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( Won Tae Kim ),( Sung Woo Bae ),( A Young Kim ),( Kwan Ho Park ),( Sang Beom Lee ),( Young Cheol Choi ),( Sang Mi Han ),( Young Han Park ),( Young Ho Koh ) 생화학분자생물학회(구 한국생화학분자생물학회) 2011 BMB Reports Vol.44 No.6
To investigate the molecular scavenging capabilities of the larvae of Hermetia illucens, two serine proteases (SPs) were cloned and characterized. Multiple sequence alignments and phylogenetic tree analysis of the deduced amino acid sequences of Hi-SP1 and Hi-SP2 were suggested that Hi-SP1 may be a chymotrypsin-and Hi-SP2 may be a trypsin-like protease. Hi-SP1 and Hi-SP2 3-D homology models revealed that a catalytic triad, three disulfide bonds, and a substrate-binding pocket were highly conserved, as would be expected of a SP. E. coli expressed Hi-SP1 and Hi-SP2 showed chymotrypsin or trypsin activities, respectively. Hi-SP2 mRNAs were consistently expressed during larval development. In contrast, the expression of Hi-SP1 mRNA fluctuated between feeding and molting stages and disappeared at the pupal stages. These expression pattern differences suggest that Hi-SP1 may be a larval specific chymotrypsin-like protease involved with food digestion, while Hi-SP2 may be a trypsin-like protease with diverse functions at different stages. [BMB reports 2011; 44(6): 387-392]
Kim, In-Seop,Kang, Sung-Gyun,Lee, Kye-Joon The Microbiological Society of Korea 1995 The journal of microbiology Vol.33 No.4
The relationship between morphological differentiation and production of trypsin-like protease (TLP_ in streptomycetes was studied. All the Streptomyces spp.In this study produced TLP just before the onset of aerial mycelium formation. Addition of TLP inhibitor, TLCK, to the top surface of colonies inhibited aerial mycelium formation as well as TLP inhibitor, TLCK, to the top surface of colonies inhibited aerial mycelium formation as well as TLP activity. Addition of 2% glucose to the Bennett agar medium repressed both the aerial mycelium formation and TLP production in S. abuvaviensis, S. coelicolor A3(2), S exfoliatus, S. microflavus, S. roseus, s. lavendulae, and S. rochei. However the addition of glucose did not affect S. limosus, S. felleus, S. griseus, S. phaechromogenes, and S. rimosus. The glucose repression on aerial mycelium formation and production of TLP was relieved by the addition of glucose anti-metabolite (methyl .alpha.-glucopyranoside). Therefore, it was concluded that TLP production is coordinately regulated with morphological differentiation and TLP activity is essential for morphological differentiation in streptomycetes. The proposed role of TLP is that TLP participates in the degradation of substrate mycelium protein for providing nutrient for aerial mycelial growth.
Kim, In Seop,Lee, Kye Joon 한국미생물 · 생명공학회 1995 Journal of microbiology and biotechnology Vol.5 No.4
Nutritional factors regulating the morphological differentiation and physiological differentiation of Streptomyces exfoliatus SMF13 on surface cultures were evaluated. S. exfoliatus SMF13 produced leupeptin and chymotrypsin-like protease (CTP) at the stage of substrate mycelium growth, and leupeptin-inactivating enzyme (LIE) and trypsin-like protease (TLP) at the stage of aerial mycelium growth. The activity of leupeptin and CTP was high in the region of active growing substrate mycelium, whereas the activity of LIE and TLP was high in the region of aerial mycelium or spores. The differentiations were induced in glucose-limited conditions or by the addition of glucose anti-metabolite (methyl α-glucopyranoside), but repressed by high concentrations of glucose or casamino acids. Morphological differentiation (formation of aerial mycelia and spores) was closely related with physiological differentiation (formation of brown-pigment, LIE and TLP). The local distribution of leupeptin, CTP, LIE, and TLP in a developing colony showed that colony development correlated with the production and functions of the compounds: CTP is essential for providing a nitrogen source for mycelium growth: leupeptin regulates TLP activity: LIE inactivates leupeptin: TLP hydrolyzes nongrowing mycelium.
Inhibition of Various Proteases by MAPI and Inactivation of MAPI by Trypsin
LEE, HYUN SOOK,KHO, YUNG HEE,LEE, KYE JOON 한국미생물 · 생명공학회 2000 Journal of microbiology and biotechnology Vol.10 No.2
MAPI (microbial alkaline protease inhibitor) was isolated from culture broth of Streptomyces chromofuscus SMF28. The K_i values of MAPI for the representative serine proteases such as chymotrypsin and proteinase K were 0.28 and 0.63μM, respectively, and for the cysteine proteases cathepsin B and papain were 0.66 and 0.28μM, 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 exhibited 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 inhibitory 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.
이배익 ( Bae Ik Lee,),김이청 ( Yi Cheong Kim ),김근업 ( Geun Up Kim ),권오남 ( O Nam Kwon ) 한국수산과학회(구 한국수산학회) 2012 한국수산과학회지 Vol.45 No.1
We compared the nutritional requirements of whole larvae of the black seabream Acanthopagrus schlegeli, sweet fish Plecoglossus altivelis, olive flounder Paralichthys olivaeus and rock fish Sebastes schlegeli. The larvae were 20, 30, 14 and 5 DAH (or spawning) of black seabream, sweet fish, olive flounder and rock fish, respectively. Specific α-amylase activity (mU/mg protein) was highest (8,324.9 mU/mg protein) in rock fish larvae (P<0.05). Specific trypsin-like protease activity was highest (11,330.1 mU/mg protein) in black seabream larvae (P<0.05), which also exhibited the highest activity, 685.5 mU/mg dry weight (P<0.05). The specific activities per mg protein and mg dry weight of black seabream were the highest (187.4 mU/mg protein and 11.3 mU/mg dry weight, respectively) (P<0.05). A/P, P/L and A/L ratios of rock fish were 1.47, 90.3 and 133.1, respectively (P<0.05). We present here basic larval digestive enzymatic nutritional requirement data.