structurally, steckwaic acids A-D (1-4) represent the first tanzawaic acid derivatives containing a naphthalene ring. Their structures were elucidated by detailed interpretation of the NMR spectroscopic and mass spectrometric data. X-ray crystallographic analysis and ECD calculations confirmed their structures and absolute configurations. New compounds 1, 5, and 9 exhibited activities against aquatic pathogenic bacteria Micrococcus luteus, Vibrio anguillarum, and Pseudomonas aeruginosa with MICs of 2, 4, and 4 μg/mL, respectively.

Photo-Induced Antifungal Activity of Chitosan Composite Film Solution with Nano–Titanium Dioxide and Nano-Silver

This study was conducted to investigate the UV light-induced effect of chitosan-titanium dioxide-silver (CTS-TiO2-Ag) nanocomposite film solution against Penicillium steckii and the underlying physiological mechanism of this effect. With longer UV exposure time, pathogen inhibition increased. UV-photoinduced treatment for 120 min produced the smallest P. steckii colony diameter, at 4.85 mm. However, when this treatment was followed by 8 h of storage, the conductivity of the P. steckii culture medium reached its highest level, at 713 microsiemens per cm. After a 120-h growth period on mangoes under the same conditions, the lesion diameters and proportion of infected mangoes reached 12.61 mm and 41.67%, respectively. Because the P. steckii cell membrane was severely disrupted, its permeability increased, causing serious extravasation of intracellular protein and nucleic acid material. Malondialdehyde, catalase, and superoxide dismutase in the P. steckii cells reached maximum concentrations (2.1106 μmol/mL, 44.06 U/mL, and 24.67 U/mL, respectively) after 8 h of incubation. These results indicate significant P. steckii inhibition by the UV light induction of the CTS-TiO2-Ag nanocomposite film solution.

Identification of the Polyketide Synthase Gene Responsible for the Synthesis of Tanzawaic Acids in Penicillium Steckii Ibwf104-06

Identification of the Polyketide Synthase Gene Responsible for the Synthesis of Tanzawaic Acids in Penicillium Steckii Ibwf104-06

Penicisteckins A-F, Isochroman-Derived Atropisomeric Dimers from Penicillium steckii HNNU-5B18.

Penicisteckins A-D (1-4), two pairs of atropodiastereomeric biaryl-type hetero- and homodimeric bis-isochromans with 7,5'- and 7,7'-linkages and a pair of atropodiastereomeric 2-(isochroman-5-yl)-1,4-benzoquinone derivatives [penicisteckins E (5) and F (6)], were isolated from the Penicillium steckii HNNU-5B18. Their structures including the absolute configuration were determined by extensive spectroscopic and single-crystal X-ray diffraction analysis and TDDFT-ECD calculations. Both the bis-isochromans and the isochroman/1,4-benzoquinone conjugates represent novel biaryl scaffolds containing both central and axial chirality elements. The monomer anserinone B (8) exhibited potent antibacterial activities against Staphylococcus aureus ATCC 29213 and methicillin-resistant Staphylococcus aureus with minimal inhibition concentration values ranging from 2 to 8 μg mL-1. Plausible biosynthetic pathways of 1-6 are proposed, which suggest how the absolute configurations of the isolates were established during the biosynthetic scheme.

Diversified Polyketides and Nitrogenous Compounds from the Mangrove Endophytic Fungus Penicillium steckiiSCSIO 41025

Main observation and conclusionFour new polyketide decalin derivatives, penicisteck acids A—D (1—4), and three new nitrogenous compounds (7—9), together with eight previously reported compounds, were isolated from the mangrove endophytic fungus Penicillium steckii SCSIO 41025. Their structures and absolute configurations were characterized by analysis of comprehensive spectroscopic data combined with ECD calculations and modified Marfey's method. Structurally, compounds 1—3 were highly oxygenated decalin derivatives harboring an unusual propanoic acid unit at C‐1. Compound 8 was identified as a rare natural alkaloid‐N‐oxide confirmed by single crystal X‐ray diffraction analysis. A plausible biogenetic pathway of compounds 1—6 is proposed. Compounds 13 and 15 showed significant anti‐bacterial activity against Staphylococcus aureus with the MIC values of 3.9 and 7.8 μg/mL, respectively. Compounds 10 and 12 exhibited potent α‐glucosidase inhibitory activity with the IC50 values of 64.1 and 191.4 μmol/L, respectively.

Genomic and Chemical Investigation of Bioactive Secondary Metabolites From a Marine-Derived Fungus Penicillium steckii P2648.

Marine fungi of the genus Penicillium are rich resources of secondary metabolites, showing a variety of biological activities. Our anti-bacterial screening revealed that the crude extract from a coral-derived fungus Penicillium steckii P2648 showed strong activity against some pathogenic bacteria. Genome sequencing and mining uncovered that there are 28 secondary metabolite gene clusters in P2648, potentially involved in the biosynthesis of antibacterial compounds. Chemical isolation and structural determination suggested citrinin is the dominant component of the crude extracts of P2648, and our further tests confirmed that citrinin showed excellent activities against various pathogenic bacteria. Moreover, the gene cluster containing a homolog of the polyketide synthase CitS was identified as the citrinin biosynthesis gene cluster through genetic analysis. Interestingly, three isoquinoline alkaloids were unexpectedly activated and isolated from the Δcits mutant and structural determination by using high-resolution electron spray ionization mass spectroscopy (HRESIMS), 1D, and 2D NMR. Further antibacterial assays displayed that compounds 1 and 2, but not compound 3, showed moderate activities against two antibiotic-resistant pathogenic bacteria with minimum inhibitory concentration (MIC) of 16–32 μg/ml. In conclusion, our results demonstrated that citrinin and isoquinoline alkaloids represent as the major antibacterial agents in the coral-associated fungus P. steckii P2648, and our genomic and chemical analyses present evidence in support of P. steckii P2648 as a potent natural products source for anti-bacterial drug discovery.

Of fungi and ticks: Morphological and molecular characterization of fungal contaminants of a laboratory-reared Ixodes ricinus colony

Establishing and maintaining tick colonies in the laboratory is essential for studying their biology and pathogen transmission, or for the development of new tick control methods. Due to their requirement for very high humidity, these laboratory-bred colonies are frequently subject to fungal contamination. In the present study, we aimed to identify the fungal species that contaminated a laboratory-reared colony of Ixodes ricinus through microscopic observation and molecular identification. We identified three different taxa isolated from the ticks: Aspergillus parasiticus, Penicillium steckii, and Scopulariopsis brevicaulis. These three species are usually regarded as environmental saprophytic molds but both direct and indirect evidence suggest that they could also be considered as entomopathogenic fungi. Although we do not have any direct evidence that the fungi isolated from I. ricinus in this study could cause lethal infections in ticks, we observed that once infected, heavy fungal growth coupled with very high mortality rates suggest that studying the entomopathogenic potential of these fungi could be relevant to biological tick control.