• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.31-37
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• 2020

We previously identified a new heparinase inhibitor fungal metabolite, named CRM646-A, which showed inhibition of heparinase and telomerase activities in an in vitro enzyme assay and antimetastatic activity in a cell-based assay. In this study, we elucidated the mechanism by which CRM646-A rapidly induced nucleus condensation, plasma membrane disruption and morphological changes by increasing intracellular Ca²⁺ levels. Furthermore, PD98059, a mitogen-activated protein kinase (MEK) inhibitor, inhibited CRM646-A-induced nucleus condensation through ERK1/2 activation in rat 3Y1 fibroblast cells. We identified CRM646-A as a Ca²⁺ ionophore-like agent with a distinctly different chemical structure from that of previously reported Ca²⁺ ionophores. These results indicate that CRM646-A has the potential to be used as a new and effective antimetastatic drug.

• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.803-813
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• 2015

The growth of Irpex lacteus F17 and manganese peroxidase (MnP) production in a selfdesigned tray bioreactor, operating in solid-state conditions at a laboratory scale, were studied. The bioreactor was divided into three layers by three perforated trays. Agroindustrial residues were used both as the carrier of bound mycelia and as a nutrient medium for the growth of I. lacteus F17. The maximum biomass production in the bioreactor was detected at 60 h of fermentation, which was consistent with the CO₂ releasing rate by the fungus. During the stationary phase of fungal growth, the maximum MnP activity was observed, reaching 950 U/l at 84 h. Scanning electron microscopy images clearly showed the growth situation of mycelia on the support matrix. Furthermore, the MnP produced by I. lacteus F17 in the bioreactor was isolated and purified, and the internal peptide sequences were also identified with mass spectrometry. The optimal activity of the enzyme was detected at pH 7 and 25℃, with a long half-life time of 9 days. In addition, the MnP exhibited significant stability within a broad pH range of 4-7 and at temperature up to 55℃. Besides this, the MnP showed the ability to decolorize the polymeric model dye Poly R-478 in vitro.

• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.988-998
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• 2015

The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl₂-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30^oC. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.8
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• pp.3137-3140
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• 2015

Objective: To explore the immune reconstitution of $CD4^+T$ cells after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and its relationship with invasive fungal infection (IFI) in patients with hematological malignancies. Materials and Methods: Forty-seven patients with hematological malignancies undergoing Allo-HSCT in Binzhou Medical University Hospital from February, 2010 to October, 2014 were selected. At 1, 2 and 3 months after transplantation, the immune subpopulations and concentration of cytokines were assessed respectively using flow cytometry (FCM) and enzyme linked immunosorbent assay (ELISA). The incidence of IFI after transplantation and its correlation with immune reconstitution of $CD4^+T$ cells were investigated. Results: The number of $CD4^+T$ cells and immune subpopulations increased progressively after transplantation as time went on, but the subpopulation cell count 3 months after transplantation was still significantly lower than in the control group (p<0.01). In comparison to the control group, the levels of interleukin-6 (IL-6) and IL-10 after transplantation rose evidently (p<0.01), while that of transforming growth factor-${beta}$ (TGF-${beta}$) was decreased (p<0.01). There was no statistically significant difference level of interferon-${\gamma}$ (IFN-${\gamma}$) (p>0.05). The incidence of IFI was 19.2% (9/47), and multivariate logistic regression revealed that IFI might be related to Th17 cell count (p<0.05), instead of Th1, Th2 and Treg cell counts as well as IL-6, IL-10, TGF-${beta}$ and IFN-${\gamma}$ levels (p>0.05). Conclusions: After Allo-HSCT, the immune reconstitution of $CD4^+T$ cells is delayed and Th17 cell count decreases obviously, which may be related to occurrence of IFI.

• Weed & Turfgrass Science
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• v.6 no.2
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• pp.157-164
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• 2017

Functional microorganisms decompose various organic matter by enzyme activity and suppress plant disease caused by pathogen. This study was conducted to isolate and select functional microorganisms with protein or carbohydrate degradation activities and antagonistic activity against turfgrass fungal pathogens for eco-friendly turfgrass management in golf course from compost containing livestock manure of poultry or swine. Totally 68 isolates collected from livestock manure compost strains were isolated and tested for their activities of amylase, protease and lipase and antagonistic activities against Rhizoctonia solani AG2-2, R. solani AG1-1, and Sclerotinia homoeocarpa. Among the isolates, 34 strains were selected as functional microbes showing higher activities of amylase and protease. Three isolates of ASC-14, ASC-18, and ASC-35 among the 34 strains were selected as antifungal bacterial strains repressing the above 3 turfgrass fungal pathogens. Analysis results of 16s rRNA gene sequence and phylogenic cluster indicated that ASC-14 and ASC-18 belonged to Bacillus amyloliquefaciens, while ASC-35 was B. subtilis, respectively.

