• Title/Summary/Keyword: Microbial Protease

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Expression of Various Pattern Recognition Receptors in Gingival Epithelial Cells

  • Shin, Ji-Eun;Ji, Suk;Choi, Young-Nim
    • International Journal of Oral Biology
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    • v.33 no.3
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    • pp.77-82
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    • 2008
  • Innate immune response is initiated by the recognition of unique microbial molecular patterns through pattern recognition receptors (PRRs). The purpose of this study is to dissect the expression of various PRRs in gingival epithelial cells of differentiated versus undifferentiated states. Differentiation of immortalized human gingival epithelial HOK-16B cells was induced by culture in the presence of high $Ca^{2+}$ at increased cell density. The expression levels of various PRRs in HOK-16B cells were examined by realtime reverse transcription polymerase chain reaction (RTPCR) and flow cytometry. In addition, the expression of human beta defensins (HBDs) was examined by real time RT-PCR and the amounts of secreted cytokines were measured by enzyme linked immunosorbent assay. In undifferentiated HOK-16B cells, NACHT-LRR-PYDcontaining protein (NALP) 2 was expressed most abundantly, and toll like receptor (TLR) 2, TLR4, nucleotide-binding oligomerization domain (NOD) 1, and NOD2 were expressed in substantial levels. However, TLR3, TLR7, TLR8, TLR9, ICE protease-activating factor (IPAF), and NALP6 were hardly expressed. In differentiated cells, the levels of NOD2, NALP2, and TLR4 were different from those in undifferentiated cells at RNA but not at protein levels. Interestingly, differentiated cells expressed the increased levels of HBD-1 and -3 but secreted reduced amount of IL-8. In conclusion, the repertoire of PRRs expressed by gingival epithelial cells is limited, and undifferentiated and differentiated cells express similar levels of PRRs.

The Impact of Proteolytic Pork Hydrolysate on Microbial, Flavor and Free Amino Acids Compounds of Yogurt

  • Lin, Jinzhong;Hua, Baozhen;Xu, Zhiping;Li, Sha;Ma, Chengjie
    • Food Science of Animal Resources
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    • v.36 no.4
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    • pp.558-565
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    • 2016
  • The aim of this study was to investigate the influence of proteolytic pork hydrolysate (PPH) on yoghurt production by Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Fresh lean pork was cut into pieces and mixed with deionized water and dealt with protease, then the resulting PPH was added to milk to investigate the effects of PPH on yoghurt production. The fermentation time, the viable cell counts, the flavor, free amino acids compounds, and sensory evaluation of yoghurt were evaluated. These results showed that PPH significantly stimulated the growth and acidification of the both bacterial strains. When the content of PPH reached 5% (w/w), the increased acidifying rate occurred, which the fermentation time was one hour less than that of the control, a time saving of up to 20% compared with the control. The viable cell counts, the total free amino acids, and the scores of taste, flavor and overall acceptability in PPH-supplemented yoghurt were higher than the control. Furthermore, the contents of some characteristic flavor compounds including acids, alcohols, aldehydes, ketones and esters were richer than the control. We concluded that the constituents of PPH such as small peptide, vitamins, and minerals together to play the stimulatory roles and result in beneficial effect for the yoghurt starter cultures growth.

Effect of Lactic Acid Producing Bacteria on the Performance of Male Crossbred Calves Fed Roughage Based Diet

  • Ramaswami, N.;Chaudhary, L.C.;Agarwal, N.;Kamra, D.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.8
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    • pp.1110-1115
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    • 2005
  • To study the effect of feeding lactic acid producing bacteria on the performance of cattle calves, twenty four, day old male crossbred cattle calves (Bos indicus${\times}$Bos taurus), were distributed into two groups of 12 animals each. The animals were fed on calf starter containing wheat bran and green berseem ad libitum and milk as per requirement upto 8 weeks of age. The diet of calves of Group 2 was supplemented with 500 ml culture of Lactobacillus acidophilus-15. Total duration of the experiment was 31 weeks. There was no significant difference in intake and digestibility of dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF) and crude protein (CP) between the groups. The rumen pH, protozoa numbers, concentration of volatile fatty acids (VFA), ammonia nitrogen ($NH_3-N$), trichloroacetic acid precipitable nitrogen (TCA-ppt N) and activity of microbial enzymes (carboxymethylcellulase, xylanase, amylase and protease) were not affected due to probiotic supplementation. Average live weight gain of the calves was improved (about 10%) and feed:gain ratio was reduced (about 5%) in the animals given Lactobacillus culture. The data indicated that crossbred calves could be reared on a diet devoid of cereal grain and addition of Lactobacillus culture in the diet resulted in an added advantage in growth performance of the animals.

