• Journal of Applied Biological Chemistry
• /
• v.62 no.4
• /
• pp.385-389
• /
• 2019

In this study, proteolytic enzymatic treatment conditions for Korean red ginseng were examined to increase the extraction yield. Commercially available proteases were screened to obtain high protein and carbohydrate yield. The optimal dosage and reaction time for Alcalase, the chosen protease, were found to be 2.0% (w/w) and 1.5 h, respectively. Treatment with optimal conditions of Alcalase increased solid yield, total phenolic content and gensenosides content by 57.6, 81.8, and 33.8%, respectively, over levels in non-treated Korean red ginseng. Antioxidative activities evaluated by free radical scavenging activity, cation radical scavenging activity and reducing power were exactly similar between Alcalase-treated and non-treated extracts.

• Korean Journal of Food Science and Technology
• /
• v.22 no.3
• /
• pp.234-240
• /
• 1990

Attempts have been made to optimize the hydrolysis conditions of the oyster and the mussel by the commercial proteolytic enzymes. Raw materials were digested with seven different commercial enzymes, and their quality parameters measured in terms of degree of hydrolysis and content of free amino nitrogen, nucleic acid-related substances. and free amino acids as well as sensory evaluation of optimization of their hydrolysis conditions. As a result, following enzymes have been disclosed as effective for enzymatic digestion: MKC-HT proteolytic, alcalase 0.6L and thermease for the oyster whereas MKC-acid fungal protease and thermoase for the mussel, respectively.

• The Korean Journal of Mycology
• /
• v.14 no.1
• /
• pp.25-30
• /
• 1986

The purposes of this study were to investigate enzyme components and its physiological activities of Sarcodon aspratus (Berk.) S. Ito which grows wildly in Korea, belonging to the family Thelephoraceae. The carpophores of the fungus was extracted with cooling distilled water and salted out by ammonium sulfate. The precipitate was purified by dialysing through visking tube against distilled water and then dissolved with pH 7.8 ammonia aqua, and the extract was filterated. The fraction of filtrate was obtained as light brown powder after lyophilization and determined proteolytic activity. Protease activity of Sarcodon aspratus (Berk.) S. Ito was about two-third of that of pepsin on casein by cup method. The proteolytic potency of this enzyme was found to be 500 unit/mg. This proved the efficacy of the mushroom when it was used as a folk medicine for treating indigestion of beef.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.5
• /
• pp.990-996
• /
• 2008

In a previous study, we found an antifungal effect on human pathogenic fungi by the cell-penetrating peptide Tat (47-58) derived from HIV-1. Tat (47-58) immediately entered into the fungal nucleus and affected some physiological changes on the intracellular condition. In this study, Tat (47-58) showed a broad spectrum of antibacterial activity against pathogenic bacteria including bacterial clinical isolates. To improve resistance against proteases for use in vivo, we synthesized an analog of Tat (47-58) by substituting the L-amino acid for the D-amino acid. The D-enantiomer of Tat (47-58) also exhibited a broad spectrum of antibacterial activity at almost the same level of L-Tat (47-58) concentration. Unlike L-Tat (47-58), D-Tat (47-58) showed a significant proteolytic resistance against all proteases tested and antimicrobial activities in the presence of trypsin. Moreover, D-Tat (47-58) inhibited MRSA infection in human HeLa cells whereas L-Tat (47-58) partially allowed MRSA infection, and the results were due to the proteolytic resistance of D-Tat (47-58).

• Microbiology and Biotechnology Letters
• /
• v.26 no.6
• /
• pp.490-496
• /
• 1998

Aspergillus oryzae producing high proteolytic enzyme was isolated from soybean koji and named tentatively A. oryzae O-1. A. oryzae U-1 was obtained by mutation of A. oryzae O-1 with ultraviolet (UV)-irradiation and produced 14 times higher pretense activity compared with A. oryzae O-1. A. oryzae E-1 was acquired by treatment of A. oryzae U-1 with 0.5 M ethylmethanesulfonate (EMS) for 6 min at 3$0^{\circ}C$ and produced 39 times higher proteolytic activity than A. oryzae O-1. With protoplast fusion between A. oryzae O-1 and A. oryzee E-1 in the presence of polyethylenegylcol (PEG)-CaCl$_2$, proteolytic activity was increased to 82 times compared to A. oryzae O-1, and the fusant was named A. oryzae PF. The activities of the cultures containing proteolytic enzymes produced by the strains were determined to be 0.23 U/$m\ell$ for A. oryzae O-1, 3.29 U/$m\ell$ for A. oryzae U-1, 8.91 U/$m\ell$ for A. oryzae E-1, and 19.0 U/$m\ell$ for A. oryzae PF.

