• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.74-80
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• 1989

For improvement of photobiological hydrogen production, Rhodopseudomonas El5-1, a photo-synthetic becterium capable of producing n high yield of hydrogen, was immobilized and conditions for hydrogen production by immobilized cells were examined. The optimum concentration for the combined matrix was obtained when sodium alginate was used at final concentration of 4%. The immobilized cells may reduce the inhibitory effects of nitrogen or oxygen. To minimize the diffusion resistance of the nutrients in alginate gel, the bend size less than 2 mm in diameter was desirable. The immobilized cells were also able to utilize n wide range of organic substrates for the production of hydrogen. The hydrogen producing activity of the immobilized cells was maintained for 20 days without loss of activity during semi-continuous operation of the reactor by feeding of new medium periodically and continuous production of hydrogen could be successfully performed for 30 days.

• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.905-912
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• 2007

A novel ${\beta}-glucosidase$ gene, bglA, was isolated from uncultured soil bacteria and characterized. Using genomic libraries constructed from soil DNA, a gene encoding a protein that hydrolyzes a fluorogenic analog of cellulose, 4-methylumbelliferyl ${\beta}-D-cellobioside$ (MUC), was isolated using a microtiter plate assay. The gene, bglA, was sequenced using a shotgun approach, and expressed in E. coli. The deduced 55-kDa amino acid sequence for bglA showed a 56% identity with the family 1 glycosyl hydrolase Chloroflexus aurantiacus. BglA included two conserved family 1 glycosyl hydrolase regions. When using $p-nitrophenyl-{\beta}-D-glucoside$ (pNPG) as the substrate, the maximum activity of the purified ${\beta}-glucosidase$ exhibited at pH 6.5 and $55^{\circ}C$, and was enhanced in the presence of $Mn^{2+}$. The $K_m\;and\;V_{max}$ values for the purified enzyme with pNPG were 0.16 mM and $19.10{\mu}mol/min$, respectively. The purified BglA enzyme hydrolyzed both pNPG and $p-nitrophenyl-{\beta}-D-fucoside$. The enzyme also exhibited substantial glycosyl hydrolase activities with natural glycosyl substrates, such as sophorose, cellobiose, cellotriose, cellotetraose, and cellopentaose, yet low hydrolytic activities with gentiobiose, salicin, and arbutin. Moreover, BglA was able to convert the major ginsenoside $Rb_1$ into the pharmaceutically active minor ginsenoside Rd within 24 h.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.932-945
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• 2009

Several species of Gram-positive bacteria have cell wall peptidoglycan (syn. murein) in which not all of the sugar moieties are N-acetylated. This has recently been shown to be a secondary effect, caused by the action of a peptidoglycan N-acetylglucosamine deacetylase. We have found that the opportunistic pathogen Listeria monocytogenes is unusual in having three enzymes with such activity, two of which remain in the cytoplasm. Here, we examine the enzyme (PgdA) that crosses the cytoplasmic membrane and is localized in the cell wall. We purified a hexa-His-tagged form of PgdA to study its activity and constructed a mutant devoid of functional Lmo0415 (PgdA) protein. L. monocytogenes PgdA protein exhibited peptidoglycan N-acetylglucosamine deacetylase activity with natural substrates (peptidoglycan) from both L. monocytogenes and Escherichia coli as well as the peptidoglycan sugar chain component N-acetylglucosamine, but not with N-acetylmuramic acid. As was reported recently [6], inactivation of the structural gene was not lethal for L. monocytogenes nor did it affect growth rate or morphology of the cells. However, the pgdA mutant was more prone to autolysis induced by such agents as Triton X-100 and EDTA, and is more susceptible to the cationic antimicrobial peptides (CAMP) lysozyme and mutanolysin, using either peptidoglycan muramidases or autolysis-inducing agents. The pgdA mutant was also slightly more susceptible than the wild-type strain to the action of certain beta-lactam antibiotics. Our results indicate that protein PgdA plays a protective physiological role for listerial cells.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1782-1788
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• 2007

