• The Korean Journal of Food And Nutrition
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• v.10 no.2
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• pp.193-200
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• 1997

In order to manufacture the low salt and functional Kochujang, salt amount was reduced to 6% and chitosan was added to 0.25% to the Kochujang preparation. The contents of ash, moisture, crude fat and crude protein in Kochujang were not affected by the reduced salt concentration and chitosan addition. pH and titratable acidity were not significantly changed by the addition of chitosan. Ethanol content was higher in 6% salt Kochujang tan in 9% salt Kochujang and decreased by the addition of chitosan. Reducing sugar content was lower in 6% salt Kochujang than in 9% salt Kochujang and increased by chitosan addition. $\alpha$-Amylase activity was slightly inhibited by the addition of chitosan, however, $\beta$-amylase, acidic protease and neutral protease activities were not affected. Amino nitrogen and ammonia nitrogen contents were higher in 6% salt Kochujang than in 9% salt Kochujang, but ammonia nitrogen production was significantly decreased by chitosan addition. Also the growth of bacteria and yeasts were slightly inhibited by the addition of chitosan. From the above results we concluded that 0.25% chitosan was the good concentration to prepare the low salt and functional Kochujang.

• 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 Microbiology and Biotechnology
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• v.31 no.7
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• pp.1028-1034
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• 2021

The effect of medium composition on enzyme and β-glucan production by Aspergillus oryzae KCCM 12698 was investigated. Brown rice, rice bran, nitrogen, and ascorbic acid are key components of the synthetic medium used in liquid-state fermentation. To determine the optimal concentrations of these components for enzyme and β-glucan production, we conducted one factor at a time experiments, which showed that the optimal concentrations were 30 g/l brown rice, 30 g/l rice bran, 10 g/l soytone, and 3 g/l ascorbic acid. Pretreatment of brown rice for 60 min prior to inoculation enhanced fungal biomass, while increasing the production of enzymes and β-glucan using solid-state fermentation. Maximum fungal biomass of 0.76 mg/g, amylase (26,551.03 U/g), protease (1,340.50 U/g), and β-glucan at 9.34% (w/w) were obtained during fermentation. Therefore, solid-state fermentation of brown rice is a process that could enhance yield and overall production of enzymes and β-glucan for use in various applications.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1447-1451
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• 2004

An experiment was conducted to determine the effects of different dietary crude protein (CP) levels on growth performance, nutrient utilization, small intestine protease activity and immunity index of weaner to 2 month-old New Zealand rabbits. Eighty weaner rabbits were allocated in individual cages to five treatments in which they were fed diets with CP at 14%, 16%, 18%, 20% and 22%, respectively. The growth performance and nutrient digestibility of rabbits increased firstly when dietary CP increased, then decreased. The average daily gain was the highest and feed conversion rate was the lowest when dietary CP reached 20%, namely 34.9 g/d and 2.74:1, respectively. Maximum CP digestibility was 72.1% in the 18% CP group, maximum crude fiber digestibility of 28.4% occurred in the 16% CP group and was significantly different from other treatments (p<0.01), apparent digestibility of Lys and Val followed the same trend as CP digestibility, and reached their maximum when dietary CP was 18%. Apparent digestibility of Cys, Tyr, Leu and Thr also had a similar trend to CP digestibility. Nitrogen retention (RN) increased with CP level (p>0.05), and was highest for 20% CP treatment (1.5 g/d). The effect of CP level on the rate of digestible nitrogen (DN) converted RN was small. The spleen index, thymus index, chymotrypsin and trypsin activities in small intestine were highest when dietary CP was 16%, which were 1.0, 2.8, 15.7 U/g and 125.7 U/g, respectively. There was no significant difference among treatments (p>0.05). According to the above results, the appropriate dietary CP level from weaner to 2 month-old meat rabbits was 18-20%.

• IMMUNE NETWORK
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• v.5 no.4
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• pp.193-198
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• 2005

Background: Protease-activated receptor 2 (PAR2) belongs to a family of G protein coupled receptors activated by proteolytic cleavage. Trypsin-like serine proteases interact with PAR2 expressed by a variety of tissues and immune cells. The aim of our study was to investigate whether PAR2 stimulation can lead to the activation of human mac rophages. Methods: PAR2-mediated proliferation of human macrophage cell line THP-1 was measured with MTT assay. We also examined the extracellular regulated kinase (ERK) phosphorylation and cytokine production induced by trypsin and PAR2-agonist using western blot and enzyme-linked immunosorbent assay (ELISA), respectively. Results: Treatment of trypsin or PAR2-activating peptide increased cell proliferation in a dose-dependent manner, and induced the activation of ERK1/2 in THP-1 cells. In addition, trypsin-induced cell proliferation was inhibited by pretreatment of an ERK inhibitor (pD98059) or trypsin inhibitor (SBTI). Moreover, PAR2 activation by trypsin increased the secretion of TNF-${\alpha}$ in THP-1 cells. Conclusion: There results suggest that P AR2 activation by trypsin-like serine proteases can induce cell proliferation through the activation of ERK in human macrophage and that PAR2 may playa crucial role in the cell proliferation and cytokine secretion induced by trypsin-like serine proteases.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.12
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• pp.1822-1829
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• 2003

