• BMB Reports
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• v.32 no.6
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• pp.579-584
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• 1999

Mantis egg fibrolase (MEF-3) was purified from the egg cases of Tenodera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, DEAE Affi-Gel blue gel affinity chromatogragphy, and MONO-Q anion-exchange chromatography. This protease had a molecular weight of 35,600 Da as determined by SDS-polyacrylamide gel electrophoresis under reducing conditions and its isoelectric point was 6.0. The N-terminal amino acids sequence was Ala-Thr-Gln-Asp-Asp-Ala-Pro-Pro-Gly-Leu-Ala-Arg-Arg. This sequence was 80% homologous to the serine protease from Tritirachium album. MEF-3 readily digested the ${\alpha}$-and ${\beta}$-chains of fibrinogen and more slowly the ${\gamma}$-chains. It showed strong proteolytic and fibrinolytic activities. Phenylmethanesulfonyl fluoride and chymostatin inhibited its proteolytic activity, while EDTA, EGTA, cysteine, ${\beta}$-mercaptoethanol, elastinal, tosyl-lysine chloromethylketone, and tosyl-amido-2-phenylethyl chloromethyl ketone did not affect its proteolytic activity. Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEF-3 was benzoyl-Phe-Val-Arg-p-nitroanilide. Based on these experimental results, we speculated that MEF-3 is a serine protease with a strong fibrin(ogen)olytic activity.

• Ocean and Polar Research
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• v.30 no.4
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• pp.467-472
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• 2008

Antarctic krill has a strong proteolytic enzyme system, which comes from a combination of several proteases. This powerful activity can be easily detected by krill's superior post mortem autolysis. Mammalian skin consists of epidermis and dermal connective tissue, and functions as a barrier against threatening environments. A clot in a wound site of the skin should be removed for successful skin regeneration. Epithelial cells secrete proteases to dissolve the clot. In previous studies Antarctic krill proteases were purified and characterized. The proteolytic enzymes from Antarctic krill showed higher activity than mammalian enzymes. It has been suggested that these krill clean up the necrotic skin wound to induce a natural healing ability. The enzymes exhibited additional possibilities for several other biomedical applications, including dental plaque controlling agent and healing agent for corneal alkali burn. Considering that these versatile activities come from a mixture of several enzymes, discovering other proteolytic enzymes could be another feasible way to enhance the activity if they can be used together with krill enzymes. Molecular cloning of the krill proteases should be carried out to study and develop the applications. This review introduces possible roles of the unique Antarctic krill proteases, with basic information and suggestion for the development of an application to skin regeneration.

• Microbiology and Biotechnology Letters
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• v.14 no.6
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• pp.467-471
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• 1986

Yeast cell wall lytic enzyme was purified from Fusarium moniliforme by ammonium sulfate fractionation and gel column chromatography. The lytic activity was found to consist of three enzyme activities which were resolved on Sephadex G-100. The first peak on chromatogram exhibited proteolytic, lytic and laminarinase activities, and the second had both lytic and laminarinase activities, whereas the third peak was shown to contain lytic activity only. Three enzyme activities showed the synergistic effect and reducing agents accelerated the yeast roil wall lysis. This indicates that lytic, proteolytic and laminarinase activity acted cooperatively in the lysis of intact cells. Tannic acid precipitate of crude enzyme constituted of three enzyme activities had a high lytic activity on viable yeast cell and has proved useful in yeast protoplast formation.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.167-170
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• 2000

Proteolytic hydrolysis is one of the main enzymatic reaction of waste activated sludge (WAS) digestion. Pretense excreted from Bacillus stearothermophilus (ATCC 31197) showed optimum temperature of $75^{\circ}C$ for maxium heat stable proteolytic activity against azo casein. The dependency of $Ca^{2+}$, $Zn^{2+}$ on heat stability of proteolytic enzymes were measured with various concentrations. It was shown that $Ca^{2+}$ ion enhanced heat stability of these enzymes. Then thermophilic aerobic digestion (TAD) was performed using B. sterothermophilus with the addition of divalent ions. Performance of TAD process with ATCC 31197 activated by $Ca^{2+}$, $Zn^{2+}$ions in terms of dissolved organic carbon (DOC) concentration, extracellular protein concentration, and scanning electrion microscopy (SEM) analysis. The best result of protein reduction concentration in digestion test was obtained with the addition of 2 mM $Ca^{2+}$ ion.

• Food Science of Animal Resources
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• v.30 no.4
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• pp.669-673
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• 2010

The purpose of this study was to evaluate kiwifruit as a tenderizer by its effects in improving the quality of various beef parts. Basic data are provided for development of standard recipes for convenient cooking in foodservice. The optimum temperature of the crude proteolytic enzymes in the kiwifruit was determined to be $65^{\circ}C$. The substrate specificity of the enzymes was higher in beef than in pork, chicken, or duck. The enzymes had their greatest effects on chuck and rib parts, and had lower effects on loin, breast, and round parts, in that order. As the amount of kiwifruit extract increased, the moisture content of the cooked beef also increased. The addition of 10% kiwifruit improved the sensory quality of the cooked beef. In particular, the texture scores of cooked beef samples treated with 10% and 15% kiwifruit extract were significantly higher than the other samples (p<0.001), and juiciness and overall acceptability scores were also highest. In summary, 10% kiwifruit extract is deemed an appropriate addition to improve.

