• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.181-186
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• 2000

MAPI (microbial alkaline protease inhibitor) was isolated from cultrue broth of Streptomyces chromofuscus SMF28. The Ki values of MAPI for the representative serine proteases such as chymotrypsin and proteinase K were 0.28 and $0.63{\;}\mu\textrm{M}$, respectively, and for the cysteine proteases cathepsin B and papain were 0.66 and $0.28{\;}\mu\textrm{M}$, respectively. These data indicate that MAPI is not a potent selective inhibitor of serine or cysteine proteases. Progress curves for the inhibition of three proteases by MAPI exhibithe characteristic patterns; MAPI exhibited slow-binding inhibition of cathepsin B. It was rapidly associated with chymotrypsin before the addition of substrate and then reactivation of MAPI-inhibited enzyme was investigated in the presence of substrate. On the other hand, MAPI-proteinase K interaction was typical for those classical inhibitors. When MAPI was incubated with trypsin, there was an extensive reduction in the ingibitory activities of MAPI corresponding to 66.5% inactivation of MAPI, indicating that trypsin-like protease may play a role in the decrease of the inhibitory activity during cultivation.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.203-210
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• 1997

The difrrrenlial display reverse transcription polymerase chain reaction (DDRT-PCR) aniilysis roils performed to identify the pathogellir strain specific amplicons. mRNAs were purified from the trophozoites of the pathogenif strain YS-27 and the non-pathogenic strain S 16. respectively. Three kinds of rirsl stranded rDNAs were reverse transcribed from the mRNAs by one base anchored oligo-dT 11M (M: A. C, or G) primers. Each cDNA lemplatr was used for DDRT-PCK analysis. A total of 144 pathogenic strain specific amplicons was observed in DDRT-PCR analysis using primer combinations of the 11 arbitrary primers and the 3 one base anchored oli해-dT11M primers. Of these 31 amplit'tons were verified as the amplirons amplified only from the mRNAs of the pathogenic strain by DNA slots biol llybridizatioil. Furthel cklaracleization of the 31 pathogenic strain sprcifil amplicons by DNA slot blot hybridlnation analysis using biotin labeled Probes or the PCR amplified DNA of rysteine proteinase genes revealed that 21 of them were amplliried from the maNAs of the cysteine proteinase genes. Four randomly selected amplirons out of the rest 10 amplirons were used fur screening of cDNA library followed by immunoscreening and all of them were turned outs to be amplified from the mRNA.

• Journal of Plant Biotechnology
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• v.2 no.2
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• pp.83-87
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• 2000

Carnation petals exhibit autocatalytic ethylene production and wilting during senescence. The autocatalytic ethylene production is induced by the expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase genes, whereas the wilting of petals is related to expression of the cysteine proteinase (CP) gene. Until recently, it has been believed that these two phenomena, autocatalytic ethylene production and wilting, are regulated in concert in senescing carnation petals, since the two phenomena occurred closely in parallel. Our studies with petals of a transgenic carnation harboring a sense ACC oxidase transgene and petals of carnation flowers treated with 1,1-dimethyl-4-(phenylsulfonyl) semicarbazide showed that the expression of ACC synthase and ACC oxidase genes and that of CP are regulated differently in carnation psanetals. Interestingly, in the petals of transgenic carnation, the transcript for CP was accumulated but the transcripts for ACC synthase and ACC oxidase were not accumulated in response to exogenous ethylene. Based on these results, we hypothesized that two ethylene signaling pathways, one leading to the expression of ACC synthase and ACC oxidase genes and the other leading to the expression of CP gene, are functioning in senescing carnation petals.

• 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.

• Korean Journal of Plant Resources
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• v.11 no.2
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• pp.195-201
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• 1998

Some morphological character were surveyed and PR-proteinases were induced and characterized from Lilium formosanum Wallace endeimc to Cheju island . Its flower characters were similar to those of white trumpet lilies(Lilium longiflorum Thunb) although its flowering period was later than that of white trumpet lilies and it hadd a wide range of variation among individuals. Six PR-proteinases(II-2, III-1, III-2, IV-1, IV-2 and V) were induced from bulbs by 2.5mM salicylic acid and almost excreted into the intercellular spaces. These PR-proteinases were strongly activated by Ca 2+, , whereas they were strongly inhibited by Cu2+ Co2+ and Fe2+ . Three PR proteinases(II-2, IV-1 and IV-2) were strongly inhibited by 1, 10 -phenanthroline, indicating that these enzymes are metallo-proteinases. Three PR-proteinases(III-1, III-2 and V) had a high sensitivity to PMSF and required $\beta$-mercaptoethanol for their activities. These results indicate that these proteinases are cysteine proteinases.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.11a
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• pp.232-232
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• 2005

• Proceedings of the PSK Conference
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• 2000.10a
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• pp.195.2-196
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• 2000

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.05a
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• pp.148-149
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• 2001

Protein inhibitors are proteins or peptides capable of inhibiting catalytic activities of proteolytic enzymes. They are grouped primarily as either serine, cysteine, aspartic or metallto-proteinase inhibitors. Pretense inhibitors have been hewn since the end of the last century in nematodes and human blood serum, and their ubiquitous distribution in microorganisms, animals and plants has been widely documented. (omitted)

• Proceedings of the Korean Society of Crop Science Conference
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• 2023.04a
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• pp.133-133
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• 2023

• 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.