Purification and Characterization of the Fibrinolytic Enzyme Produced by Bacillus subtilis KCK-7 from Chungkookjang

  • Paik, Hyun-Dong;Lee, Si-Kyung;Heo, Seok;Kim, Soo-Young;Lee, Hyung-Hoan;Kwon, Tae-Jong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.829-835
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    • 2004
  • A fibrinolytic enzyme has been found in several bacteria isolated from fermented food. This study was carried out to investigate the purification and characteristics of the fibrinolytic enzyme produced by Bacillus subtilis KCK-7 originated from Chungkookjang. The fibrinolytic enzyme was purified to homogeneity from the culture supernatant using ammonium sulfate fractionation and chromatographies on DEAE-cellulose and on Sephadex G-100. The final specific activity of the purified enzyme increased 11.0-fold, and the protein amount in the purified enzyme was about 16% of that in the culture supernatant. The molecular weight of the purified enzyme was estimated to be about 45,000 by SDS-PAGE. The optimum pH and temperature for the enzyme activity were pH 7.0 and $60^{\circ}C$, respectively. The enzyme activity was relatively stable up to $60^{\circ}C$ over the pH range of 7.0-10.0. The fibrinolytic enzyme activity increased by $Ca^{2+}$ and $Cu^{2+}$, whereas it was inhibited by $Hg^{2+}$ and $Ba^{2+}$. In addition, it was severely inhibited by PMSF and DFT. It is suggested that the purified enzyme was a serine protease for the fibrinolysis. The purified enzyme could completely hydrolyze fibrin in vitro within 8 h. Hence, it is suggested that the purified enzyme can be put into practice as an effective thrombolytic agent.

Isolation and Characterization of Siderophore-Producing Bacteria with Various Plant Growth-Promoting Abilities as a Potential Biocontrol Agent (잠재적 미생물 농약으로서 다양한 식물성장 촉진 활성을 가진 siderophore 생산 세균의 분리와 특성)

  • Choi, Seunghoon;Yoo, Ji-Yeon;Park, SungJin;Park, MinJoo;Lee, O-Mi;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.29 no.9
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    • pp.925-933
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    • 2020
  • To develop eco-friendly microbial inoculants, siderophore-producing bacteria were isolated and identified, and their production characteristics and plant growth-promoting abilities were investigated. A strain S21 was isolated from rhizosphere of Korean perilla (Perilla frutescens) and identified as Enterobacter amnigenus by phenotypic properties and 16S rRNA gene sequencing. The highest siderophore production was obtained in a medium containing 0.5% fructose, 0.1% urea, 0.5% K2HPO4 and 0.1% succinic acid. By using this improved medium, siderophore production increased by 2.5 times compared to that of basal medium. The strain S21 showed insoluble phosphate solubilizing, ammonification and antifungal activities, and also produced hydrolytic enzymes (protease and lipase), indoleacetic acid and 1-aminocyclopropane-1-carboxylate deaminase. Our data suggest that E. amnigenus S21 is a potential candidate that can be used as eco-friendly biocontrol agent and biofertilizer.

