• The Korean Journal of Food And Nutrition
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• v.23 no.3
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• pp.399-404
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• 2010

This study was conducted to determine the influence of culture conditions such as temperature, time, water content, koji-thickness, and agitation on the production of protease by Rhizopus sp. ZB9, isolated from Korean Nuruk, during the preparation of rice koji, which is used in brewing the Korean rice wines, Takju and Yakju. Rice koji was made under different culture conditions, and the proteolytic activity of each koji was tested. The temperature range suitable for the production of protease was $28~32^{\circ}C$. Based on the protease and color, 60 hours of cultivation at $28^{\circ}C$ was shown to produce optimum results. The production of protease increased in proportion to the increase in water content of steamed rice from 25% to 35%. An increase in koji-thickness induced no adverse effects on the production of protease, and agitation during cultivation showed beneficial effects.

• Applied Biological Chemistry
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• v.33 no.4
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• pp.325-329
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• 1990

This study was undertaken to determine thermodynamic characteristics of B. subtilis p-4 and B. licheniformis p-5 proteases isolated from fermented anchovy paste. $K_m$ values of two proteases for casein as a substrate were 0.38mM in p-4 protease and 0.18mM in p-5 protease, respectively. Denaturation constants($K_D$) of p-4 and p-5 proteases were $12.2{\times}10^{-5}/sec\;and\;19.0{\times}10^{-5}/sec\;at\;40^{\circ}C,\;and\;35.7{\times}10^{-5}/sec\;and\;46.3{\times}10^{-5}/sec\;at\;50^{\circ}C$, respectively. Activation energies($E_a$) of p-4 and p-S pmteases were 19.6 Kcal/mole and 15.2kcal/mole, respectively. Free energy of activation(${\Delta}G^*$), activation enthalpy(${\Delta}H^*$) and activation entropy(${\Delta}S^*$) at $40^{\circ}C$ were 23.21Kcal/mole, 18.98Kcal/mole and -13.50 eu, respectively for p-4 protease and 22.93Kcal/mo1e, 14.58Kcal/mole and -26.68 eu, respectively for p-5 protease. The major amino acids in p-4 protease(151 residues of amino acid) were Gly, Glu, Pro, Asp, Ser, Ala, Lys and Leu, while those in p-5 protease(247 residues of amino acid) were Gly, Glu, Asp, Ala and Leu. It may be concluded that heat denaturation of two proteases showed liner regression curve and p-5 protease was more sensitive to heat than p-4 protease.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.73-79
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• 2010

The Korea Polar Research Institute (KOPRI) has assembled a culture collection of cold-adapted bacterial strains from both the Arctic and Antarctic. To identify excellent protease-producers among the proteolytic bacterial collection (874 strains), 78 strains were selected in advance according to their relative activities and were subsequently re-examined for their extracellular protease activity on $0.1{\times}$ ZoBell plates supplemented with 1% skim milk at various temperatures. This rapid and direct screening method permitted the selection of a small group of 15 cold-adapted bacterial strains, belonging to either the genus Pseudoalteromonas (13 strains) or Flavobacterium (2 strains), that showed proteolytic activities at temperatures ranging between $5-15^{\circ}C$. The cold-active proteases from these strains were classified into four categories (serine protease, aspartic protease, cysteine protease, and metalloprotease) according to the extent of enzymatic inhibition by a class-specific protease inhibitor. Since highly active and/or cold-adapted proteases have the potential for industrial or commercial enzyme development, the protease-producing bacteria selected in this work will be studied as a valuable natural source of new proteases. Our results also highlight the relevance of the Antarctic for the isolation of protease-producing bacteria active at low temperatures.

