• Parasites, Hosts and Diseases
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• v.37 no.1
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• pp.39-46
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• 1999

As a preliminary study for the explanation of pathobiology of Neodiplostomum seoulense infection. a 54 kDa protease was purified from the crude extract of adult worms by sequential chromatographic methods. The crude extract was subjected to DEAE-Sepharose Fast Flow column, and protein was eluted using 25 mM Tris-HC1 (pH 7.4) containing 0.05. 0.1, 0.2 and 0.4 M NaC1 in stepwise elution. The 0.2 M NaCl fraction was further purified by Q-Sepharose chromatography and protein was eluted using 20 mM sodium acetate (pH 6.4) containing 0.05, 0.1. 0.2 and 0.3 M NaCl, respectively. The 0.1M NaCl fraction showed a single protein band on SDS-PAGE carried out on a 7.5-15% gradient gel. The proteolytic activities of the purified enzyme were specifically inhibited by L-trans-epoxy-succinylleucylamide (4-guanidino) butane (E-64) and iodoacetic acid. The enzyme, cysteine protease. showed the maximum proteolytic activity at pH 6.0 in 0.1 M buffer, and degraded extracellular matrix proteins such as collagen and fibronectin with different activities. It is suggested that the cysteine protease may playa role in the nutrient uptake of N. seoulense from the host intestine.

• Korean journal of food and cookery science
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• v.27 no.3
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• pp.29-37
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• 2011

Protease activity of fig (Ficus carica L.), cultivated in Korea was estimated. In particular, the proteolytic effect on myofibrilar protein was studied. A crude protease extract of fig was prepared in two ways; fig was homogenized in buffer followed by centrifugation, and the supernatant was precipitated by saturated ammonium sulfate followed by dialysis. The former method resulted in 41.15 mM/g fig protease activity, whereas the latter method resulted in 17.65 mM/g fig protease activity. The crude fig protease extract showed high specificity for casein as a substrate followed by egg white, bovine serum albumin, myofibrilar protein, collagen, and elastin. The extract had stable proteolytic activity in a pH range of 6.5~9.0 (optimal at pH 7-8) but lost activity, at pH 2-3. Proteolytic activity for myofibrilar protein was sensitive to pH. The proteolytic activity of the fig extract was steady up to $60^{\circ}C$ but declined at higher temperature. It also began to lose stability in salt concentrations >0.7 M NaCl. Fig has been used as a meat tenderizer for cooking, and these results support the tenderizing effectiveness of fig, particularly for Korean style meat marinating.

• Korean Journal of Food Science and Technology
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• v.42 no.5
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• pp.554-558
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• 2010

In the study, the protease activity of kiwifruit (Actinidia deliciosa Planch) cultivated in Korea was estimated, with specific examination of proteolytic effects on myofibrilar protein. The crude protease extract of kiwifruit was prepared in two ways; one in which the kiwifruit was homogenized with buffer followed by centrifugation, and the other were the supernatant was precipitated by saturated ammonium sulfate followed by dialysis. The former had 21.23 mM/mL of protease activity, which corresponded to 112.28 mM/g kiwifruit utilized, and the latter had 11.58 mM/mL and 45.80 mM/g of kiwifruit. The crude protease extract of the kiwifruit showed high specificity for casein substrate followed by bovine serum albumin, egg white, collagen, and elastin, in order. The enzyme lost proteolytic activity in acidic conditions such as pH 2-3, and at high temperatures over $60^{\circ}C$. It showed optimal activity in both pH 3.0 and pH 7.5 as well as at $40^{\circ}C$ for casein substrate and at $50^{\circ}C$ for myofibrilar protein substrate. The proteolytic activity toward casein was high with up to 0.5M salt, followed by a sharp decrease beyond this concentration. On the other hand the proteolytic activity for myofibrilar protein decreased steadily with increasing of salt concentration. Kiwifruit has been used as a for meat tenderizer for in home cooking and these results support the its tenderizing effectiveness of kiwifruit especially for Korean style marinating of meat for cooking.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.89-94
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• 2011

Objective : Kyenegum has been frequently used for characterizing digestive symptoms in the traditional and oriental medicines. This study was conducted to investigate the characteristics of extracts from velvet antlers using the 4 different kinds of extracting methods. Methods : The extracts of velvet antlers were extracted using a $65^{\circ}C$ DW (9hrs), a Kyenegum crude enzyme, a $121^{\circ}C$ DW (2hrs), and a Kyenegum protease. To evaluate the characteristic of velvet antler extracts, we examined the brix, soluble solid, amino acid, mineral composition, and collagen protein. Results : As a result of the comparisons of velvet antlers extracted by the traditional extraction and the crude enzyme of kyenegum, the brix and soluble solid showed the higher contents for kyenegum enzymes. Also, mineral contents of the extracted velvet antlers were higher, particularly in Ca and P for those. The contents of collagen protein, hydroxyproline and hydroxylysine, were found to be more than twice in kyenegum protease compared with other extracting methods. Conclusion : These results indicated that the Kyenegum crude enzyme and protease are very effective to extract of velvet antlers.