Porcine colostrum and milk were separated into the acid-soluble and casein fractions by acidification followed by centrifuge. The acid-soluble fraction of porcine colostrum was further separated by liquid chromatography and anisotropic membrane filtration. Trypsin and chymotrypsin inhibitory capacity in porcine colostrum, milk and their components was determined by incubating bovine trypsin or chymotrypsin in a medium containing their corresponding substrates with or without addition of various amounts of porcine colostrum, porcine milk or their components. The inhibition of insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF) degradation in pig small intestinal contents by porcine colostrum was measured by incubating iodinated IGF-I or EGF with the intestinal contents with or without addition of porcine colostrum. Degradation of labeled IGF-I or EGF was determined by monitoring the generation of radioactivity soluble in 30% trichloroacetic acid (TCA). The results showed that porcine colostrum had high levels of trypsin and chymotrypsin inhibitory activity and increased the stability of IGF-I and EGF in pig intestinal contents. The inhibitory activity declined rapidly during lactation. It was also found that trypsin and chymotrypsin inhibitory activity and the inhibition on IGF-I and EGF degradation in the acid-soluble fraction were higher than that in the casein fraction. Heat-resistance study indicated that trypsin inhibitors in porcine colostrum survived heat treatments of $100^{\circ}C$ water bath for up to 10 min, but exposure to boiling water bath for 30 min significantly decreased the inhibitory activity. Compared with the trypsin inhibitors, the chymotrypsin inhibitors were more heatsensitive. Separation of the acid-soluble fraction of porcine colostrum by liquid chromatography and anisotropic membrane filtration revealed that the trypsin and chymotrypsin inhibitory capacity was mainly due to a group of small proteins with molecular weight of 10,000-50,000. In conclusion, the present study confirmed the existence of high levels of protease inhibitors in porcine colostrum, and the inhibition of porcine colostrum on degradation of milk-borne growth factors in the pig small intestinal tract was demonstrated for the first time.

• Journal of Plant Biotechnology
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• v.31 no.2
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• pp.163-167
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• 2004

The effects of sucrose concentration on the secretion of hGM-CSF, total protein and protease into the medium were investigated in transgenic tobacco cells. The dry cell weight (11.22 g/L), hGM-CSF (181.53 $\mu\textrm{g}$/L) and total protein (66.8 mg/L) were detected as highest at 30 g/L sucrose and protease activity (2660 U/L) was highest at 120 g/L sucrose after 5-day culture. However after 10-day culture, the maximum dry cell weight (28.36 g/L) was found at 60 g/L sucrose while the maximum hGM-CSF (95 $\mu\textrm{g}$/L) was appeared at 150 g/L sucrose. The total protein and protease activity was secreted as 52.28mg/L and 3430 U/L, respectively in the same culture.

• Korean Journal of Agricultural Science
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• v.40 no.3
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• pp.221-226
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• 2013

Degrees of hydrolysis by alkaline protease produced from Serratia marcescens S3-R1 is 3.95-6.30% of whey proteins during 5, 15, 30, 60, 90, 120,180, 240 min incubation at $40^{\circ}C$. Proteolytic pattern of the whey proteins showed that various low molecular weight peptides were generated during the incubation periods. The biological function of in Raw 264.7 cells treated with whey protein hydrolytic peptides, anti-inflammatory effect showed exhibit in the expression of pro-inflammatory cytokines such as TNF-${\alpha}$, IL-6, COX-2 and iNOS by PCR analysis. COX-2 and iNOS gene expression inhibited in Raw 264.7 cells on whey protein hydrolysates below 3,000 dalton. The protease from Serratia marcescens S3-R1 showed a potential in production of low molecular weight whey protein hydrolysates which could be used for industrial application.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1993-2005
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• 2016

Bacillus pumilus is one of the most characterized microorganisms that are used for high-level production of select industrial enzymes. A novel B. pumilus SCU11 strain possessing high alkaline protease activity was obtained in our previous work. The culture supernatant of this strain showed efficient dehairing capability with minimal collagen damage, indicating promising potential applications in the leather industry. In this study, the strain's extracellular proteome was identified by LC-MS/MS-based shotgun proteomic analysis, and their related secretory pathways were characterized by BLAST searches. A total of 513 proteins, including 100 actual secreted and 413 intracellular proteins, were detected in the extracellular proteome. The functions of these secreted proteins were elucidated and four complete secretory systems (Sec, Tat, Com, and ABC transporter) were proposed for B. pumilus. These data provide B. pumilus a comprehensive extracellular proteome profile, which is a valuable theoretical and applicative basis for future genetic modifications and development of industrial enzymes.