• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.616-623
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• 2008

In spite of an increasing interest in fucoidans as biologically active compounds, no convenient commercial sources with fucoidanase activity are yet available. A marine bacterial strain that showed confluent growth on a minimal medium containing fucoidan, prepared from Korean Undaria pinnatifida sporophylls, as the sole carbon source was isolated and identified based on a 16S rDNA sequence analysis as a strain of Sphingomonas paucimobilis, and named Sphingomonas paucimobilis PF-1. The strain depolymerized fucoidan into more than 7 distinct low-molecular-mass fucose-containing oligosaccharides, ranging from 305 to 3,749 Da. The enzyme activity was shown to be associated with the whole cell, suggesting the possibility of a surface display of the enzyme. However, a whole-cell enzyme preparation neither released the monomer L-fucose from the fucoidan nor hydrolyzed the chromogenic substrate p-nitrophenyl-${\alpha}$-L-fucoside, indicating that the enzyme may be an endo-acting fucoidanase rather than an ${\alpha}$-L-fucosidase. Therefore, this would appear to be the first report on fucoidanolytic activity by a Sphingomonas species and also the first report on the enzymatic degradation of the Korean Undaria pinnatifida sporophyll fucoidan. Moreover, this enzyme activity may be very useful for structural analyses of fucose-containing polysaccharides and the production of bioactive fucooligosaccharides.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.209-214
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• 2008

VBNC (Viable but nonculturable) state is an adaptive response of cells in adverse environments, which lead cell not grow on routine nutrient agar. In this study, we induced VBNC in Escherichia coli using copper and verify the characterization of it. After treatment of copper, we didn't detect any cells via plate cultivation, namely, colony forming unit (CFU) was zero. However, we identified the existence of VBNC by staining live cells with Live/Dead BacLight bacterial viability kit and counting them through flow cytometry. Then we isolated genomic DNA and RNA from VBNC-induced cells and analyzed the stability of them. Degradation of RNA is more severe than that of DNA and RNA is degraded as specific fragments. In addition, we showed the morphology of VBNC cell by Bio-Transmission Electron Microscope (Bio-TEM). VBNC cell showed impaired periplasmic space and inner and outer membrane were separated and the amount of cytosol were significantly decreased.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.109-111
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• 2010

Recent studies have described various microorganisms that can degrade PAH, however, there are currently limited methods available to screen for PAH-degrading microorganisms. To screen for PAH-degrading microorganisms, a sublimation method (Alley, Jeremy F. and Lewis R. Brown. 2000. Appl. Environ. Microbiol. 66, 439-442) was modified to produce a simple screening system. In our results, there were several bacterial species capable of pyrene degradation including genera, Coryenbacterium, Gordonia, Rhodococcus, and Streptomyces, which have been screened from 350 bacterial isolates of commercial gasoline and oil-spilled sediment by the sublimation method. The main advantage of this method is that it (i) safely deposits an even, thin and visible layer of PAH onto the agar surface without the use of solvents and (ii) the quantity of PAH sublimed onto the agar can be easily controlled. Overall, this sublimation method may be an effective and simple technique to screen for PAH-degrading microorganisms.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.382-390
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• 2006

Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

• 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.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.342-349
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• 2017

Polylactic acid (PLA) has been highlighted as an alternative renewable polymer for the replacement of petroleum-based plastic materials, and is considered to be biodegradable. On the other hand, the biodegradation of PLA by terminal degraders, such as microorganisms, requires a lengthy period in the natural environment, and its mechanism is not completely understood. PLA biodegradation studies have been conducted using mainly undefined mixed cultures, but only a few bacterial strains have been isolated and examined. For further characterization of PLA biodegradation, in this study, the PLA-degrading bacteria from digester sludge were isolated and identified using a polymer film-based screening method. The enrichment of sludge on PLA granules was conducted with the serial transference of a subculture into fresh media for 40 days, and the attached biofilm was inoculated on a PLA film on an agar plate. 3D optical microscopy showed that the isolates physically degraded the PLA film due to bacterial degradation. 16S rRNA gene sequencing identified the microbial colonies to be Pseudomonas sp. MYK1 and Bacillus sp. MYK2. The two isolates exhibited significantly higher specific gas production rates from PLA biodegradation compared with that of the initial sludge inoculum.

