• Journal of Life Science
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• v.15 no.1 s.68
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• pp.94-99
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• 2005

Sixty four bacterial colonies which were able to oxidize the manganese were isolated from soil samples in Mokcheon and Ochang area. Among them, one bacterial strain was selected for this study based on its higher manganese oxidation, and this selected bacterial strain was identified as Aeromonas sp. MN44 through physiological-biochemical test and analysis of its 16s rRNA sequence. Aeromonas sp. MN44 was able to utilize lactose but did not utilize various carbohydrates as a sole carbon source. Aeromonas sp. MN44 showed a very sensitive to antibiotics such as kanamycin, chloramphenicol, ampicillin, tetracycline and spectinomycin, and heavy metal such as cadmium. But this strain showed a high resistance up to mg/ml unit to heavy metals such as lithium and manganese. Optimal manganese oxidation condition of Aeromonas sp. MN44 was pH 7.4 and manganese oxidation activity was inhibited by proteinase K and boiling treatment. So, we concluded that this factor was protein. The manganese oxidizing factor produced by Aeromonas sp. MN44 was partial purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 113 kDa.

• Korean Journal of Plant Resources
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• v.14 no.1
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• pp.65-70
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• 2001

This study was performed to select marker which can identify genetic variation between mother plant and in vitro cultured plantlets of strawberry by PCR using random primer. When 'Yeobong' DNA extracted was treated with proteinase-K and RNase-H, clear DNA bands were shown. The optimal condition for RAPD in strawberry was to use 50ng of template DNA, 10pmol of primer,37oC of annealing temperature, and 45 cycles of PCR. After establishing above PCR optimal condition, RAPD pattern was investigated by using UBC primers. PCR was performed, and 46 of 90 primers produced PCR product showing 158 total bands. GC content was compared between the primers forming bands and no bands. The GC content showing bands was average 67.4%, whereas primers showing no bands 58%.

• Journal of Dairy Science and Biotechnology
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• v.30 no.2
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• pp.119-129
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• 2012

Lactic acid bacteria (LAB) have been used as starter cultures in the manufacturing processes of fermented dairy products such as cheese and yogurt. LAB have a proteolytic system to use the nitrogen source from milk for their growth. The proteolytic system involved in casein utilization provides cells with essential amino acids during growth in milk and is also of industrial importance, because of its contribution to the development of the organoleptic properties such as flavor of fermented milk products. In the most extensively studied LAB, Lactococcus lactis, the main features of the proteolytic system comprise 3 groups. The first is proteinase, which initially cleaves the milk protein to peptides. The second group consists of transport systems for the internalization of oligopeptides, which are involved in the cellular uptake of small peptides and amino acids. The third group, peptidases in the cell, cleaves peptides into smaller peptides and amino acids. This review is to provide the information about the proteolytic system of LAB.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.221-227
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• 2006

BSCX1 was an antimicrobial peptide produced by Bacillus subtilis cx1. Attempts were made to determine the location of inducing factor in the bacteriocin-sensitive cell affecting bacteriocin BSCX1 production. Mixed culture of the bacteriocin producer strain B. subtilis cx1 and its sensitive strain B. subtilis ATCC6633, increased production of bacteriocin BSCX1. The result suggested the presence of a bacteriocin inducing factor in the sensitive strain. The inducing factor was localized in the cell debris and intracellular fraction of B. subtilis ATCC6633. Bacteriocin BSCX1 inducing factor was found to be highly stable in the pH range 2.5-9.5, but inactivated within 3h over $50^{\circ}C$, and treatment with proteinase K destroyed its inducing activity, this result suggested that the inducing factor should be a proteinaceous nature.