• BMB Reports
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• v.36 no.4
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• pp.390-398
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• 2003

The CDP/Cux transcription factor was previously shown to be proteolytically processed at the G1/S transition. In view of characterizing and eventually identifying the protease responsible for CDP/Cux processing, we have established an in vitro proteolytic processing assay. CDP/Cux recombinant proteins expressed in mammalian or bacterial cells were efficiently processed in vitro using as a source of protease either whole cell extracts, the nuclear or the cytoplasmic fraction. Processing was found to take place optimally at a lower pH, to be insensitive to variations in salt concentration, and to be inhibited by the protease inhibitors MG132 and E64D. Interestingly, the bacterially-produced substrate was more efficiently processed than the substrate purified from mammalian cells. Moreover, processing in vitro was more efficient when CDP/Cux substrates were purified from populations of cells enriched in the S phase than in the G1 phase of the cell cycle. Altogether, these results suggest that post-translational modifications of CDP/Cux in mammalian cells inhibits processing and contributes to the cell cycle-dependent regulation of processing. The in vitro processing assay described in this study will provide a useful tool for the purification and identification of the protease responsible for the processing of CDP/Cux.

• Fisheries and Aquatic Sciences
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• v.18 no.4
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• pp.349-357
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• 2015

To reutilize fisheries waste, we isolated a bacterial strain from a coastal area located in Busan. It was identified as Bacillus licheniformis TK3-Y. Using plate assay and 500-mL flask experiments, we found that the isolate simultaneously possessed cellulolytic, proteolytic, and lipolytic activities with salt tolerance. 10% (v/v) inoculums, were used to examine the biodegradation characteristics of the TK3-Y strain on carboxymethylcellulose, skim milk, and olive oil media. The optimum conditions for pH, temperature, agitation speed, and NaCl concentration on each 1% substrate were 6, $50^{\circ}C$, 180 rpm, and 17.5%, respectively. Under optimal conditions, the TK3-Y strain showed 1.07 U/mL cellulolytic, 1,426 U/mL proteolytic, and 6.45 U/mL lipolytic activities. Each enzyme was stable within a range of 17.5-35% NaCl. Therefore, the salt tolerance ability of strain TK3-Y was superior to other related strains. In degradation of a mixed medium containing all three substrates, both the cellulolytic and proteolytic activities were somewhat lower than those on each single substrate, while the lipolytic activity was somewhat higher. From the above results, the TK3-Y strain appears to be a good candidate for use in the efficient treatment of fisheries waste in which components are not collected separately.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.6
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• pp.803-806
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• 1993

The properties of proteolytic enzymes from the fruit of Broussonetia Kazinoki Siebold were investigated. The protease activity of the enzymes from the fruit of Broussonetia Kazinoki Siebold was 1.6 unit. The optimum temperature and pH of the enzymes were $60^{\circ}C$ and 7.0, respectively. The enzymes were stable at pH values from 6 to 8 for 1 hr. at $37^{\circ}C$ of incubation and also retained all activity after incubation for 1 hr. at $60^{\circ}C$. The enzyme preparations showed strong activities toward hemoglobin and collagen.

• Journal of Pharmaceutical Investigation
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• v.19 no.1
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• pp.9-14
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• 1989

In order to study the protease from Agaricus bisporus (Lange), the crude protease preparation was separated by fractionation of mushroom extracts with ammonium sulfate. It was found that extracts from Agaricus bisporus (Lange) Sing. contained protease. The optimum pH of the enzyme was 6.0, and the pH range at which the enzyme was stable was 4.0 to 7.0. The optimum temperature at which the enzyme showed the highest proteolytic activity was $50^{\circ}C$, while the enzyme was instantly inactivated at about $60^{\circ}C$. The enzyme activity was inhibited by $Ag^+$, $Hg^{2+}$, $Cu^{2+}$, $Ba^{2+}$, $Fe^{3+}$, $Co^{2+}$, $Ca^{2+}$, $Pb^{2+}$, $Mg^{2+}$ and $Mn^{2+}$. The $K_m$ value was 0.32 mM with Hammarsten casein.

• Food Science and Biotechnology
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• v.14 no.5
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• pp.614-620
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• 2005

The antioxidative effect of Ecklonia cava, a brown marine alga, was investigated on radical scavenging, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), and hydroxyl and alkyl radicals, using an electron spin resonance (ESR) technique, and on the inhibition of $H_2O_2$-induced DNA damage using comet assay. E. cava was enzymatically hydrolyzed with five food industrial proteases (Alcalase, Flavourzyme, Kojizyme, Neutrase and Protamex) to prepare water-soluble extracts. All the proteolytic hydrolysates exhibited strong dose-dependent radical scavenging activities (above 80%) at a concentration of $2.5\;{\mu}g/mL$. Kojizyme extract (obtained by proteolytic hydrolysation of E. cava with Kojizyme) showed the highest hydroxyl radical scavenging activity of around 98%. In addition, the $H_2O_2$-induced DNA damage was determined using a comet assay, which was quantified by measuring the tail length. Reduction of DNA damage increased with increasing concentrations of Kojizyme extract from E. cava. These results indicated that E. cava has a potential as a valuable natural antioxidative source.