Cellulosomes in Clostridium cellulovorans are assembled by the interaction between the repeated cohesin domains of a scaffolding protein (CbpA) and the dockerin domain of enzyme components. In this study, we determined the synergistic effects on cellulosic and hemicellulosic substrates by three different recombinant mini-cellulosomes containing either endoglucanase EngB or endoxylanase XynA bound to mini-CbpA with one cohesin domain (mini-CbpAl), two cohesins (mini-CbpA12), or four cohesins (mini-CbpAl234). The assembly of EngB or XynA with mini-CbpA increased the activity against carboxymethyl cellulose, acid-swollen cellulose, Avicel, xylan, and com fiber 1.1-1.8-fold compared with that for the corresponding enzyme alone. A most distinct improvement was shown with com fiber, a natural substrate containing xylan, arabinan, and cellulose. However, there was little difference in activity between the three different mini-cellulosomes when the cellulosomal enzyme concentration was held constant regardless of the copy number of cohesins in the cellulosome. A synergistic effect was observed when the enzyme concentration was increased to be proportional to the number of cohesins in the mini-cellulosome. The highest degree of synergy was observed with mini-CbpAl234 (1.8-fold) and then mini-CbpAl2 (1.3-fold), and the lowest synergy was observed with mini-CbpAl (1.2-fold) when Avicel was used as the substrate. As the copy number of cohesin was increased, there was more synergy. These results indicate that the clustering effect (physical enzyme proximity) of the enzyme within the mini-cellulosome is one of the important factors for efficient degradation of plant cell walls.

• Journal of fish pathology
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• v.20 no.1
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• pp.33-40
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• 2007

Biochemical characteristic of Streptococcus iniae and Streptococcus parauberis that are pathogens of streptococcosis of cultured flounder, Paralichthys olivaceus, in Jeju area was examined. The result of experiments on the grow according to temperatures showed that only S. parauberis grew at 10℃, the result of hemolysis test showed that only S. iniae bacteria showed β hemolysis. Only S. parauberis were positive in VP test and HIP test, both bacteria used α-D-glucose, D-mannose, D-psicose, D-trehalose, pyruvatic acid methyl ester, and glycerol as substrates. L-lactic acid was used only S. iniae bacteria, and β-methyl-D-glucosid was used only by S. parauberis. S. iniae exhibited acute infection patten, differently S. parauberis exhibited chronic infection patten in pathogenic test.

• Journal of the Korean Society of Environmental Restoration Technology
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• v.4 no.1
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• pp.57-66
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• 2001

The floating islands have been constructed for the water quality improvement and the biodiversity conservation in an disturbed aquatic ecosystem. We made floating islands consisted of a special float and substrates of coconut fibers implanted with four emergent macrophytes such as Phragmites australis, Zizania latifolia, Iris pseudoacorus, Typha angustifolia. Vegetation structure and plant growth were compared between on the floating islands and on ground in order to select suitable plants for the construction of floating islands. Emergent-macrophytic vegetation on the floating islands showed lower coverages and higher plant biodiversity due to natural introduction of various hydrophytes and hygrophytes. Shoot density was increased on floating islands except for Zizania latifolia. From the point of coverage and density of plants, Phragmites australis and Iris pseudoacorus were suitable for floating islands. Total biomass of emergent macrophytes was decreased on the floating islands. The belowground/aboveground biomass ratio of floating islands was higher than that of the ground. Out of planted macrophytes, Iris pseudoacorus with a high belowground/aboveground biomass ratio could be evaluated a suitable plant for the floating islands because a plenty of its root is profitable to adapt with the nutrient-limited environment of floating islands.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.804-810
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• 1999

There is a possibility that carvone, a monoterpene from spearmint (Mentha spicata), could induce the bph degradative pathway and genes in Alcaligenes eutrophus H850, which is a known Gram-negative PCB degrader with a broad substrate specificity that was thoroughly investigated with Arthrobacter sp. BIB, a Gram-positive PCB degrader. The strains BIB and H850 were unable to utilize and grow on the plant terpene [(R)-(-)-carvone] (50ppm) to be recognized as a sole carbon source. Nevertheless, the carvone did induce 2,3-dihydroxybiphenyl 1,2-dioxygenase (encoded by bphC) in the strain B lB, as observed by a resting cell assay that monitors accumulation of a yellow meta ring fission product from 4,4'-dichlorobiphenyl (DCBp). The monoterpene, however, did not appear to induce the meta cleavage pathway in the strain H850. Instead, an assumption was made that the strain might be using an alternative pathway, probably the ortho-cleavage pathway. A reverse transcription (RT)-PCR system, utilizing primers designed from a conserved region of the bphC gene of Arthrobacter sp. M5, was employed to verify the occurrence of the alternative pathway. A successful amplification (182bp) of mRNA transcribed from the N-terminal region of the bphC gene was accomplished in H850 cells induced by carvone (50ppm) as well as in biphenyl-growth cells. It is, therefore, likely that H850 possesses a specific PCB degradation pathway and hence a different substrate specificity compared with B1B. This study will contribute to an elucidation of the dynamic aspects of PCB bioremediation in terms of roles played by PCB degraders and plant terpenes as natural inducer substrates that are ubiquitous and environmentally compatible.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.1
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• pp.61-66
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• 2001