• Journal of the Korean Society of Clothing and Textiles
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• v.24 no.2
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• pp.205-213
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• 2000

This study investigated the effect of mixing protease and lipase on detergency. The detergency of protein soiled, oil soiled and protein-oil soiled cloths and the relative hydrolytic activity of enzymes were examined. The protease-lipase added detergent solution was most effective for the removal of protein in protein-oil soiled cloths. This is because the lipase removed the protein that was physically bound to oil as well as the protease removed the protein. The protease added detergent solution was second effective, the lipase added detergent solution was third effective, and the detergent solution without protease and lipase was the least effective. The protease-lipase added detergent solution was also most effective in the oil removal from protein-oil soiled cloths. Unlike in protein removal, however, the protease added detergent solution was more effective in oil removal than the lipase added detergent solution. This is because the removal of oil bound to protein by protease was more effective than the removal of oil by lipase. In soiling-washing cycles, however, the effects of lipase increased, and as a result, the detergency of protease added detergent solution and the lipase added detergent solution became similar.

• Korean Journal of Microbiology
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• v.46 no.3
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• pp.240-247
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• 2010

Pseudomonas aeruginosa is a Gram (-) opportunistic human pathogen causing a wide variety of infections on lung, urinary tract, eyes, and burn wound sites and quorum sensing (QS), a cell density-sensing mechanism plays an essential role in Pseudomonas pathogenesis. In order to investigate the importance of QS in the Pseudomonas infections of Korean patients, we isolated 189 clinical strains of P. aeruginosa from the patients in Pusan Paik Hospital, Busan, South Korea. The QS signal production of these clinical isolates was measured by signal diffusion assay on solid media using reporter strains. While most clinical strains (79.4%) produced the QS signals as similar level as a wild type strain, PAO1 did, where LasR, the initial QS signal sensor-regulator was fully activated, a minority of them (4.2%) produced much less QS signals at the level to which LasR failed to respond. Similarly, while 72.5% of the clinical isolates produced QS signals enough to activate QscR, an another QS signal sensor-regulator, some few of them (9%) produced the QS signals at much lower level where QscR was not activated. For further analysis, we selected 74 clinical strains that were obtained from the patients under suspicion of Pseudomonas infection and investigated the total protease activity that is considered important for virulence. Interestingly, significant portion of them showed very low protease activity (44.6%) or no detectable protease activity (12.2%). When the biofilm-forming ability that is considered very important in chronic infection was examined, most isolates showed lower biofilm-forming activity than PAO1. Similarly, significant portion of clinical isolates showed reduced motility (reduced swarming activity in 51.4% and reduced twitching activity in 41.9%), or non-detectable motility (swarming-negative in 28.4% and twitching-negative in 28.4%). Our result showed that the clinical isolates that produced QS signals at the similar level to wild type could have significantly reduced activities in the protease production, biofilm formation, and motility, and some clinical isolates had unique patterns of motility, biofilm formation, and protease production that are not correlated to their QS activity.

• The Journal of the Convergence on Culture Technology
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• v.8 no.2
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• pp.399-408
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• 2022

As part of research on the development of functional materials for antlers, lactic acid fermentation of antler extract was performed. It was intended to develop a functional material with enhanced total polyphenol and flavonoid content and enhanced antioxidant activity. During the fermentation of lactic acid bacteria, the number of proliferation, total polyphenol and total flavonoid content, DPPH radical scavenging and antioxidant activity were quantified and evaluated. As a result of adding these four types of lactic acid bacteria to the antler water extract substrate, the number of lactic acid bacteria measured was 2.04~5.00×10⁷. Meanwhile, a protease (Baciullus amyloliquefaciens culture: Maxazyme NNP DS) was added to the antler extract to decompose the peptide bonds of the contained proteins. Then, these four types of lactic acid bacteria were added and the number of lactic acid bacteria increased to 2.84×10⁷~2.21×10⁸ as the result of culture. The total polyphenol contents were 4.82~6.26g/mL in the lactic acid bacteria fermentation extracts, and after the reaction of protease enzyme and lactic fermentation, increased to 14.27~20.58 g/mL. The total flavonoid contents were 1.52~2.21 g/ml in the lactic acid bacteria fermentation extracts, and after the protease reaction and fermentation, increased to 5.59~8.11 mg/mL. DPPH radical scavenging activities of lactic acid bacteria fermentation extracts was 17.03~22.75%, but after the protease reaction and fermentation, remarkably increased to 32.82~42.90%.