Screening of Multifunctional Bacteria with Biocontrol and Biofertilizing Effects (식물병원진균의 생물적 방제 및 생물비료 활성을 갖는 다기능 세균의 탐색)

  • Kim, Young-Sook;Lee, Myeong-Seok;Yeom, Ji-Hee;Song, Ja-Gyeong;Lee, In-Kyoung;Yun, Bong-Sik
    • The Korean Journal of Mycology
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    • v.39 no.2
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    • pp.126-130
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    • 2011
  • In the course of search for multifunctional microbial inoculants, three Bacillus strains (BS11-1,BS11-2,BS11-3) with biological control and biofertilizing effects were selected. In this study, their ability for solubilization of insoluble phosphate, production of indole-3-acetic acid (IAA), siderophore, and hydrolytic enzymes, and antagonism against phytopathogenic fungi were estimated. All strains produced IAA and siderophore depending on culture time and produced a visible clear zone on agar plate containing 0.5% carboxylmethyl cellulose as a carbon source. Also, these strains exhibited antifungal activities against phytopathogenic fungi, Botrytis cinerea, Cylindrocarpon destructans, Fusarium oxysporum, Rhizoctonia solani, and Phytophthora capsici.

Coaggregation between Porphyromonas gingivalis and Tannerella forsythia (Porphyromonas gingivalis와 Tannerella forsythia의 응집반응)

  • Um, Heung-Sik;Lee, Seok-Woo;Park, Jae-Hong;Nauman, R.K.
    • Journal of Periodontal and Implant Science
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    • v.36 no.1
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    • pp.265-272
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    • 2006
  • Dental plaque, a biofilm consisting of more than 500 different bacterial species, is an etiological agent of human periodontal disease, It is therefore important to characterize interactions among periodontopathic microorganisms in order to understand the microbial pathogenesis of periodontal disease. Previous data have suggested a synergistic effect of tow major periodontal pathogens Porphyromonas gingivalis and Tannerella forsythia in the periodontal lesion. In the present study, to better understand interaction between P. gingivalis and T. forsythia, the coaggregation activity between these bacteria was characterized. The coaggregation activity was observed by a direct visual assay by mixing equal amount (1 ${\times}$ $10^9$)of T. forsythia and P. gingivaJis cells. It was found that the first aggregates began to appear after 5-10 min, and that the large aggregates completely settled within 1 h. Electron and epifluorescence microscopic studies confirmed cell-cell contact between two bacteria. The heat treatment of P. gingivalis completely blocked the activity, suggesting an involvement of a heat-labile component of P. gingivalis in the interaction. On the other hand, heat treatment of T. forsythia significantly increased the coaggregation activity; the aggregates began to appear immediately. The coaggregation activity was inhibited by addition of protease, however carbohydrates did not inhibit the activity, suggesting that coaggregation is a protein-protein interaction. The results of this study suggest that coaggregation between P. gingivalis and T. forsythia is a result of cell-cell physical contact, and that coaggregation is mediated by a heat-labile component of P. gingivalis and T. forsythia component that can be activated on heat treatment.

Fermentation Properties of Irradiated Kochujang (방사선 조사 고추장의 발효 특성)

  • Kim, Moon-Sook;Oh, Jin-A;Kim, In-Won;Shin, Dong-Hwa;Han, Min-Su
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.934-940
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    • 1998
  • Kochujang, fermented hot pepper soybean paste, was prepared by traditional method, and irradiated with gamma ray of $^{60}Co{\;}up{\;}to{\;}15{\pm}1.5{\;}kGy$ for confirming main factor of kochujang fermentation. Nonirradiated (control) and irradiated samples kochujang were fermented at $25^{\circ}C$ and analyzed biochemical properties including enzyme activities and viable cell count during fermentation for 60 days. The total viable count in irradiated kochujang decreased to $10^4{\;}CFU/g$ which was $10^8{\;}CFU/g$ in the control. Because of a little changing enzyme activities of ${\alpha}{\cdot}{\beta}{\;}amylase$ and acid neutral protease by irradiation at the above level, amino type nitrogen which is the main quality reference of kochujang was comparable to the control. By irradiation, gas production was completely stopped which is one of biggest problems during distribution of kochujang. Total volume of gas produced at $25^{\circ}C$ from the control kochujang was 453 mL/100 g which was composed of over 90% of $CO_2$. The odor of irradiated kochujang was inferior to the control which seemed to be related to reduced microbial populations.