• Korean Journal of Microbiology
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• v.51 no.1
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• pp.75-80
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• 2015

Pseudoalteromonas donghaensis HJ51, isolated from the East Sea, has been reported as a novel strain to produce extracellular protease. Crude supernatant was used to determine optimal activity and optimal production conditions for the enzyme. It was found that the optimal temperature and pH of the protease were $40^{\circ}C$ and pH 7.5-10.5, respectively. The enzyme activity was kept to 88% at the pH 11. In metal requirement analysis, the enzyme exhibited the highest activity when 10 mM $Fe^{3+}$ was supplied. While supplementation of additional carbon sources used in study showed no positive effect on cell growth and enzyme activity, the addition of beef extract, tryptone, or casamino acids instead of peptone of PY-ASW containing 1% glucose increased enzyme production to 21, 7, 4%, respectively. Taken together these properties, the enzyme produced from P. donghaensis HJ51 can be applied to the industries that require protease activity under alkaline pH and low temperature.

• Parasites, Hosts and Diseases
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• v.30 no.3
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• pp.227-230
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• 1992

Spirometra mansoni plerocercoid (sparganum) was incubated in saline at $4^{\circ}C{\;}or{\;}37^{\circ}C$ up to 100 hours. Protein contests in the excretory$.$secretory product (ESP) were rather constant (mean 7.7 mg of protein/gram of sparganum) in the preparations. Reducing SDS-PAGE of ESP showed similar protein subunit compositions with those in crude extract. Antigenic 36 and 31 kDa Proteins were major bands in ESP. ESP exhibited specific activities of protease(2.9~5.3 units/mg) at pH 6.0 and pH 7.5. Presence of protease activity in ESP may be a supporting evidence that hitherto known cysteiRe protease of sparganum is possibly secreted.

• Korean journal of food and cookery science
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• v.26 no.3
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• pp.323-329
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• 2010

Studies on the tenderizing effect of fruits has been limited even though fig, kiwifruit, pear, and pineapple cultivated in Korea are utilized commonly during cooking for their proteolytic properties. Therefore, the characteristics of these fruits were investigated by treating beef with their crude protease extracts. The protease effects of crude protease extract from the fruits on casein and myofibrilar protein were in the following order : pineapple > kiwifruit > fig > pear. Electrophoretic analysis results found that pineapple, kiwifruit, and fig cleaved myosin heavy chain into smaller fragments. The myofibrilar fragmentation ratio of crude protease extracts was the highest for pineapple whileas the lowest for pear. Ground fruits (5% and 10%) increased amounts of soluble nitrogen and decreased shear force of beef. Pineapple was the most effective while pear was the least effective. Decrease in springiness and gumminess was observed by texture profile analysis of beef treated with fruits, especially pineapple and kiwifruit. Among the 5% treatments, pineapple and kiwifruit produced the highest tenderness. Additionally, 10% treatment was less preferable than the 5% treatment.

• KSBB Journal
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• v.16 no.2
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• pp.158-162
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• 2001

A halotolerant and extracellular protease-producing yeast was isolated from traditional Meju and identified as a strain of Hansenular polymorpha by investigating its microbiological characteristics. The optimum pH, temperature and NaCl concentration reauired for the growth of Hansenular polymorpha S-9 were found to be pH 6.0, 30$^{\circ}C$ and 0.5 M, respectively. Extracellular protease was produced maximally at 10 U ml(sup)-1 when Hansenular polymorpha S-9 was grown on the medium containing 1.0% beef extract and 0.1 M NaCl for 12 hr at 30$^{\circ}C$. About 13% of the angiotensin-converting enzyme (ACE) inhibitory activity was shown in the hydrolysates which were obtained from the digestion of soybean protein (6 mg) for 6 hr at 30$^{\circ}C$ by the crude enzyme (1 U).

• Korean Journal of Microbiology
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• v.40 no.3
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• pp.230-231
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• 2004

The mycelia of Sparassis crispa DSMZ 5201 were cultivated at $24^{\circ}C$ for 15 days in yeast-malt extract-glucose broth (pH 4.0) and the filtrate was used as crude enzyme solution to determined the extracellular enzyme activity. The specific activity of $\alpha$-amylase was 44.27 unit/protein. The specific activities of protease, CMCase, $\beta$-glucosidase, chitinase, exo-$\beta$-l,4-glucanase were relatively high. However, a very little activity of xylanase was found.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.161-161
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• 1998

To achieve the aim of this investigation, the extracellular protease was isolated from bacteria BAC-4, a strain was cultivated in the medium for the production of penicillin acilase in a period of 32 hours. The enzyme was first purified by aceton precipitation method, followed by ion exchange chromatography on DEAE-sephacel column. The highest specific activity of the aceton fraction was found to be 2.19 unit per mg, with degree of purification of 13 times. Further purification of the enzyme on DEAE -sephacel had a specific activity of 58.6 unit per mg and degree of purification of 344 times compared to its crude extract. The optimum pH of the enzyme was 8.4, and the potimum temparature was 37$^{\circ}C$. The K$\_$M/ and $V_{max}$ calculated at experiment conditions were found to be 0.66%(W/V) and 3.61 unit per mL respectively.