• Journal of Korean Ophthalmic Optics Society
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• v.19 no.1
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• pp.51-57
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• 2014

Purpose: The present study was conducted to establish the experimental condition for the proper evaluation of protein removal efficacy when developing protein removal agents. Its protein removal efficacy was further analyzed and compared with the result from protein removal efficacy against protein deposition on contact lens to suggest the evaluation method for efficacy of protein removal agents. Methods: Protein digestibility assay presented in the Korean pharmacopoeia was selected to establish the evaluation method for efficacy of papain, pancreatin, subtilisin A and protease itself as a ingredient and protein removal tablets or solution containing those enzymes and find a suitable test conditions. Furthermore, the cleaning efficacy of commercially available protein removal tablets and solution on balafilcon A lens deposited with protein artificially was measured and the correlation between two evaluation methods was further analyzed. Results: When pancreatin itself and the product containing pancreatin was evaluated by protein digestibility assay, both reached 28 IU/mg, the standard value of protein digestibility suggested by the Korean pharmacopoeia. In case of protease and subtilisin A tested with trichloroacetic acid B solution, both of them met the enzyme activity level proposed by the manufacturers when they were evaluated by protein digestibility assay however, papain and subtilisin A tested with trichloroacetic acid A solution were not reached the enzyme activity level. Among protein removal agents, three products except a product containing pancreatin did not meet the enzyme activity value specified by the manufacturer when they were evaluated by protein digestibility assay. However, actual protein removal efficacy of three products except a papain-containing product on the lens was greater than 90% protein removal. In the case of papain-containing protein removal product, its effect was not measured by protein digestibility assay however, its actual protein removal efficacy on the lens reached 73.72%. Conclusions: From the results, it was confirmed that the efficacy of protein removal agents for contact lens should be evaluated by different method according to the type of proteolytic enzyme contained. That is, the protein removal agents containing pancreatin, protease and subtilisin A can be evaluated by protein digestibility assay and protein removal efficiency evaluation and the products containing papain can be effectively evaluated by only the evaluation method for protein removal efficiency employing the lens.

• Proceedings of the Korean Society of Dyers and Finishers Conference
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• 2008.04a
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• pp.181-183
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• 2008

Four different subtilisins of protease were investigated to see their effects on the cleaning activity. The cleaning solution was formulated with various non-ionic surfactants and other additives such as propylene glycol, triethanolamine, pH balancer etc. to evaluate their effect on enzyme activity as well. Evaluation of formulated cleaning solution was carried under K0120 using pre-soiled textiles from EMPA. The results showed that the cleaning activity on soil removal was not severly influenced by surfactant but the enzyme mostly. In addition, the activity of enzymes was not much affected by the type of surfactants as long as the surfactants were non-ionic. Liquinase among the four enzymes used in this study showed the best performance on soil removal, especially blood stain.

• Microbiology and Biotechnology Letters
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• v.46 no.2
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• pp.111-113
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• 2018

In this study, purification of cold-adapted protease from Janthinobacterium sp. was investigated. First, using gradient precipitation, protease was confirmed to be deposited in the 30-80% range of ammonium sulfate. Next, DEAE-Sepharose column was used for the binding of the protease under various conditions. The optimal binding condition was found to be pH 8.5 and flow rate of 30 ml/h. Under the optimal condition, the protease was purified with 29% recovery yield. This result can be useful for the purification of other cold-adapted protein.

• Transactions of the Korean hydrogen and new energy society
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• v.28 no.6
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• pp.697-703
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• 2017

This study investigated the high-efficient process for bioethanol from barley by various condition. First, higher concentrations of ethanol could be produced without loss of yield by using reducing water consumption. This is because it could prevent to increase viscosity despite reducing water consumption. Second, the ethanol yield could be improved by using reducing particle size of biomass (increase of enzyme reactive surface). Third, The addition of protease could have a considerable effect on yield of fermentation, which provides nutrients to the yeast. This results showed that bioethanol production would provide efficient ethanol production and lower production costs.