• KSBB Journal
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• v.8 no.4
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• pp.341-350
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• 1993

As an effort to develop an alternative transportation fuel, the production of methanol from methane gas was studied using the resting cells of an obligatory methanotroph, Methylosinus trichosporium OB3b. The reaction was carried out in high concentration phosphate buffer solutions with the flask-grown cells containing the exclusively cytoplasmic methane monooxygenase (sMMO) activity. The methanol accumulation rate was observed to be 79nmo1/mg·min during the initial 4.5hr. Phosphate-dependent inhibition was found for both sMMO and methanol dehydrogenase (MDH) activities, and the inhibition constants were 185mM and 42mM, respectively. The inhibition mode was noncompetitive. Methanol was found to be very inhibitory to the sMMO activity and the inhibition constant (noncompetitive) was 21mM when propylene was used as substrate. The sMO activity in the resting cells was declined very fast and the rate became very high during the methanol production. These results indicate that the use of M. trichosporium OB3b as a biocatalyst for the methanol production is heavily dependent on the stable maintenance of the whole-cell SMO activity as well as the effective alleviation of product inhibition.

• Journal of Life Science
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• v.13 no.2
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• pp.163-167
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• 2003

Identification of species within the toxin-producing genus Alexandrium is vital for biotoxin monitoring and mitigation decisions regarding shellfish industry. In particular, the discrimination of resting cysts of only A. tamarense from that of Alexandrium spp. is considerable important to fundamentally monitor and predict this species before vegetative cells occur in the nature. Fluorescent cTAM-F1 DNA probe was responsible to not only binding the activity of the vegetative cells in A. tamarense, but also to the resting cysts, which was treated with methanol after fixation and stained by primuline on the surface The location of fluorescence in cultured vegetative cells and resting cysts was almost at tile bottom of the nucleus. The optimal incubation temperature and time using in situ hybridization were 50-$54^{\circ}C$ and 40-60 min, respectively, to penetrate the DNA probe into cell.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.489-492
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• 2000

The eventual goal of this study is to elucidate roles of plant terpenoids (e.g., cymene, limonene and others) as natural substrates in the cometabolic biodegradation of PCBs and to develop an effective PCB bioremediation technology. The aim of this study was to examine how plant terpenoids, as natural 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. The PCB congener degradation activities were first monitored through resting cell assay technique that could detect degradation products of the substrate. The congener removal was also confirmed by concommitant GC analysis. The PCB degraders, Pseudononas sp. P166 and Caynebacterium sp. T104 were found to grow on both biphenyl and terpenoids ((S)-(-) limonene, p-cymene and ${\alpha}-terpinene$) whereas Arthrobacter B1B could not grow on the terpenoids as a sole carbon source. The strain B1B grown on biphenyl showed a good degradation activity for 4,4'-dichlorobiphenyl (DCBp) while strains P166 and T104 gave about 25% of B1B activity. Induction of degradation by cymene, limonene and terpine was hardly detected by the resting cell assay technique. This appeared to be due to relatively lower induction effect of these terpenoids compared with biphenyl. However, a subsequent GC analysis showed that the congener could be removed up to 30% by the resting cells of T104 grown on the terpenoids. This indicates that terpenoids, widely distributed in nature, could be utilized as both growth and/or inducer substrate for PCB biodegradation.

• Microbiology and Biotechnology Letters
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• v.28 no.1
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• pp.33-38
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• 2000

In order to produce ascorbic acid-2-phosphate from ascorbic acid, bacteria capable of transforming ascorbic acid to ascorbic acid-2-phosphate were isolated from soils and the stock cultures in our laboratory. Among them, a newly isolated bacterium LSH-3 having the best ability of producing ascorbic acid-2-phosphate was selected and partially identified as Pseudomonas sp. The optimum conditions for the production of ascorbic acid-2-phosphate from ascorbic acid and using its resting cells as the source os enzyme were investigated. The results were summarized as follows: The optimum cultivation time and the cell weight for the production of ascorbic acid-2-phosphate was 14 hours and 100g/I(wet weight), respectively. And 0.1%(v/v) Trition X-100 was the most effective surfactant. The optimum concentrations of ascorbic acid and pyrophosphate were 400mM and 500mM, respectively, which led to produce 14.54g/I of ascorbic acid-2-phosphate. The most effective buffer was 50mM sodium acetate. The optimum pH and temperature were 4.5 and $40^{\circ}C$, respectively. Under the above conditions, 17.71 g/I of ascorbic acid-2-phosphate was produced from ascorbic acid after 32 hour-incubation, which corresponded to 17.5% of conversion rate based on ascorbic acid.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.769-773
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• 2007

With the aim to produce ascorbic acid-2-phosphate(AsA-2-P) from L-ascorbic acid(AsA, Vitamin C), nine bacteria conferring the ability to transform AsA to AsA-2-P were isolated from soil samples alongside known strains from culture collections. Most isolates were classified to the genus Brevundimonas by 16S phylogenetic analysis. Among them, Brevundimonas diminuta KACC 10306 was selected as the experimental strain because of its the highest productivity of AsA-2-P. The optimum set of conditions for the AsA-2-P production from AsA using resting cells as the source of the enzyme was also investigated. The optimum cultivation time was 16 h and the cell concentration was 120g/l(wet weight). The optimum concentrations of AsA and pyrophosphate were 550mM and 450mM, respectively. The most effective buffer was 50mM sodium formate. The optimum pH was 4.5 and temperature was $40^{\circ}C$. Under the above conditions, 27.5g/l of AsA-2-P was produced from AsA after 36 h of incubation, which corresponded to a 19.7% conversion efficiency based on the initial concentration of AsA.