• Korean Journal of Food Science and Technology
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• v.15 no.4
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• pp.326-332
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• 1983

Acorn wine was manufactured experimentally with koji inoculated the strain producing acorn tannin hydrolyzing enzyme in order to apply fungal tannase to food processing. Starch value of several Korean acorns was found to be 72.84 and the acorns were worthy of use as a carbohydrate food. Mixed koji was prepared by combination of rice and acorn powder at a ratio of 50to 50 and inoculation of Aspergillus oryzae producing amylase and Aspergillus sp. AN-11 producing tannase into the mixture in order to hydrolyze efficiently acorn tannin inhibiting alcohol fermentation in the medium, and then the mixed koji was used as a suitable koji to manufacture acorn wine. Acorn wine brewed with medium of the acorn powder treated with water and cooked and the mixed koji prepared was superior about two times to that brewed with medium of untreated acorn powder and general koji with respect to the rate of alcohol production and sugar fermentation during the 1st and 2nd brewing.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.4
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• pp.789-799
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• 1998

Literatures on microorganisms of traditional Korean nuruk published since 1945 were reviewed in this paper. Traditional Korean nuruk consists of raw barley and various grains. Traditional Korean nuruk consists of unbolied raw barely and various grains. They are ground to paste and moistened, and then naturally inoculated by airborne microorganisms. Therefore, many kinds of microorganisms such as fungi, yeast, and bacteria grwo in nuruk. Since 1945, new 14 species of Aspergillus and 9 species of Penicillium have been identified from traditional Korean nuruk. Total number of fungal species identified so far is now up to 38 species among 12 different genus. Among newly isolated fungal species, Aspergillus penicilloides and Penicillium, expansum showed not only high production rate of acid and amylase but also extreme stability of the enzyme at room temperature for 3 months. As examples of newly isolated yeast species, there are 5 species of Candida, 4 species of Hansenula, 1 species of Pichia and 1 species of Schizosaccharomyces. Total number of yeast species isolated so far is up to 18 species from different 8 genus. Newly isolated bacteria, were Bacillus pumilus, Lactobacillus casei and Leuconostoc mesenteroides.

• Mycobiology
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• v.47 no.2
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• pp.230-241
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• 2019

The Great Sebkha of Oran is a closed depression located in northwestern of Algeria. Despite the ranking of this sebkha among the wetlands of global importance by Ramsar Convention in 2002, no studies on the fungal community in this area have been carried out. In our study, samples were collected from two different regions. The first region is characterized by halophilic vegetation and cereal crops and the second by a total absence of vegetation. The isolated strains were identified morphologically then by molecular analysis. The biotechnological interest of the strains was evaluated by testing their ability to grow at different concentration of NaCl and to produce extracellular enzymes (i.e., lipase, amylase, protease, and cellulase) on solid medium. The results showed that the soil of sebkha is alkaline, with the exception of the soil of cereal crops that is neutral, and extremely saline. In this work, the species Gymnoascus halophilus, Trichoderma gamsii, the two phytopathogenic fungi, Fusarium brachygibbosum and Penicillium allii, and the teleomorphic form of P. longicatenatum observed for the first time in this species, were isolated for the first time in Algeria. The halotolerance test revealed that the majority of the isolated are halotolerant. Wallemia sp. and two strains of G. halophilus are the only obligate halophilic strains. All strains are capable to secrete at least one of the four tested enzymes. The most interesting species presenting the highest enzymatic index were Aspergillus sp. strain A4, Chaetomium sp. strain H1, P. vinaceum, G. halophilus, Wallemia sp. and Ustilago cynodontis.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.10
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• pp.1548-1557
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• 2019

Objective: To investigate the effects of dietary supplementation with different levels and molecular weights of fungal ${\beta}$-glucan on productive performances, health, carcass traits and meat quality in broilers. Methods: Two hundred and ten of one-day-old chicks with equal sex were assigned to seven experimental groups in $2{\times}4$ factorial arrangement. These groups were supplemented with (0, 10, 30, and 60 ppm) of molecular weight 1-3, 1-6 ${\beta}$-glucan (low or high). High molecular weight ${\beta}$-glucan (H: 943 kDa) was obtained from Ophiocordyceps dipterigena BCC 2073, whereas H with ${\gamma}$-Irradiation treatment was performed to achieve low molecular weight ${\beta}$-glucan (L: 8 kDa). Results: There was no statistical significance in productive performances, apparent digestibility and interaction between fixed factors along 42 days of experiment (p>0.05). A higher caecal amylase activity was present in the group that received L, while there was a dramatic decrease in H and the control groups, respectively (p<0.05). The increase of supplemental dose increased caecal amylase activity (p<0.05). Immunomodulatory effects from L was revealed by the marked increase of phagocytic activity, relative weight of thymus and bursa of fabricius (p<0.05). Similarly, the additive dose at 30 ppm provided the same results, whereas the only significant difference with supplementation at 60 ppm was an increase in phagocytic activity (p<0.05). Interestingly, villi height of broilers fed L was higher than other groups (p<0.05). The treatments did not influence haematology, blood chemistry, antibody production level against vaccination, carcass traits and meat quality (p>0.05). Conclusion: The supplementation of L at 30 ppm was suggested to achieve benefits of immune modulation without adverse effects on other parameters.

• Journal of Microbiology and Biotechnology
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• v.32 no.4
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• pp.504-513
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• 2022

Chitin deacetylase (CDA) inhibitors were developed as novel antifungal agents because CDA participates in critical fungal physiological and metabolic processes and increases virulence in soil-borne fungal pathogens. However, few CDA inhibitors have been reported. In this study, 150 candidate CDA inhibitors were selected from the commercial Chemdiv compound library through structure-based virtual screening. The top-ranked 25 compounds were further evaluated for biological activity. The compound J075-4187 had an IC50 of 4.24 ± 0.16 µM for AnCDA. Molecular docking calculations predicted that compound J075-4187 binds to the amino acid residues, including active sites (H101, D48). Furthermore, compound J075-4187 inhibited food spoilage fungi and plant pathogenic fungi, with minimum inhibitory concentration (MIC) at 260 ㎍/ml and minimum fungicidal concentration (MFC) at 520 ㎍/ml. Therefore, compound J075-4187 is a good candidate for use in developing antifungal agents for fungi control.