• Parasites, Hosts and Diseases
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• v.34 no.1
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• pp.49-58
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• 1996

Characterization of a purified proteinase from T4chomoncs uoginalis was carried out using bacitracin-sepharose affinity chromatography. Trichomonos uqginolis KT-9 isolate was used as a source of eye study Proteinase activity was determined using Bz-Pro- Phe-Arg-Nan as the substrate. Optimum pH for the purified proteinase activity was 7.0 and 6.0, 9.0 with DTT. Optimum temperature was 37℃ and isoelectric point was 7.2 Activity of this proteinase was inhibited by E-64, antipain, leupeptin, Hg2+ and Zn2+ and activated by DTT and cysteine. Activity of the purified proteinase was visualized by gelatin SDS- PAGE. The gelatinolytic activity of the purified proteinase was inhibited by E-64, antipain, leupeptin, and IAA, but not by PMSF and EDTA. On SDS-PAGE, the molecular weight of the purified proteinase was 60,000 daltons. Sera of rabbits infected with T. vaginalis reacted specifically in immunoblots with this proteinase. These results indicate that 60 kDa of purified proteinase was cysteine proteinase with antigenicity.

• Journal of Nutrition and Health
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• v.6 no.4
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• pp.55-60
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• 1973

To determine the effects of condiments upon Proteinase Activity, condiments such as welsh onion, garlic, ginger, black pepper, red pepper, mi-won (glutamic acid natrium), sugar, mustard and horse-radish were ground by a homogenizer, and each of them was dosed by 0%, 1%, 5% and 10% into Pancreatin Solution of 0.2% for storage at the temperature of 15 degrees Cels. The Enzyme Solution thus obtained then was measured at a certain interval of time by the Fuld Gross Method, and the following results were obtained. 1) The condiments that kept Proteinase Action of Pancreatin checked below 75% were mustard, horse-radish, red pepper and welsh onion. The centrol power of welsh onion, in particular, became stronger as storage time became longer. 2) The condiments that kept Proteinase Action of Pancreatin checked below 50% were sugar, black pepper and ginger. 3) Mi-won and garlic showed a strong checking powor over Proteinase Action at an early stage of storage, but as time passed, their control power gradually diminished to naught. In short, it may be concluded that ail of the condiments used in this experiment demonstrated their checking power over Proteinase Action.

• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.381-385
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• 1996

Three distinct plasmids, with approximate molecular weights of 1, 4.5, and 33 megadaltons, were found in Lactococcus lactis subsp. lactis (L. lactis) ML8. Slow acid-producing mutants of L. lactis ML8, isolated by plasmid curing with acriflavine treatment, lacked the 33-megadalton plasmids. The plasmid-cured mutant showed lactose-negative (Lac) characteristics and the alteration of extracellular proteinase pattern. The possible involvement of extracellular proteinase with the 33-megadalton plasmid is highlighted in this research.

• Food Science of Animal Resources
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• v.30 no.3
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• pp.443-448
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• 2010

Casein phosphopeptide (CPP) enhances calcium absorption in humans. Lactic acid bacteria (LAB) are capable of synthesis of cell-surface proteinase, which can hydrolyze milk protein and release several types of peptides in the medium. This study was conducted to characterize proteinase of LAB and to evaluate the CPP production from bovine milk. The content of CPP of milk produced by cell-free extract of LAB was determined based on the quantity of decomposed peptide from casein using the O-phthaldialdehyde (OPA) method. The proteolytic activity of LAB was assayed using fluorescein isothiocyanate (FITC)-labeled casein. Casein appeared to be a better substrate than whey proteins for extracellular proteinases of LAB. During fermentation, milk proteins were hydrolyzed by extracellular proteinase of LAB, resulting in an increase in the amount of free $NH_3$ groups. Overall, the results presented here indicate that CPP produced by LAB may be a promising material for novel applications in the dairy industry.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.75-81
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• 1998

The yeast strain 504D, isolated from salted shrimp soup, showed the best proteolytic activity under alkaline condition. The yeast formed vegetative cells in almost optimal media for yeasts, but formed only pseudohyphae in the MM medium containing citric acid and true hyphae in the MM medium containing N-acetylglucosamin and ${\beta}$-D-glucose. The yeast was classified as hemiascomycetes to form ascospores by sexual reproduction, and formed blastospores and athrospores by asexual reproduction. The yeast strain did not assimilate almost of the carbon sources, nitrate and nitrite, but some organic acids and alcohols. The fatty acids of whole cells were composed of 53.67% unsaturated fatty acids and 14.58% saturated, and, especially, C17:1 was observed in this strain but not in two control yeasts. However, almost of all results were very similar to the morphological and physiological characteristics of Yarrowia lipolytica KCCM 12495 and KCCM 35426, except for a little differences which are the composition of fatty acids and the manner of mycellium formation. Therefore, the isolated yeast strain 504D is identified as a Yarrowia lipolytica.

