• Title/Summary/Keyword: Acid protease

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Synthesis and Evaluation of Copoly(chitosan-g-L-lysine) for Wound Covering Material (Chitosan과 L-lysine의 공중합체로 부터 인공피부의 제조와 평가)

  • Kim, K.Y.;Min, D.S.;Park, S.H.;Lee, S.Y.;Cho, Y.J.;Chung, Y.H.;Kim, J.M.
    • Proceedings of the KOSOMBE Conference
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    • v.1989 no.05
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    • pp.7-8
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    • 1989
  • The graft copolymer of chitosan with amino acid, L-lysine was synthesized by heterogeneous copolymerization and was evaluated as an artificial skin. The mechanical properties under dry and wet state, water content, water vapor transmittance rate and biodegradability were measured. The tensile strength and elongation under wet state ranged $0.3-0.5\;kg/mm^2$, 10-13%, respectively. Water vapor transmittance rate ranged $450-500\;g/m^2{\cdot}day$ like that of the normal skin. The weight loss of prepared membrane by protease IV was measured for the degree of biodegradation. The degree of biodegradation was around 15% and after 4 days it was slow. Biocompatibility was evaluated by studying the attachment of human fibroblast on the prepared membrane surface.

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Characterization and Expression in Escherichi coli of Streptococcus pneumoniae FtsH

  • Kim, Hee-Soo;Lee, Jae-Jung
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.109-115
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    • 2000
  • FtsH is a membrane-bound, ATP-dependent metalloprotease that is involved in a variety of cellular functions including the regulation of responses to heat and stress shock. Previously, we had cloned and sequenced pneumococcal ftsH gene whose deduced amino acid sequence was very similar to those of several gram-positive bacteria and Escherichia coli, except for the N-terminal domain that was responsible for membrane anchoring. In order to better understand the role of Streptococcus pneumoniae FtsH, we expressed pneumococcal ftsH gene in Escherichia coli. When it was expressed from a strong promoter, $P_{tac}$, a considerable amount of the recombinant FtsH was produced, although the prolonged induction resulted in not only accumulation of breakdown products but also ceasing of the further growth of E. coli host. This indicated that the expression of the exogenous ftsH gene was tightly regulated since the excessive FtsH appeared detrimental to bacterial cells. In Western blotting, the pneumococcal FtsH protein, whether native or recombinant, was reactive to anti-E. coli FtsH serum. The observation that FtsH proteins were well conserved throughout the bacterial kingdom and its expression level was fine-tuned suggests an important role for this protein in the stress adaptation which may be related to infecting process by pneumococci.

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Inhibitory Effect of Lactobacillus plantarum K11 on the Adhesion of Escherichia coli O157 to Caco-2 Cells

  • Lim, Sung-Mee;Ahn, Dong-Hyun;Im, Dong-Soon
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.343-349
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    • 2009
  • Inhibitory effect of Escherichia coli O157 adhered to Caco-2 cells by the cells of Lactobacillus plantarum K11 and the cell-free culture supernatant (CFCS) and bacteriocin prepared from this strain was investigated. As the cell counts of viable L. plantarum K11 previously adhered to Caco-2 were increased, the rate of adhesion and adherent cell counts of E. coli O157 was lower. However, because the heated L. plantarum K11 rarely have the adhesion ability to Caco-2, the adhesion rate and adherent cell counts of E. coli O157 were high. In addition, the inhibitory effects of E. coli O157 adhesion by the CFCS and bacteriocin of L. plantarum K11 were dose-dependent manner. Therefore, the inhibition of adhesion of E. coli O157 to Caco-2 may result from the antimicrobial substances such as lactic acid and bacteriocin. Moreover the inhibitory activity of adhesion by the heated bacteriocin for 30 min at 100oC was similar to activity of non-treated bacteriocin, but the activity was disappeared by treatment with protease.

Analysis of the Taste Components and Antioxidant Properties of Cheonggukjang Containing Korean Red Ginseng