The aim of this study was to examine how plant terpenoids, as natural growth substrates or inducers, would affect the biodegradation of PCB congeners. Various PCB degraders that could grow on biphenyl and several terpenoids were tested for their PCB degradation capabilities. Degradation activities of the PCB congeners, 4,4-dichlorobiphenyl (4,4-DCBp) and 2,2-dichlorobiphenyl (2,2-DCBp), were initially monitored through a resting cell assay technique that could detect their degradation products. The PCB degraders, Pseudomonas ((S)-(-) limonene, p-cymene and $\alpha$-terpinene) whereas Arthrobacter sp. B1B could not grow on the terpenoids as a sole carbon source. The B1B strain grown on biphenyl exhibited good degradation activity for 4,4-DCBp and 2,2-DCBp, while the activity of strains P166 and T104 was about 25% that of the B1B strain, respectively. Concomitant GC analysis, however, demonstrated that strain T104, grown on (S)-(-) limonene, p-cymene and $\alpha$-terpinene, could degrade 4,4-DCBp up to 30%, equivalent to 50% of the biphenyl induction level. Moreover, strain T104 grown on (S)-(-) limonene, could also degrade 2,2-DCBp up to 30%. This indicates that terpenoids, widely distributed in nature, could be utilized as both growth and/or inducer substrate(s) for PCB biodegradation in the environment.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1133-1140
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• 2018

Pseudomonas fluorescens KLR101 was found to be capable of producing polyhydroxyalkanoate (PHA) using various sugars and fatty acids with carbon numbers ranging from 2 to 6. The PHA granules consisted mainly of a poly(3-hydroxybutyrate) homopolymer and/or poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer. Genomic DNA of P. fluorescens was fractionated and cloned into a lambda library, in which a 5.8-kb fragment that hybridized to a heterologous phaC probe from Ralstonia eutropha was identified. In vivo expression in Klebsiella aerogenes KC2671 (pUMS), restriction mapping, Southern hybridization experiments, and sequencing data revealed that PHA biosynthesis by P. fluorescens relied upon a polypeptide encoded by a 1,683-bp non-operonal ORF, which was preceded by a possible -24/-12 promoter and highly similar to DNA sequences of a gene encoding PHA synthase in the genus Pseudomonas. In vivo expression of the putative PHA synthase gene ($phaC_{Pf}$) in a recombinant Escherichia coli strain was investigated by using glucose and decanoate as substrates. E. coli (${phaC_{Pf}}^+$, pUMS) grown in medium containing glucose accumulated PHA granules consisting mainly of 3-hydroxybutyrate, whereas only a trace amount of 3-hydroxydecanoate was detected from an E. coli fadR mutant (${phaC_{Pf}}^+$) grown in medium containing decanoate. In vitro enzymatic assessment experiments showed that 3-hydroxybutyryl-CoA was efficiently used as a substrate of purified $PhaC_{Pf}$, suggesting that the putative PHA synthase of P. fluorescens utilizes mainly short-chain-length PHA precursors as a substrate.

• Journal of Bio-Environment Control
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• v.11 no.4
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• pp.199-204
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• 2002

To overcome a decrease of Korean ginseng production caused by successive cropping, we have tried to develop a nutrient culture system for Korean ginseng production. For determining the optimal substrate, mixture of sand and TKS-2 (S＋T), peatmoss (P), reused rockwool (RR), and granular rockwool (GR) were investigated. The overall physico-chemical properties of RR fell into the reported optimal range for the ginseng cultivation. However, bulk density of S＋T was a little higher than that of soil in Korean ginseng fields. The top fresh weight of the ginseng was high in RR and S＋T substrates. The root fresh and dry weights in the RR were remarkably greater than that in the conventional soil (CS) of Korean ginseng fields. In terms of ginseng quality, the vitamin C content of ginseng root in nutrient culture was higher than that in CS. However, the contents of crude saponin and total ginsenosides in ginseng between in the nutrient culture and in the soil culture did not show any significant differences.