• Journal of Ginseng Research
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• v.13 no.1
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• pp.92-97
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• 1989

This study investigated the effects of high light intensity (100 KLw) and high temperature (45 ℃, dark) on enzyme (glucose-6-phosphate dehydrogenase, acid phosphatase, catalase, peroxidase, and proteinase) activities and characteristics of Panax ginseng C.A. Meyer leaves. Enzyme activity and protein content decreased rapidly under treatment with high light intensity In P ginseng the thermal stabilities of catalase and peroxidase were high (above 70%), and the coagulation rates of soluble proteins were low (below 17%). Therefore, the decrease in enzyme activity and protein content was not caused by increase in leaf temperature due to the high light intensity, but by increase in proteolytic activities. The photochemical formation rate of superoxide radical (O-2) was higher in the P ginseng leaf extracts than in Solanum nigmm, and was accelerated by addition of crude saponin to the buffer extracts.

• Journal of Plant Biology
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• v.38 no.4
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• pp.381-388
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• 1995

Four cONA clones expressed in late root nodules of Canavalia lineata were isolated by differential screening using total RNA from uninfected roots, Clb1 and uricase II cONAs as competitors and named Cnod1, Cne2, Cne3 and Clb2, respectively. Cnod1, hybridized to 1450 nt mRNA, was highly homologous to cysteine proteinase gene from rice and showed nodule-specific expression, especially in late nodules. Cne2, hybrdized to 900 nt mRNA, was moderately homologous to Expressed Sequence Tag of rice and expressed mainly in root nodules. Its expression was increased at 13 OAI and subsequently remained at the same level. Cne3, hybridized to 1700 nt and 1400 ot mRNAs, was highly homologous to tonoplast membrane intrinsic protein TRG31 gene from pea and was expressed strongly in roots and nodules, but weakly in leaves. Temporal expression pattern of Cne3 was coincided with the life cycle of root nodules. Clb2, hybridized to 800 nt mRNA, was expressed from 8 OAI, amplified at 13 DAI and remained steady thereafter.eafter.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.04a
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• pp.142-142
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• 2003

본 연구에서는 kiwifruit 과육 속에 들어 있는 단백질분해효소의 gelatin분해활성을 조사하고 그 산업적 방안을 검토하였다. Kiwifruit 과육에서 3개의 단백질분해효소의 활성 밴드(PI, PII, PIII)가 관찰되었다. 단백질분해효소 PI은 220 kD, PII는 51 kD, PIII는 26 kD에 해당하는 것으로 추정할 수 있었다. 이들 단백질분해효소 PI, PII, PIII는 모두 pH 2.0~5.0 범위에서 높은 활성을 보였으며 pH 4.0에서 가장 높게 나타났다. 이들 단백질분해효소 PI, PII, PIII는 모두 cysteine proteinase 저해제인 E-64와 iodoacetate에 의해서 저해되었으며, cysteine proteinase를 촉진하는 DTT, cysteine 및 $\beta$-mercaptoethanol에 의해서 활성이 증가하였다. 그 중 단백질분해효소 PIII는 분자량과 효소의 특성으로 보아 actinidin (EC 3.4.22.14)과 동일한 것으로 판단되었다. 단백질분해효소 PI, PII, PIII는 모두 $Ca^{2+}$, $Mg^{2+}$과 $Mn^{2+}$에 의해 촉진되었으며, $Zn^{2+}$과 Hg$^{2+}$에 의해 완전히 저해되는 것으로 나타났다. 하지만, Co$^{2+}$, Cu$^{2+}$, $Al^{3+}$ , Fe$^{3+}$ 등 금속이온의 영향은 다소 다르게 나타났다. Kiwifruit 과육의 단백질분해효소 PI, PII, PIII 중에서 PI과 PII는 온도가 증가함에 따라 활성이 점차 낮아졌으나 PIII는 비교적 안정한 것으로 조사되었다. 특히, PIII는 5$0^{\circ}C$ 이내의 범위에서 48시간 경과시에도 75% 이상의 활성을 보여 이 범위의 온도에서는 상당 시간 동안 안정한 것으로 나타났다. 단백질분해효소의 산업적 가치를 고려해 볼 때 우선적으로 넓은 기질특이성과 열안정성이 높아야 한다. Kiwifruit에서 추출한 단백질분해효소는 4$0^{\circ}C$ 전후에서 최대의 활성을 보이고, 고온에서도 상당 시간 비교적 안정한 특성을 보여 식품제조, 식육연화 등 식품산업 분야에서의 활용가능성이 높을 것으로 보이며, 나아가 단백질이 갖는 식품학적 기능성을 높이는 데에도 사용할 수 있을 것으로 판단된다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.4
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• pp.509-516
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• 2009

