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( Ichiro Kamei ),( Nana Tomitaka ),( Taichi Motoda ),( Yumi Yamasaki ) 한국미생물생명공학회 2022 Journal of microbiology and biotechnology Vol.32 No.2
Phlebia sp. MG-60 is the salt-tolerant, white-rot fungus which was isolated from a mangrove forest. This fungus expresses three kinds of manganese peroxidase (MGMnP) isozymes, MGMnP1, MGMnP2 and MGMnP3 in low nitrogen medium (LNM) or LNM containing NaCl. To date, there have been no reports on the biochemical salt-tolerance of these MnP isozymes due to the difficulty of purification. In present study, we established forced expression transformants of these three types of MnP isozymes. In addition, the fact that this fungus hardly produces native MnP in a high-nitrogen medium (HNM) was used to perform isozyme-selective expression and simple purification in HNM. The resulting MGMnPs showed high tolerance for NaCl compared with the MnP of Phanerochaete chrysosporium. It was worth noting that high concentration of NaCl (over 200 mM to 1200 mM) can enhance the activity of MGMnP1. Additionally, MGMnP1 showed relatively high thermo tolerance compared with other isozymes. MGMnPs may have evolved to adapt to chloride-rich environments, mangrove forest.
( Joy L. Harry-asobara ),( Ichiro Kamei ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 Journal of microbiology and biotechnology Vol.29 No.5
Bacterial strains that improve mycelial morphology and growth of white-rot fungi in liquid medium could enhance the impact of white-rot fungi towards lesser recovery of neonicotinoids when cocultured. This was demonstrated by the recovery of clothianidin and acetamiprid from cocultures of the white-rot fungus Phlebia brevispora strains with two mycelial-growth-promoting bacteria, Enterobacter sp. TN3W-14 and Pseudomonas sp. TN3W-8. Clothianidin recovery from cocultures of white-rot fungi and bacteria was over 40% lower than that from axenic microbial cultures and mixed-bacterial cultures. About 20% less acetamiprid was equally recovered from both TMIC33929+TN3W-14 cocultures and mixedbacterial cultures than from axenic fungal and bacterial cultures.
Ralstonia pickettii Enhance the DDT Biodegradation by Pleurotus eryngii
( Adi Setyo Purnomo ),( Diana Maulianawati ),( Ichiro Kamei ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 Journal of microbiology and biotechnology Vol.29 No.9
DDT is a hydrophobic organic pollutant, which can be bio-accumulated in nature and have adverse consequences on the physical condition of humans and animals. This study investigated the relationship between the white-rot fungus Pleurotus eryngii and biosurfactant-producing bacterium Ralstonia pickettii associated with the degradation of DDT. The effects of R. pickettii on fungal development were examined using in vitro confrontation assay on a potato dextrose agar (PDA) medium. R. pickettii culture was added to the P. eryngii culture at 1, 3, 5, 7, and 10 ml (1 ml ≈ 1.44 × 10<sup>13</sup> CFU). After 7 d incubation, about 43% of the initial DDT (12.5 μM) was degraded by the P. eryngii culture only. The augmentation of 7 ml of R. pickettii culture revealed a more highly optimized synergism with DDT degradation being approximately 78% and the ratio of optimization 1.06. According to the confrontational assay, R. pickettii promoted the growth of P. eryngii towards the bacterial colony, with no direct contact between the bacterial cells and mycelium (0.71 cm/day). DDD (1,1-dichloro-2,2-bis(4-chlorophenyl) ethane), DDE (1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene), and DDMU (1-chloro-2,2-bis(4-chlorophenyl) ethylene) were identified as metabolic products, indicating that the R. pickettii could enhance the DDT biodegradation by P. eryngii.
Culture Conditions for Mycelial Growth and Anti-Cancer Properties of Termitomyces
Tharavecharak Suphachai,D’Alessandro-Gabazza Corina N.,Toda Masaaki,Yasuma Taro,Tsuyama Taku,Kamei Ichiro,Gabazza Esteban C. 한국균학회 2023 Mycobiology Vol.51 No.2
Termitomyces sp. that grow in symbiosis with fungus-farming Termites have medicinal properties. However, they are rare in nature, and their artificial culture is challenging. The expression of AXL receptor tyrosine kinase and immune checkpoint molecules favor the growth of cancer cells. The study evaluated the optimal conditions for the artificial culture of Termitomyces and their inhibitory activity on AXL and immune checkpoint molecules in lung adenocarcinoma and melanoma cell lines. The culture of 45 strains of Termitomyces was compared. Five strains with marked growth rates were selected. Four of the selected strains form a single cluster by sequence analysis. The mycelium of 4 selected strains produces more fungal mass in potato dextrose broth than in a mixed media. The bark was the most appropriate solid substrate for Termitomyces mycelia culture. The mycelium of all five selected strains showed a higher growth rate under normal CO2 conditions. The culture broth, methanol, and ethyl acetate of one selected strain (T-120) inhibited the mRNA relative expression of AXL receptor tyrosine kinase and immune checkpoint molecules in cancer cell lines. Overall, these results suggest the potential usefulness of Termitomyces extracts as a co-adjuvant therapy in malignant diseases.