• Journal of Microbiology and Biotechnology
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• v.21 no.11
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• pp.1174-1178
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• 2011

Exopolysaccharides (EPSs) are microbial polysaccharides that are released outside of the bacterial cell wall. There have been few studies on EPS-producing lactic acid bacteria that can enhance macrophage activity and the underlying signaling mechanism for cytokine expression. In the current study, EPS-overproducing Lactobacillus (L.) paracasei KB28 was isolated from kimchi and cultivated in conditioned media containing glucose, sucrose, and lactose. The whole bacterial cells were obtained with their EPS being attached, and the cytokine-inducing activities of these cells were investigated. Gas chromatography analysis showed the presence of glucose, galactose, mannose, xylose, arabinose, and rhamnose in EPS composition. EPS-producing L. paracasei KB28 induced the expression of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-12 in mouse macrophages. This strain also caused the degradation of $I{\kappa}B{\alpha}$ and phosphorylation of the major MAPKs: Jun N-terminal kinase (JNK), p38, and extracellular signal-regulated kinase (ERK)1/2. The use of pharmacological inhibitors showed that different signaling pathways were involved in the induction of TNF-${\alpha}$, IL-6 and IL-12 by L. paracasei KB28. Our results provide information for a better understanding of the molecular mechanisms of the immunomodulatory effect of food-derived EPS-producing lactic acid bacteria.

• Korean Journal of Soil Science and Fertilizer
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• v.48 no.5
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• pp.485-491
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• 2015

This study was performed to investigate thermophilic bacteria from soil having broad antifungal spectrum against Rhizoctonia solani, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f.sp. lycopersici, and Botrytis cinerea. One isolate selected could resist heat shock of $60^{\circ}C$ for one hour, and had broad antifungal activity in dual culture assay against all tested fungal pathogens and was identified as Bacillus amyloliquefaciens Y1 using 16S rRNA gene sequence. Further investigation for antifungal activity of bacterial culture filtrate (BCF) and butanol crude extract (BCE) of various concentrations showed broad spectrum antifungal activity and fungal growth inhibition significantly increased with increasing concentration with highest growth inhibition of 100% against R. solani with 50% BCF and 11 mm of zone of inhibition against R. solani with 4 mg BCE concentration. Treatment of butanol crude extract resulted in deformation, lysis or degradation of C. gloeosporioides and P. capsici hyphae. Furthermore, B. amyloliquefaciens Y1 produced volatile compounds inhibiting growth of R. solani (70%), C. gloeosporioides (65%) and P. capsici (65-70%) when tested in volatile assay. The results from the study suggest that B. amyloliquefaciens Y1 could be a biocontrol candidate to control fungal diseases in crops.

• Advances in environmental research
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• v.2 no.3
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• pp.229-243
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• 2013

Coagulation-flocculation (CF) was tested coupled with an aerobic biofilter to reduce total petroleum hydrocarbon (TPHs) concentration and toxicity from petrochemical wastewater. The efficiency of the process was followed using turbidity and chemical oxygen demand (COD). The biofilter was packed with a basaltic waste (tezontle) and inoculated with a bacterial consortium. Toxicity test were carried out using Lactuca sativa var. capitata seeds. Best results for turbidity removal were obtained using alum. Considerable turbidity removal was obtained when using Opuntia spp. COD removal with alum was 25%, for Opuntia powder it was 36%. The application of the biofilter allowed the removal of 70% of the remaining TPHs after 30 days with a biodegradation rate (BDR) value 47 $mgL^{-1}d^{-1}$. COD removal was slightly higher with BDR value 63 $mgL^{-1}d^{-1}$. TPH kinetics allowed a degradation rate constant equal to $4.05{\times}10^{-2}d^{-1}$. COD removal showed similar trend with $k=4.23{\times}10^{-2}d^{-1}$. Toxicity reduction was also successfully achieved by the combined treatment process.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.321-326
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• 1989

Eighty two bacterial strains have been isolated from five different soil and sewage samples by selective enrichment culture on m-toluate minimal medium. Two of these were identified as Pseudomonas capacia, one as P. putida, one as Yersinia intermedia, and one as Flavobaeterium odoratum. P. cepacia SUB37 appeared to carry plasmid superficially similar to TOL plasmid previously described in p. putida mt-2 and other two plasmids from Flavobacterium odorutum and Y. intermedia larger than that of p. putida mt-2. p. cepacia SUB37 was sensitive to streptomycin but resistant to rifampicin. P. cepacia SUB37 carrying plasmid metabolizes the hydrocarbons to benzoate and toluates via the corresponding alcohols and aldehydes. By the curing experiment, it appears that P. cepacia SUB37 carries TOL plasmid encoding for the enzymes responsible for the catabolism of toluene and xylene via benzoate and the toluates and then by meta pathway in the process of degradation of aromatic hydrocarbons. p. cepacia SUB37 degraded m-toluate rapidly to be very low level when it was fully grown.