• BMB Reports
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• v.28 no.1
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• pp.68-71
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• 1995

Bovine gastricsin catalyzes peptide synthesis over an optimum range of pH 4~5, resulting in satisfactory yields of methyl esters and p-nitroanilides of benzyloxycarbonyl tetra- to hexa-peptides, provided that hydrophobic amino acid residues form new peptide bonds. The effectiveness of the enzyme also depends on the nature of adjacent amino acid residues. An aspartic proteinase with a characteristic gastricsin specificity pattern would be useful for the synthesis of middle-length peptides.

• BMB Reports
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• v.30 no.6
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• pp.448-452
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• 1997

Polyclonal antibody of the boar sperminogen was used to characterize the boar sperm proteinase sperminogen. Boar sperminogen was purified from the acid extracts of the washed epididymal spermatozoa by gel filtration through a Sephadex G-100 column. followed by preparative SDS-PAGE. The sperminogen band was sliced out and was eluted from the gel matrix. The purified sperminogen was used to produce the polyclonal antibody of the boar sperminogen. When characterized on a Western blot, the final preparation of sperminogen appeared as a homogenous protein with a molecular weight of 32 kDa. The relative migration of sperminogen was distinctly different from the major components of the proacrosin-acrosin system as well as all the observable proacrosin activation by-products detected on the Western blot. The sperminogen antibody, however. cross-reacted with the proacrosin-acrosin system.

• Animal cells and systems
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• v.3 no.2
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• pp.221-228
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• 1999

In order to find a biochemical marker for late Iysosomes, we characterized two cDNAs which were cloned by using a monoclonal antibody (mAb) against Iysosomes in Amoeba proteus as a probe. The two cDNAs, a 1.3-kb cDNA in pBSK-Iys45 and a 1.6-kb cDNA in pBSK-Iys60, were found to encode proteins homologous to pepstatin-insensitive carboxyl proteinases (PICPs). E. coli transformed with pBSK-Iys45 produced two immunopositive polypeptides (45 and 43 kDa) and the cDNA in 1274 bases encoded a 44,733-Da protein (Lys45) of 420 amino acids containing one site for a core oligosaccharide. On the other hand, E. coli transformed with pBSK-Iys60 produced several polypeptides (64, 54, 45, 41, and 37 kDa) reacting with the mAb. The cDNA contained 1629 bases and encoded a 59,231-Da protein (Lys60) of 530 amino acids containing two sites for asparagine-linked core oligosaccharides. These two cDNAs showed identities of 60.3% in nucleotide sequences and 23.6% in amino acid sequences. Lys45 and Lys60 appeared to share XXEFQK as a common antigenic domain. The amino acid sequence of the Lys45 protein showed 17.4% identity and 40.9% similarity to that of PICP from Pseudomonas sp. 101. On the other hand, Lys60 showed a 24.3% identity and 51.9% similarity with human Iysosomal PICP in the amino acid sequence. A putative active center for serine protease, GTS＊xxxxxFxG, was found to be conserved among PICP homologues. The two PICPs are the first reported enzymatic markers for late Iysosomes.

• Korean Journal of Food Science and Technology
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• v.7 no.4
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• pp.229-237
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• 1975

An attempt was made to utilize the enzyme produced by Asp. oryzae as meat tenderizer. The production, purification, and various properties of proteinase produced by Asp. oryzae were investigated. Results obtained are as follow; 1. A strain which had the highest proteolytic activity was selected among 9 Aspergillus species. 2. Culture medium consisted of wheat bran 10g, 2% glucose, 0.03% urea and 0.1% $MgSO_4$ (pH 6.5). Mold was incubated at $30^{\circ}C$ for 3 days. 3. Enzyme extract from culture medium were fractionated with ammonium sulfate and purified by Sephadex G-75 column chromatography. 4. When pH of reaction mixture was controlled, maximal activity of proteinase by Asp. oryzae was obtained at pH 3, pH 6.6, $8.4{\sim}8.5$ and pH 10.0 to 10.5. Those results were interpreted to show that enzyme consists of acid proteinase, neutral proteinase and alkaline proteinase. Enzyme was stable at pH 6 to 10. 5. Opt. temperature for proteinase activity was $50^{\circ}C$, but enzyme was stable up to $40^{\circ}C$. 6. The proteinase was inhibited by $Ag^+$. It was also inhibited by EDTA. 7. When myofibrillar proteins were treated by proteinase from Asp. oryzae, ATPase activities of myofibrillar proteins changed remarkably. Accordingly, it was concluded that proteinase produced by Asp. oryzae were able to be used as meat tenderizer.

• Journal of Ginseng Research
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• v.32 no.4
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• pp.300-304
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• 2008

Cysteine proteinases play an essential role in plant growth and development but also in senescence and programmed cell death. They participate in both anabolic and catabolic processes. In addition, they are involved in signalling pathways and in the response to biotic and abiotic stresses. A cDNA clone encoding cysteine proteinase (CP) gene, designated PgCysP1, was isolated from Panax ginseng C. A. Meyer. Reverse transcriptase (RT)-PCR results showed that PgCysP1 expressed at different level in P. ginseng hairy root. Different stresses such as biotic as well as abiotic stresses triggered a significant induction of PgCysP1. The positive responses of PgCysP1 to the various stimuli suggested that PgCysP1 may help to protect the plant against reactive environmental stresses.