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Production performances and antioxidant activities of laying hens fed Aspergillus oryzae and phytase co-fermented wheat bran

  • Huang, Chung Ming;Chuang, Wen Yang;Lin, Wei Chih;Lin, Li Jen;Chang, Sheng Chang;Lee, Tzu Tai
    • Animal Bioscience
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    • v.34 no.3_spc
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    • pp.371-384
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    • 2021
  • Objective: Wheat bran (WB) was co-fermented with Aspergillus oryzae and phytase (Phy) to determine whether co-fermentation improve WB phosphorus and fiber utilization in Isa-brown layers. Methods: A total of 112 Isa brown layer were randomly divided into 7 treatments with 8 replicates per a treatment and 2 hens per a replicate. The treatments included basal diet (control), basal diet supplemented with 250 unit/kg Phy (control+Phy), diet with 10% WB (10% WB), diet with 5% WB and 250 unit/kg Phy (5% WB+Phy) diet with 10% WB and 250 unit/kg Phy (10% WB+Phy), diet with 5% fermented WB supplemented with molasses and phy (PCFWH) and 125 unit/kg Phy (5% PCFWH), and diet with 10% PCFWH (10% PCFWH). The intestinal microbial population, intestinal morphology, serum antioxidant enzyme activities, and excreta phosphorus content were assessed. Results: In PCFWH, spore counts, protease activity, xylanase activity, and ferulic acid were 8.50 log/g dry matter (DM), 190 unit/g DM, 120 unit/g DM, and 127 ㎍/g, respectively. Xylobiose and xylotriose were released in PCFWH, while they were not detectable in WB. Antioxidant capacity was also enhanced in PCFWH compared to WB. The 10% WB+Phy and 10% PCFWH groups produced higher egg mass, but hens fed 5% WB+Phy had the lowest amount of feed intake. Eggs from 10% PCFWH had better eggshell weight, eggshell strength, and eggshell thickness. Birds fed with 10% PCFWH also had higher serum superoxide dismutase and catalase activities. Compare to control, 10% PCFWH significantly reduced excreta phosphorus content. Conclusion: Diet inclusion of 10% PCFWH improved egg quality, antioxidant status, and excreta phosphorus content of laying hens.

Effects of Dietary Prebiotics and Probiotics on Growth, Immune Response, Anti-oxidant Capacity and Some Intestinal Bacterial Groups of the Red Seabream Pagrus major (사료 내 Prebiotic과 Probiotics의 첨가가 참돔(Pagrus major)의 성장, 면역력, 항산화력, 장내 미생물 조성 변화에 미치는 영향)

  • Jongho Lim;Gunho Eom;Choong Hwan Noh;Kyeong-jun Lee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.1
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    • pp.89-98
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    • 2023
  • We evaluated the effects of prebiotic (mannan oligosaccharides, Mos) and probiotic diet supplements on growth performance, innate immunity, antioxidant activity, and intestinal changes in the microbial flora of red seabream Pagrus major. A basal diet (Con) was formulated to meet the nutrient requirement of red seabream. The dietary starch in Con was replaced with 0.6% Mos, Lactobacillus plantarum, Bacillus subtilis, B. licheniformis and probiotic mixture (labeled as Mos, Pro-LP, Pro-BS, Pro-BL and Pro-Mix, respectively). We stocked 450 fish in 18 polypropylene tanks (400 L) in triplicate groups per dietary treatment. The fish were fed one of the diets twice (08:30, 18:30 h) a day for 63 days. Lysozyme activity was significantly higher in all the supplemented groups than that of the Con group. The immunoglobulin level of Pro-Mix, anti-protease activity of Pro-BL, and glutathione peroxidase and superoxide dismutase activity of Pro-BS, Pro-BL and Pro-Mix groups were significantly higher than those of the Con group. The ratio of total Vibrio/heterotrophic marine bacteria counts was significantly lower in Pro-LP, Pro-BL and Pro-Mix groups than that of the Con group. Therefore, dietary supplementation of Mos and probiotics to improves immune response and antioxidant enzyme activity and inhibits Vibrio bacteria in the intestine.