• Journal of Microbiology
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• v.42 no.2
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• pp.156-159
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• 2004

In general, the salinity of the ocean is close to 3.5％ and marine vibrios possess the respiratory chain-linked Na$\^$+/ pump. The influence of sodium chloride and the proton conductor carbonylcyanide m-chlo-rophenylhydrazone (CCCP) on the production of extracellular proteases in a marine Vibrio strain was examined. At the concentration of 0.5 M, sodium chloride minimally inhibited the activity of extra-cellular proteases by approximately 16％, whereas at the same concentration, the producton of extra-cellular proteases was severely inhibited. On the other hand, the production of extracellular proteases was completely inhibited by the addition of 2 ${\mu}$M CCCP at pH 8.5, where the respiratory chain-linked Na$\^$+/ pump functions.

• Journal of Life Science
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• v.6 no.3
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• pp.198-203
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• 1996

A thermolabile alkaline phosphatase has been purified through steps of osmotic shock, ammonium sulfate salting-out, and DAEA-cellulose chromatography from the cultured broth of the marine Vibrio sp. M-96 strain. The optimal temperature for the enzyme activity was 35$\circ$C. The optimal pH was pH11.0, and the range of pHstability was pH10.4 to 12.0. Thermal inactivation occured within 6 mintes at 60$\circ$C. The enzyme was considerably inactivated by 0.1mM concentrations of Hg$^{2+}$, Ni$^{2+}$ and Zn$^{2+}$, whereas activated up to 234% by 1mM of Mn$^{2+}$. The activation energy and deactivation energy by the Arrhenius equation were 4.02 Kcal/mol and 9.098 Kcal/mol, respectively. The Km and Vmax values of the enzyme for p-introphenylphosphate were found to be 0.0465mM and 0.001334mM/min, respectively. Active form of the enzyme had a molecular weight of 57,000 dalton determined by the Sephadex G-200 gel filtration method.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.31-32
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• 2005

Neoagaro-oligosaccharides are produced only by enzymatic degradation of agarose by ${\beta}-agarase.^{1)}$ Neoagaro-oligosaccharides inhibit the growth of bacteria, slow the rate of degradation of starch, are used as low-calorie additives to improve food quality, and have macrophage-stimulating activity. Furthermore, neoagarobiose is a rare reagent that has both moisturizing effect on skin and whitening effect on melanoma $cells.^{2)}$ An agar-degrading marine bacterium was isolated from the sea water at the northeast coast in Cheju island, Korea. The strain was gram negative, aerobic, and motile rod. The 16S rRNA of the strain had the closest match of 98% homology, with that from Agarivorans albus. On the basis of several phenotypic characters and a phylogenetic analysis, this strain was designated Agarivorans sp. JA-1. In solid agar plate, Agarivorans sp. JA-1 produced a diffusible agarase that caused agar softening around the colonies. Agarivorans sp. JA-1 was cultured for 36 hr in marine broth 2216 (Difco, USA) and the supernatant that containing an extracellular ${\beta}-agarase$ was prepared by centrifugation of culture media. The enzyme exhibited relatively strong activity at $40^{\circ}C$ and was stable up to $60^{\circ}C$. Using PCR primers derived from the ${\beta}-agarase$ gene of Vibrio sp., the gene encoding ${\beta}-agarase$ from Agarivorans sp. JA-1 was cloned and sequenced. The structural gene consists of 2931 bp encoding 976 amino acids with a predicted molecular weight of 107,360 Da. The deduced amino acid sequence showed 99% and 34% homology to $agaA^{2)}$ and $agaB^{2)}$ genes for ${\beta}-agarase$ from Vibrio sp., respectively. The expression plasmid for ${\beta}-agarase$ gene of Agarivorans sp. JA-1 is being constructed and the recombinant enzyme will be biochemically characterized.

• Journal of Life Science
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• v.19 no.5
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• pp.566-572
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• 2009

A recombinant plasmid, pDH3, was obtained from the genomic library of Serattia marcescens KCTC 2172, and several recombinant subclones constructed from pDH3. The nucleotide sequence of a 5,137 bp segment, pPH4, was determined and three open reading frames were detected. The three ORFs encoded the phosphate specific transport (pst) operon, which was pstC, pstA, and pstB, with the same direction of transcription. Comparison of the pst operon of S. marcescens with that of other organisms revealed that the genes for pstS and phoU were missing. A potential CRP bonding site and pho box sequence was found in the upstream of the putative promoter at the regulatory region. Analysis of the nucleotide sequence showed that homology in amino acid sequences between the PstC protein and Yersinia sp., Vibrio sp., and Pseudomonas sp. were 49, 37 and 33%, respectively. The PstA protein and Yersinia sp., Vibrio sp., and Pseudomonas sp. showed homologies of 64, 51, and 47%, respectively. PstB protein and Methanocaldococcus sp., E. coli, and Mycoplasma sp. showed homologies of 60, 50, and 48%, respectively. The pst genes could be expressed in vivo and positively regulated by cAMP-CRP. The E. coli strain harboring plasmid pPH7, with pst genes, increased with the transport of phosphate.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.1
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• pp.89-95
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• 2013

Biological and functional characteristics of lactic acid bacteria (LAB) were investigated in mustard stem/leaf kimchi (MK), cabbage kimchi (CK), young radish kimchi (YRK), and cubed radish kimchi (CRK). LAB of young radish kimchi were mainly composed of bacilli in contrast to the other kimchi. 89.2% LAB isolated from all kimchi harbored plasmids. However, LAB had an average of $4.1{\pm}0.5$ plasmid bands in YRK, more than MK, CK, and CRK. Exopolysaccharides were produced by 10.9~11.1% of LAB, and were especially by LAB isolated from radish kimchi. A significant percentage of LAB (69.5%) had antibacterial activity against one sensitive strain or more. LAB from CK, YRK and CRK had antimicrobial activities against Bacillus sp., Listeria monocytogenes, and Salmonella Typhimurium, while the LAB from MK had activities against Vibrio parahaemolyticus higher than those from the other kimchi. In YRK and CRK, acid-tolerant LAB were twice as prevalent as those in MK and CK. Bile-tolerant LAB isolated from CRK were more prevalent than other kimchi. When $10^8$ CFU of LAB were added to Caco-2 cells, 12.1% of LAB isolated from all kimchi showed similar adherent activity to Lactobacillus rhamnosus GG. LAB of MK particularly adhered to Caco-2 cells, 2.0~4.1 fold higher than LAB in the other kimchi. From these results, biological and functional characteristics of LAB varied according to the type of kimchi and LAB existing in kimchi were limited to their respective species.