• Title/Summary/Keyword: protease production

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Whey protein hydrolytic properties and its immunomodulation activity by produced enzyme from Serratia marcescens S3-R1 (Serratia marcescens S3-R1이 생산한 효소에 의한 유청단백질 가수분해물의 특성과 면역조절 활성)

  • Yu, Jae Min;Renchinkhand, G.;Jeong, Seok Geun;Bae, Hyoung Churl;Nam, Myoung Soo
    • 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.

Extracellular Proteome Profiling of Bacillus pumilus SCU11 Producing Alkaline Protease for Dehairing

  • Wang, Chao;Yu, Shiqiang;Song, Ting;He, Tingting;Shao, Huanhuan;Wang, Haiyan
    • 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.

Properties of Protease from Aeromonas hydrophila AM-28 Isolated from Soil (토양에서 분리된 Aeromonas hydrophila AM-28이 생산하는 단백질 가수분해효소의 특성)

  • Kim, In-Sook;Kim, Hyung-Kwoun;Lee, Jung-Kee;Bae, Kyung-Sook;Oh, Tae-Kwang
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.291-296
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    • 1994
  • A bacterial strain NO. AM-28, showing proteolytic activity against defatted soybean was isolated from domestic soil. The isolated strain was identified as Aeromonas hydrophila by both the biochemical tests using API kit and the analysis of cellular fatty acid profile with MIDI system. The protease production from A. hydrophila AM-28 was highly enhanced when it was cultivated in the medium containing glycerol as a carbon source, tryptone or $(NH_4)_2HPO_4$ as a nitrogen source, and $CaCl_2$ as a mineral source. The optimal pH and temperature for the enzyme was 8.0 and $65^{\circ}C$, respectively. The enzyme was stable up to $55^{\circ}C$ and at pH values ranging from 7.0 to 13.0. The enzyme activity was inhibited by phenylmethylsulfonyl fluoride and EDTA, indicating that serine residue and metal ions be involved in enzyme activity.

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Screening of Antagonistic Bacteria for Biological control of Ginseng Root Rot (인삼뿌리썩음병 방제에 유효한 길항미생물의 탐색)

  • Kim, Young-Sook;Lee, Myeong-Seok;Yeom, Ji-Hee;Song, Ja-Gyeong;Lee, In-Kyoung;Yeo, Woon-Hyung;Yun, Bong-Sik
    • The Korean Journal of Mycology
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    • v.40 no.1
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    • pp.44-48
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    • 2012
  • Ginseng (Panax ginseng) is one of the most widely cultivated medicinal herb in Korea. However, yield losses reached up to 30~60 % due to various diseases during 3 or 5 years of ginseng cultivation. Therefore, successful production of ginseng roots depends primarily on the control of diseases. The objective of this study is to select potential multifunctional biocontrol agents from actinomycetes for the control of multiple ginseng diseases as an alternative to fungicides. Ninety three Streptomyces strains were selected and their ability to produce antibiotics, siderophore and lytic enzymes such as protease and cellulose were investigated. Eight of the isolates, strains A75, A501, 515, 523, A704, A1444, A3265 and A3283 produced cellulase and protease. These strains also produced siderophore and showed potent antifungal activity against Botrytis cinerea, Cylindrocarpon destructans, Collectotricum gloeosporioides, Phytophthora capsici and Rhizoctonia solani causing ginseng root rot.

Cultivable Microbial Diversity in Domestic Bentonites and Their Hydrolytic Enzyme Production

  • Seo, Dong-Ho;Cho, Eui-Sang;Hwang, Chi Young;Yoon, Deok Jun;Chun, Jeonghye;Jang, Yujin;Nam, Young-Do;Park, So-Lim;Lim, Seong-Il;Kim, Jae-Hwan;Seo, Myung-Ji
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.125-131
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    • 2019
  • We have isolated and identified 72 bacterial strains from four bentonite samples collected at the mining areas located in Gyeongsangbuk-do, Republic of Korea, and measured their hydrolytic enzyme (${\alpha}$-amylase, protease, and cellulase) activities to identify the isolates with industrial-use potential. Most of the isolates belonged to the Bacillaceae, with minor portions being from the Paenibacillaceae, Micrococcaceae, and Bacillales Family XII at the family level. Of the strains isolated, 33 had extracellular ${\alpha}$-amylase activity, 30 strains produced cellulase, and 35 strains produced protease. Strain MBLB1268, having the highest ${\alpha}$-amylase activity, was identified as Bacillus siamensis ($0.38{\pm}0.06U/ml$). Bacillus tequilensis MBLB1223, isolated from Byi33-b, showed the highest cellulase activity ($0.26{\pm} 0.04U/ml$), whereas Bacillus wiedmannii MBLB1197, isolated from Zdb130-b, exhibited the highest protease activity ($54.99{\pm}0.78U/ml$). These findings show that diverse bacteria of the Bacillaceae family adhere to and exist in bentonite and are potential sources of industrially useful hydrolytic enzymes.

