- Dominance of Endospore-forming Bacteria on a Rotating Activated Bacillus Contactor Biofilm for Advanced Wastewater Treatment
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Seong Joo Park , Jerng Chang Yoon , Kwang-Soo Shin , Eung Ho Kim , Soobin Yim , Yeon-Je Cho , Gi Moon Sung , Dong-Geun Lee , Seung Bum Kim , Dong-Uk Lee , Sung-Hoon Woo , Ben Koopman
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J. Microbiol. 2007;45(2):113-121.
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DOI: https://doi.org/2525 [pii]
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The bacterial diversity inherent to the biofilm community structure of a modified rotating biological contactor wastewater treatment process, referred to as the Rotating Activated Bacillus Contactor (RABC) process, was characterized in this study, via both culture-dependent and culture-independent methods. On the basis of culture-dependent methods, Bacillus sp. were found to exist in large numbers on the biofilm (6.5% of the heterotrophic bacteria) and the microbial composition of the biofilms was quite simple. Only three phyla were identified-namely, the Proteobacteria, the Actinobacteria (High G+C Gram-positive bacteria), and the Firmicutes (Low G+C Gram-positive bacteria). The culture-independent partial 16S rDNA sequence analysis revealed a considerably more diverse microbial composition within the biofilms. A total of eight phyla were recovered in this case, three of which were major groups: the Firmicutes (43.9%), the Proteobacteria (28.6%), and the Bacteroidetes (17.6%). The remaining five phyla were minor groups: the Planctomycetes (4.4%), the Chlorobi (2.2%), the Actinobacteria (1.1%), the Nitrospirae (1.1%), and the Verrucomicrobia (1.1%). The two most abundant genera detected were the endospore-forming bacteria (31.8%), Clostridium and Bacillus, both of which are members of the Firmicutes phylum. This finding indicates that these endospore-forming bacteria successfully colonized and dominated the RABC process biofilms. Many of the colonies or clones recovered from the biofilms evidenced significantly high homology in the 16S rDNA sequences of bacteria stored in databases associated with advanced wastewater treatment capabilities, including nitrification and denitrification, phosphorus accumulation, the removal of volatile odors, and the removal of chlorohydrocarbons or heavy metals. The microbial community structures observed in the biofilms were found to correlate nicely with the enhanced performance of advanced wastewater treatment protocols.
- Purification and Characterization of Manganese Peroxidase of the White-Rot Fungus Irpex lacteus
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Kwang-Soo Shin , Young Hwan Kim , Jong-Soon Lim
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J. Microbiol. 2005;43(6):503-509.
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DOI: https://doi.org/2298 [pii]
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The production of manganese peroxidase (MnP) by Irpex lacteus, purified to electrophoretic homogeneity by acetone precipitation, HiPrep Q and HiPrep Sephacryl S-200 chromatography, was shown to correlate with the decolorization of textile industry wastewater. The MnP was purified 11.0-fold, with an overall yield of 24.3%. The molecular mass of the native enzyme, as determined by gel filtration chromatography, was about 53 kDa. The enzyme was shown to have a molecular mass of 53.2 and 38.3 kDa on SDS-PAGE and MALDI-TOF mass spectrometry, respectively, and an isoelectric point of about 3.7. The enzyme was optimally active at pH 6.0 and between 30 and 40oC. The enzyme efficiently catalyzed the decolorization of various artificial dyes and oxidized Mn (II) to Mn (III) in the presence of H2O2. The absorption spectrum of the enzyme exhibited maxima at 407, 500, and 640 nm. The amino acid sequence of the three tryptic peptides was analyzed by ESI Q-TOF MS/MS spectrometry, and showed low similarity to those of the extracellular peroxidases of other white-rot basidiomycetes.
- The Role of Enzymes Produced by White-Rot Fungus Irpex lacteus in the Decolorization of the Textile Industry Effluent
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Kwang-Soo Shin
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J. Microbiol. 2004;42(1):37-41.
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DOI: https://doi.org/2003 [pii]
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The textile industry wastewater has been decolorized efficiently by the white rot fungus, Irpex lacteus, without adding any chemicals. The degree of the decolorization of the dye effluent by shaking or stationary cultures is 59 and 93%, respectively, on the 8th day. The higher level of manganese-dependent peroxidase (MnP) and non-specific peroxidase (NsP) was detected in stationary cultures than in the cultures shaken. Laccase activities were equivalent in both cultures and its level was not affected significantly by the culture duration. Neither lignin peroxidase (LiP) nor Remazol Brilliant Blue R oxidase (RBBR ox) was detected in both cultures. The absorbance of the dye effluent was significantly decreased by the stationary culture filtrate of 7 days in the absence of Mn (II) and veratryl alcohol. In the stationary culture filtrate, three or more additional peroxidase bands were detected by the zymogram analysis.
- Purification and Characterization of Extracellular Temperature-Stable Serine Protease from Aeromonas hydrophila
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Soo-Jin Cho , Jong-Ho Park , Seong Joo Park , Jong-Soon Lim , Eung Ho Kim , Yeon-Jae Cho , Kwang-Soo Shin
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J. Microbiol. 2003;41(3):207-211.