• The Korean Journal of Physiology and Pharmacology
• /
• v.21 no.1
• /
• pp.19-26
• /
• 2017

We investigated whether betulin affects the gene expression, secretion and proteolytic activity of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as in vivo production of MMP-3 in the rat knee joint to evaluate the potential chondroprotective effect of betulin. Rabbit articular chondrocytes were cultured and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-$1{\beta}$ ($IL-1{\beta}$)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. Effect of betulin on IL-$1{\beta}$-induced secretion and proteolytic activity of MMP-3 was investigated using western blot analysis and casein zymography, respectively. Effect of betulin on MMP-3 protein production was also examined in vivo. The results were as follows: (1) betulin inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) betulin inhibited the secretion and proteolytic activity of MMP-3; (3) betulin suppressed the production of MMP-3 protein in vivo. These results suggest that betulin can regulate the gene expression, secretion, and proteolytic activity of MMP-3, by directly acting on articular chondrocytes.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.21 no.2
• /
• pp.71-79
• /
• 1988

For the purpose of producing low salt fermented anchovy by accelerated method with a strong proteolytic bacteria, in this study, a strong proteolytic bacterium was isolated from low salt fermented anchovy and its bacteriological characteristics and properties of protease were experimented. The results obtained were as fellows : three proteolytic bacteria, Aeromonas anaerogenes Barillus subtilis and Staphylococcus saprophyticus were isolated from low salt fermented anchovy($4\%\;of\;salt,\;4\%\;of\;KCl,\;0.5\%\;of\;lactic\;acid,\;6\%$of sorbitol and $4\%$ of alcohol extract of red pepper) after 40 days fermentation. Among these strains, which grow best at $30^{\circ}C$, pH 7.0, B. subtilis was found the best proteolytic strain and benefit for industrial use as shown $0.95\;hr^{-1}$ of specific growth rate, $89{\mu}g-Tyr/hr.ml$ of maximum activity after 12 hrs culture in TPY broth. The protease produced by by B. subtilis showed maximum activity at $35^{\circ}C$, pH 7.0, and molecular weight was estimated to be 23,000 by Sephadex G-100 filtration, and it was supposed to be a kind of metal chelator sensitive neutral protease from the results of strong sensitivity against EDTA, o-phenanthroline and metal ions such as $Cu^{2+},\;Ni^{2+},\;Fe^{2+}.Km$ value of that by method of Lineweaver-Burk was determinded to be $0.73\%$ for casein as a substrate.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.45 no.9
• /
• pp.1316-1323
• /
• 2016

The aim of this study was to evaluate proteolytic and antioxidant activities of fresh pineapple and kiwi juices extracted using a slow-speed masticating household juicer during low temperature storage. While over 90% of vitamin C and total polyphenols in both juices were retained after storage for 30 days at $-20^{\circ}C$, reduction of 56.8% for vitamin C and 31.9% for total polyphenols in pineapple juice were detected after storage at $4^{\circ}C$. In the case of kiwi juice, 32.9% of vitamin C and 22.4% of total polyphenols were lost. A high initial content of vitamin C in kiwi juice resulted in a slower reduction rate than that for pineapple juice. A similar result was obtained for 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Proteolytic activities of both juices were maintained efficiently with less than 10% loss during storage for 30 days at $-20^{\circ}C$. Protease stability of pineapple juice was better than that of kiwi juice during storage at $4^{\circ}C$, and the same result was obtained when boiled chicken breast was used as a substrate. From these results, when storing pineapple and kiwi juices, which are widely used as a natural meat tenderizer and digestive aid, cold storage at $-20^{\circ}C$ seemed to be more suitable for maintaining antioxidant and proteolytic activities than cold storage at $4^{\circ}C$.

• Korean Journal of Food Science and Technology
• /
• v.34 no.5
• /
• pp.805-810
• /
• 2002

Development of tenderizer using domestic fruits was studied. Kiwifruit was dried using various methods, and the quality of kiwifruit powder was observed during 12 week storage. Frozen kiwifruit was prepared in paste, dice, and whole flesh. After drying, paste-type kiwifruit showed 2.0 and 1.3 times higher proteolytic activity than dice and whole flesh kiwifruits, respectively. Nine hour of hot-air drying or 46 h of freeze-drying eliminated more than 90% of water from kiwifruit, during which discoloring of kiwifruit occurred. Freeze-dried powder showed 6.6 times higher yield and proteolytic activity, and resulted in almost no discolorization than those of air-dried powder. Addition of bulking agent affected the quality of hot air-dried kiwifruit powder, except color, resulting in $3.2{\sim}3.6$ times higher proteolytic activity than that without bulking agent, which is comparable to 60% of the initial freeze-dried powder content. Moisture content of kiwifruit powder with bulking agent sustained consistently during 12 week storage, whereas proteolytic activity decreased for the first 4 weeks. Freeze-drying is a preferable method to produce kiwifruit powder for tenderizer, although hot air-drying with bulking agent treatment is more economical.

• Korean Journal of Microbiology
• /
• v.45 no.4
• /
• pp.291-299
• /
• 2009

To elucidate HtrA3's functional roles in the HtrA3 mediated cellular processes, it is necessary to investigate its biochemical characteristics. In the present study, we constructed the plasmids encoding putative mature HtrA3 proteins (M1-HtrA3 and M2-HtrA3) based on the putative maturation sites of highly homologous HtrA1 and mouse HtrA3. We used the pGEX bacterial expression system to develop a simple and rapid purification for the recombinant HtrA3 protein. Although yields of the mature HtrA3 proteins were slightly low as 10~50 ${\mu}g$/L, the amounts and purity of M1- and M2-HtrA3 were enough to investigate their proteolytic activities. The putative mature HtrA3 proteins have proteolytic activity which could cleave $\beta$-casein as an exogenous substrate. We compared the proteolytic activity between the HtrA family, HtrA1, HtrA2, and HtrA3. The cleavage activity of HtrA3 and HtrA2 were 2 folds higher than that of HtrA1, respectively. Our study provides a method for generating useful reagents to identify natural substrates of HtrA3 in the further studies.