Environment-Sensitive Ectodomain Shedding of Epithin/PRSS14 Increases Metastatic Potential of Breast Cancer Cells by Producing CCL2

  • Jang, Jiyoung;Cho, Eun Hye;Cho, Youngkyung;Ganzorig, Binderya;Kim, Ki Yeon;Kim, Moon Gyo;Kim, Chungho
    • Molecules and Cells
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    • v.45 no.8
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    • pp.564-574
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    • 2022
  • Epithin/PRSS14 is a membrane serine protease that plays a key role in tumor progression. The protease exists on the cell surface until its ectodomain shedding, which releases most of the extracellular domain. Previously, we showed that the remaining portion on the membrane undergoes intramembrane proteolysis, which results in the liberation of the intracellular domain and the intracellular domain-mediated gene expression. In this study, we investigated how the intramembrane proteolysis for the nuclear function is initiated. We observed that ectodomain shedding of epithin/PRSS14 in mouse breast cancer 4T1 cells increased depending on environmental conditions and was positively correlated with invasiveness of the cells and their proinvasive cytokine production. We identified selenite as an environmental factor that can induce ectodomain shedding of the protease and increase C-C motif chemokine ligand 2 (CCL2) secretion in an epithin/PRSS14-dependent manner. Additionally, by demonstrating that the expression of the intracellular domain of epithin/PRSS14 is sufficient to induce CCL2 secretion, we established that epithin/PRSS14-dependent shedding and its subsequent intramembrane proteolysis are responsible for the metastatic conversion of 4T1 cells under these conditions. Consequently, we propose that epithin/PRSS14 can act as an environment-sensing receptor that promotes cancer metastasis by liberating the intracellular domain bearing transcriptional activity under conditions promoting ectodomain shedding.

Effects of gold and green kiwifruit juices on the physicochemical properties and tenderness of pork loin and antioxidant activity during incubation (24 h) in a pork model system

  • Haeun Kim;Koo Bok Chin
    • Animal Bioscience
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    • v.37 no.5
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    • pp.908-917
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    • 2024
  • Objective: Although pork loins is not a tough meat, they need to develop meat products with a soft texture for the elderly. This study focused on the physicochemical properties and tenderness characteristics of pork loin injected with green kiwifruit juice (GRJ) and gold kiwifruit juice (GOJ) during various incubation times. In addition, the antioxidant activities of hydrolysate derived from the hydrolysis of pork loin by kiwifruit juice protease were evaluated. Methods: The pork loin was injected with 10% and 20% GRJ and GOJ, under various incubation times (0, 4, 8, and 24 h). Then, the physicochemical properties and tenderness of pork loins were measured. 2,2- diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power were conducted to determine hydrolysate's antioxidant activities derived from pork loin's hydrolysis by kiwifruit juice protease. Results: GRJ had greater tenderizing ability than GOJ, even at the 10% addition. When kiwifruit juice was injected into pork loin, the tenderness increased with increasing incubation time. This was confirmed by the decrease in intensity of the myosin heavy chain (MHC) band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In particular, the MHC band decreased at 8 h for both 10% GRJ and 20% GOJ and at 4 h for 20% GRJ alone. The highest myofibril fragmentation index and peptide solubility were observed in pork loin treated with 20% GRJ compared to the other treatments during incubation. The 10% GRJ and 20% GOJ treatments showed similar levels of antioxidant activity of the protein hydrolysates in pork loin, and 20% GRJ showed the highest activity among the treatments. Conclusion: Kiwifruit juice had protease activity, and GRJ was more useful for tenderizing meat products than GOJ. Thus, GRJ at 10% could be a potential agent to tenderize and enrich the natural antioxidant activity through the proteolysis of pork loin.