The CJ6 bacterial strain, which possesses strong antifungal activity, was isolated from meju and identified as Bacillus polyfermenticus based on Gram staining, biochemical properties, and 16S rRNA gene sequencing. B. polyfermenticus CJ6 showed antimicrobial activity against the various pathogenic molds, yeasts, and bacteria. Antifungal activity from B. polyfermenticus CJ6 was reduced after 24 hr at $70^{\circ}C$ but antifungal activity was not completely destroyed. The antifungal activity was stable in the pH range of $3.0{\sim}9.0$, and inactivated by proteinase K, protease, and ${\alpha}$-chymotrypsin, which indicate its proteinaceous nature. The apparent molecular masses of the partially purified antifungal compound, as indicated by using the direct detection method in Tricine-SDS-PAGE, was approximately 1.4 kDa.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.50-55
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• 2001

A new bacteriocin produced by Bacillus subtilis cx1, was partially purified and characterized. The bactericoin from B. subtilis cx1 was stable in the range of pH 2.5-9.5. B. subtilis csx1 retained its antimicrobial activity to long-term exposure at $-20^{\circ}C$ and $-70^{\circ}C$. However, B. subtilis cx1 was inactivated completely within 15 min over $60^{\circ}C$ and lost 50% of its antimicrobial activity within 15 min at $50^{\circ}C$, B. subtilis cx1 was inactivated by protease, trypsin, proteinase K and carboxypeptidase, which indi-cates its protein nature. Direct detection of the antimicrobial activity on Tricine -SDS-PAGE suggested an apparent molecular mass of about 9,500 dalton.

• Journal of Microbiology and Biotechnology
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• v.31 no.5
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• pp.705-709
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• 2021

Porphyromonas gingivalis (P. gingivalis) is a major bacterial pathogen that causes periodontitis, a chronic inflammatory disease of tissues around the teeth. Periodontitis is known to be related to other diseases, such as oral cancer, Alzheimer's disease, and rheumatism. Thus, a precise and sensitive test to detect P. gingivalis is necessary for the early diagnosis of periodontitis. The objective of this study was to optimize a rapid visual detection system for P. gingivalis. First, we performed a visual membrane immunoassay using 3,3',5,5'-tetramethylbenzidine (TMB; blue) and coating and detection antibodies that could bind to the host laboratory strain, ATCC 33277. Antibodies against the P. gingivalis surface adhesion molecules RgpB (arginine proteinase) and Kgp (lysine proteinase) were determined to be the most specific coating and detection antibodies, respectively. Using these two selected antibodies, the streptavidin-horseradish peroxidase (HRP) reaction was performed using a nitrocellulose membrane and visualized with a detection range of 10³-10⁵ bacterial cells/ml following incubation for 15 min. These selected conditions were applied to test other oral bacteria, and the results showed that P. gingivalis could be detected without cross-reactivity to other bacteria, including Streptococcus mutans and Escherichia fergusonii. Furthermore, three clinical strains of P. gingivalis, KCOM 2880, KCOM 2803, and KCOM 3190, were also recognized using this optimized enzyme immunoassay (EIA) system. To conclude, we established optimized conditions for P. gingivalis detection with specificity, accuracy, and sensitivity. These results could be utilized to manufacture economical and rapid detection kits for P. gingivalis.