• Food Science and Biotechnology
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• v.17 no.6
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• pp.1322-1326
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• 2008

Tannase catalyzes the hydrolysis of gallic acid esters and hydrolysable tannins. Twenty-two Lactobacillus strains with tannase activity were isolated from 7 types of kimchi. A polymerase chain reaction-based assay targeting the recA gene assigned all isolates to either Lactobacillus plantarum or Lactobacillus pentosus. The tannase activities of isolates measured in whole cells and cell-free extracts varied even within each species. The activities of the isolates varied with the assay method, but both methods indicated that isolate LT7 (identified as L. pentosus) showed the highest activity. The results of thin layer chromatography and high performance liquid chromatography, respectively, showed that tannic acid and gallic acid degraded to pyrogallol in resting L. pentosus LT7 cells. Therefore, the putative biochemical pathway for the degradation of tannic acid by L. pentosus implies that tannic acid is hydrolyzed to gallic acid and glucose, with the formed gallic acid being decarboxylated to pyrogallol. This study revealed the possible production of pyrogallol from tannic acid by the resting cell reaction with L. pentosus LT7.

• Molecules and Cells
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• v.26 no.6
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• pp.517-529
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• 2008

Porosomes are supramolecular, lipoprotein structures at the cell plasma membrane, where membrane-bound secretory vesicles transiently dock and fuse to release inravesicular contents to the outside during cell secretion. The mouth of the porosome opening to the outside, range in size from 150 nm in diameter in acinar cells of the exocrine pancreas, to 12 nm in neurons, which dilates during cell secretion, returning to its resting size following completion of the process. In the past decade, the composition of the porosome, its structure and dynamics at nm resolution and in real time, and its functional reconstitution into artificial lipid membrane, have all been elucidated. In this mini review, the discovery of the porosome, its structure, function, isolation, chemistry, and reconstitution into lipid membrane, the molecular mechanism of secretory vesicle swelling and fusion at the base of porosomes, and how this new information provides a paradigm shift in our understanding of cell secretion, is discussed.

• Korean Journal of Microbiology
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• v.31 no.2
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• pp.129-134
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• 1993

The pchAB genes of Pseudomonas sp. DJ-12 produce the enzymes of 4-chlorobipheny] (4CB) dioxygenase and dihydrodiol dehydrogenase which act on the first and second steps in degradation of 4CB and biphenyl. The genes were cloned in E coli XLI-Blue. The pcbAB genes of about 2.2 kb in size were contained in the pCUlO1 hybrid plasmid in the cloned cell of CUIOI. The genes were found to have their own promoter and three restriction sites for HindlII. 2,3-dihydroxybiphenyl was detected by the resting cell assay, as the metabolite transformed from biphenyl by the cloned cell of CUIOI. This means that the pcbAB genes are well expressed in E. coli. But dechlorination was unlikely involved in the pchAB gene expression but was believed to occur by functioning on 4CBA produced after ring-cleavage of 4CB.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.5
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• pp.471-479
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• 1999

The Kv channel activity in vascular smooth muscle cell plays an important role in the regulation of membrane potential and blood vessel tone. It was postulated that increased blood vessel tone in hypertension was associated with alteration of Kv channel and membrane potential. Therefore, using whole cell mode of patch-clamp technique, the membrane potential and the 4-AP-sensitive Kv current in cerebral arterial smooth muscle cells were compared between normotensive rat and one-kidney, one-clip Goldblatt hypertensive rat (lK,lC-GBH rat). Cell capacitance of hypertensive rat was similar to that of normotensive rat. Cell capacitance of normotensive rat and 1K,lC-GBH rat were $20.8{\pm}2.3$ and $19.5{\pm}1.4$ pF, respectively. The resting membrane potentials measured in current clamp mode from normotensive rat and 1K,lC-GBH rat were $-45.9{\pm}1.7$ and $-38.5{\pm}1.6$ mV, respectively. 4-AP (5 mM) caused the resting membrane potential hypopolarize but charybdotoxin $(0.1\;{\mu}M)$ did not cause any change of membrane potential. Component of 4-AP-sensitive Kv current was smaller in 1K,lC-GBH rat than in normotensive rat. The voltage dependence of steady-state activation and inactivation of Kv channel determined by using double-pulse protocol showed no significant difference. These results suggest that 4-AP-sensitive Kv channels playa major role in the regulation of membrane potential in cerebral arterial smooth muscle cells and alterations of 4-AP-sensitive Kv channels would contribute to hypopolarization of membrane potential in 1K,lC-GBH rat.

• Applied Biological Chemistry
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• v.37 no.6
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• pp.427-432
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• 1994

The effectiveness of the resting cells of a photosynthetic bacterium, Rhodocyclus gelatinosus KUP-74, was investigated on the production of extracellular ${\delta}-aminolevulinate$ (ALA). The ALA generating system with 1.05 mg cells per milliliter required 6 hr-incubation to obtain maximal yield of extracellular ALA. Under this condition ALA inducers, i.e., levulinate or L-glutamate showed relatively low effects to increase extracellular ALA production. Instead, the volume of the system and proper cell density appeared to be important factors for effective production of the ALA. The life-span of the resting cells was remarkably extended to maintain stable production of the ALA by immobilization of cells.