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Extracellular protease, from Aeromonas hydrophila Ni 39, was purified 16.7-fold to electrophoretic homogeneity with an overall yield of 19.9%, through a purification procedure of acetone precipitation, and Q Sepharose and Sephacryl S-200 chromatographies. The isoelectric point of the enzyme was 6.0 and the molecular mass, as determined by Sephacryl S-200 HR chromatography, was found to be about 102 kDa. SDS/PAGE revealed that the enzyme consisted of two subunits, with molecular masses of 65.9 kDa. Under standard assay conditions, the apparent K_m value of the enzyme toward casein was 0.32 mg/ml. About 90% of the proteolytic activity remained after heating at 60 ℃ for 30 min. The highest rate of azocasein hydrolysis for the enzyme was reached at 60℃, and the optimum pH of the enzyme was 9.0. The enzyme was inhibited by the serine protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), by about 87.9%, but not by E64, EDTA, pepstatin or 1,10-phenanthroline. The enzyme activity was inhibited slightly by Ca_2^+, Mg_2^+ and Zn_2^+ ions.
- Characterization of the Proteolytic Activity of Bacteria Isolated from a Rotating Biological Contactor
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In Jae Park , Jerng Chang Yoon , Seong Joo Park , Eung Ho Kim , Yeon-Jae Cho , Kwang-Soo Shin
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J. Microbiol. 2003;41(2):73-77.
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Four proteolytic bacteria were isolated and identified from a rotating biological contactor based on Bacillus. The four isolates, Ni 26, 36, 39 and 49 were identified as B. vallismortis, B. subtilis, Aeromonas hydrophila and B. amyloliquefaciens, respectively, based on their biochemical properties and 16S rDNA sequence analyses. The optimal proteolytic activity was observed in the temperature and pH ranges of 40-70ºC and 8.0-8.5, respectively. The proteolytic activities of all the isolates were partially inhibited by phenylmethylsulfonylfluoride (PMSF), and the isolates Ni 26, Ni 39 and Ni 49 were inhibited by the metalloprotease inhibitor, 1,10-phenanthroline. Zymographic analyses of the culture supernatants revealed the presence of at least two proteases in all isolates.
- Cytokine-Inducing and T Cell Mitogenic Effects of Cordyceps hepialidicola
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Jong-Soon Lim , Seung-Hyung Kim , Jeong-Youl Choi , Jin-Seo Park , Seong Joo Park , Kwang-Soo Shin
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J. Microbiol. 2001;39(3):181-185.
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The morphological characteristics of newly isolated Cordyceps hepialidicola were characterized, and the phylogenetic relationships with other Cordyceps species were investigated using a sequence analysis of the internal transcribed spacer (ITS). The PCR product of 592 bp showed a homology of 92 and 91% with C. militaris and C. nutans, respectively. In an in vitro model using mouse peripheral blood mononuclear cells (PBMC), a methanol extract of C. hepialidicola induced multiple cytokines, including IFN-[gamma], IL-4, and IL-18. The extract also enhanced the percentages of the CD4^+ and CD8^+ T cells in the healthy murine PBMCs to 56.1% and 13.0%, respectively. The percentages of CD4^+ and CD8^+ in the untreated controls were 28.4 and 7.3%, and concanavalin A-treated positive controls were 62.4 and 18.3%, respectively.
- Anti-tumor Activity of the Fruitbody Extract of Basidiomycete, Phellinus linteus
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Jong-Soon Lim , Seung-Hyung Kim , Jin-Seo Park , Jeong-Youl Choi , Seong Joo Park , Kwang-Soo Shin
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J. Microbiol. 2001;39(2):121-125.
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Methanol extract prepared from the fruitbody of Phellinus linteus (EPL) showed anti-tumor and immuno-stimulating activities. The invasion activity of B16-F10 melanoma cells through a reconstituted basement membrane to the collagen-coated lower surface of the filters was inhibited about 67% by EPL (100 ug/ml). Also, EPL inhibited the expression of the mRNA for MMP-2 and MMP-9. In vivo treatment of C57BL/6 mice (150 mg/kg) with EPL for 14 days, the pulmonary colonization was found to be inhibited about 75%. Using reverse transcriptionpolymerase chain reaction (RT-PCR) analysis, we found that cytokine IL-12 and INF-[gamma] genes were induced by EPL. Furthermore, EPL stimulated the proliferation of CD4^+ (33.5%) and CD8^+ (17.7%) in mouse splenocytes.
- Isolation of a Medium Chain Length Polyhydroxyalkanoic Acids Degrading Bacterium, Janthinobacterium lividum
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Jin-Seo Park , Jeong-Youl Choi , Pil-Mun Joung , Seong Joo Park , Young Ha Rhe e , Kwang-Soo Shin
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J. Microbiol. 2001;39(2):139-141.
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Medium-chain length polyhydroxyalkanoic acids (MCL-PHAs) degrading bacterium was isolated from the soil. The bacterium was identified as Janthinobacterium lividum by its biochemical properties, cell membrane fatty acids composition, and 16S rDNA sequence analysis. The bacterium showed a similarity of 0.911 with J. lividum according to the cell membrane fatty acids analysis and a similarity of 97% in the 16S rDNA sequence analysis. Culture supernatant of the bacterium showed the highest depolymerase activity toward polyhydroxynonanoic acid (PHN) that did not degrade the poly-[beta]-hydroxybutyric acid (PHB). The esterase activity was also detected with p-nitrophenyl (PNP) esters of fatty acids such as PNP-octanoic acid, PNP-dodecanoic acid, PNP-decanoic acid, and PNP-hexanoic acid.
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