Evaluation of Ciclopirox as a Virulence-modifying Agent Against Multidrug Resistant Pseudomonas aeruginosa Clinical Isolates from Egypt

  • Zakaria, Azza S.;Edward, Eva A.;Mohamed, Nelly M.
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.651-661
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    • 2019
  • Targeting the pathogen viability using drugs is associated with development of drug resistance due to selective pressure. Hence, there is an increased interest in developing agents that target bacterial virulence. In this study, the inhibitory effect of ciclopirox, an antifungal agent with iron chelation potential, on the microbial virulence factors was evaluated in 26 clinical MDR Pseudomonas aeruginosa isolates collected from Alexandria Main University Hospital, a tertiary hospital in Egypt. Treatment with 9 ㎍/ml ciclopirox inhibited the hemolytic activity in 70% isolates, reduced pyocyanin production, decreased protease secretion in 46% isolates, lowered twitching and swarming motility, and decreased biofilm formation by 1.5- to 4.5-fold. The quantitative real-time PCR analysis revealed that treatment with ciclopirox downregulated the expression levels of alkaline protease (aprA) and pyocyanin (phzA1). Ciclopirox is used to treat hematological malignancies and the systemic administration of ciclopirox is reported to have adequate oral absorption with a satisfactory drug safety profile. It is important to calculate the appropriate clinical dose and therapeutic index to reposition ciclopirox from a topical antifungal agent to a promising virulence-modifying agent agent against P. aeruginosa, a problematic Gram-negative pathogen.

Lectin Activity and Chemical Characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. from Gastrointestinal Mucosa of Growing Pigs

  • Gao, W.;Meng, Q.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.863-868
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    • 2004
  • Lectin activities and chemical characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. originating from the porcine cecal mucosal layer were studied based on hemagglutination assay (HA) and hemagglutination inhibition assay (HIA). Although all the bacterial strains were able to agglutinate erythrocytes of porcine or rabbit origin, much higher HA titers were consistently observed for Lactobacillus spp. than for E. coli or for Bifidobacterium spp. A remarkable reduction in HA titers occurred by the treatment of E. coli and Lactobacillus spp. with protease or trypsin and of Bifidobacterium spp. with protease, trypsin or periodate. There were no significant effects on the HA titers of the three groups of bacteria after the treatment with lipase. Hemagglutination of E. coli was strongly inhibited by D (+)-mannose and D (+)-galactose; Lactobacillus spp. by $\alpha$-L-rhamnose and methyl-$\beta$-galactopyranoside; Bifidobacterium spp. by D (+)-alactose, $\alpha$-L-rhamnose, $\alpha$-L-fucose, L (+)-arabinose, D (+)-mannose, D (-)-fructose at a relatively low concentration (1.43 to 3.75 mg/ml). These results, combined with the enhanced HA activities of the three bacterial strains by modification of rabbit erythrocytes with neuraminidase and abolished HA activity of E. coli after treatment with $\beta$-galactosidase, indicate that it might be the glycoproteinous substances surrounding the surface of the bacterial cells that are responsible for the adhesions of these microorganisms by recognizing the specific receptors on the red blood cell.

Angiotensin I-converting Enzyme Inhibitory Activities of Porcine Skeletal Muscle Proteins Following Enzyme Digestion

  • Katayama, K.;Fuchu, H.;Sakata, A.;Kawahara, S.;Yamauchi, K.;Kawamura, Y.;Muguruma, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.417-424
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    • 2003
  • Inhibitory activities against angiotensin I-converting enzyme (ACE) of enzymatic hydrolysates of porcine skeletal muscle proteins were investigated. Myosin B, myosin, actin, tropomyosin, troponin and water-soluble proteins extracted from pork loin were digested by eight kinds of proteases, including pepsin, $\alpha$-chymotrypsin, and trypsin. After digestion, hydrolysates produced from all proteins showed ACE inhibitory activities, and the peptic hydrolysate showed the strongest activity. In the case of myosin B, the molar concentration of peptic hydrolysate required to inhibit 50% of the activity increased gradually as digestion proceeded. The hydrolysates produced by sequential digestion with pepsin and $\alpha$-chymotrypsin, pepsin and trypsin or pepsin and pancreatin showed weaker activities than those by pepsin alone, suggesting that ACE inhibitory peptides from peptic digestion might lose their active sequences after digestion by the second protease. However, the hydrolysates produced by sequential digestion showed stronger activities than those by $\alpha$-chymotrypsin, trypsin or pancreatin alone. These results suggested that the hydrolysates of porcine meat were able to show ACE inhibitory activity, even if they were digested in vivo, and that pork might be a useful source of physiologically functional factors.