  • Kim, Eun-Jung;Hong, Ju-Yeon;Shin, Seung-Ryeul;Moon, Yong-Sun;Yoon, Kyung-Young
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.53-59
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    • 2009
  • This study was performed to investigate the taste composition and antioxidant properties of cheonggukjang containing Korean red ginseng (RGC), as compared to either general cheonggukjang (GC) or non-fermented boiled soybeans (BS). Amylase activity was the highest (576.7 unit/g) in RGC, whereas protease activity was the highest (326.0 unit/g) in GC. The total soluble sugar contents of BS, GC, and RGC were 2,027.5, 905.5, and 837.5 mg/100 g, respectively. RGC had the highest amount of total amino acids (2,127.4 mg/100 g) and essential amino acid (50.9%) among the samples. The ratio of sweet to bitter components was higher in RGC than in GC. Although the extracts of RGC had higher radical scavenging activity for 1,1-diphenyl-2-picrylhydrazyl (DPPH) than BS or GC, regardless of the extract concentration, the ethanol extract of RGC showed the highest scavenging ability (92.4%) at 2.0 mg/mL. The chloroform extracts from GC and RGC showed their greatest superoxide dimutase-like activities at 17.2 and 19.7% at a concentration of 2 mg/mL, respectively. Regardless of the samples, the nitrite scavenging ability was positively correlated to the extract concentration, and RGC had highest ability among samples under the same extract concentrations.

Influence of Extrusion on the Solubility of Defatted Soybean Flour in Enzymatic Hydrolysis

  • Cha, Jea-Yoon;Shin, Han-Seung;Cho, Yong-Jin;Kim, Chong-Tai;Kim, Chul-Jin
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.543-548
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    • 2007
  • Low-energy processing technology, which enhances the utility of defatted soybean flour (DSF), was developed using extrusion processing. DSF was extruded at different conditions using a twin screw extruder and then, dried at $40^{\circ}C$ for 20 hr. The nitrogen solubility index (NSI), viscosity, water solubility index (WSI), and water absorption index (WAI) of DSF increased after extrusion processing. The density of DSF extrudates decreased with the decrease in water content from 53 to 33% and the increase in extrusion temperature from 110 to $160^{\circ}C$. The addition of NaOH from 1.2 to 1.8% and citric acid from 1 to 5% increased the total solubility (TS) of DSF due to the decrease of protein coiling and hydrophobic bonds formation during extrusion processing. When viscozyme was reacted first, TS, NSI, and soluble carbohydrate content of DSF hydrolysates increased about 12, 6, and 7%, respectively, compared to them reacted with protease first. The TS and NSI of DSF hydrolysates were increased about 15 and 10%, respectively, by extrusion processing at alkaline and acidic pH. Extrusion processing at alkaline and acidic pH contributed the increase of efficiency to hydrolyze DSF samples using enzyme.

Characteristics of Kochujang Prepared by Monascus anka koji (홍국 코오지를 이용한 고추장의 특성)

  • 서형주;정수현;홍재훈;이효구;조원대
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.1
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    • pp.61-66
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    • 1999
  • The purpose of this study was to improve the palatability of kochujang. The activities of liquefying amylase and saccharogenic amylase in Monacus anka koji were lower than those in Asp. oryzae koji. The acid protease activity(1.4 units/g) in M. anka koji was also lower than that(1.6 units/g) in Asp. oryzae koji. Glucosamine amount in Asp. oryzae koji was 1075 g, and that in M. anka koji was 318 g. Four kinds of Kochujang were prepared with Asp. oryzae koji(A), M. anka koji(M), mixed koji of Asp. oryzae and M. anka(A+M), and mixture of M. anka koji and malt(M+M). The pH of four kinds of kochujang was 5.1 in the beginning and was between 4.70~4.83 after 120 days of fermentation. Reducing sugar of kochujang was between 16.3~20.7% after fermentation and kochujang prepared with M+M showed the highest reducing sugar content. Amino nitrogen of kochujang was between 182 mg%~230 mg% after fermentation and the highest amino nitrogen content was observed at kochujang prepared with A+M. Kochujang prepared with M and M+M showed higher a value than kochujang prepared with A and A+M.

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Effects of Guanidination with Trypsin, Lys-C, or Glu-C Digestion on Mass Spectrometric Signal Intensity and Protein Sequence Coverage

  • Han, Hye-Sun;Nho, Seon-Ho;Lee, Ae-Ra;Kim, Jeong-Kwon
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1527-1534
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    • 2010
  • The conventional peptide modification process of guanidination, in which the amino groups of lysine residues are converted to guanidino groups using O-methylisourea to create more basic homoarginine residues, is often used to improve the signal intensity of lysine-containing peptides in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Here, we used three different protease enzymes (trypsin, Lys-C, and Glu-C) to evaluate the effects of guanidination on the MS signals of two enzymatically digested proteins. Horse heart myoglobin and bovine serum albumin were guanidinated either before or after digestion with trypsin, Lys-C, or Glu-C. The resulting peptides were subjected to MALDI-MS, and signal intensities and sequence coverage were systematically evaluated for each digest. Guanidination prior to Glu-C digestion improved sequence coverage for both proteins. For myoglobin, guanidination before enzymatic digestion with trypsin or Lys-C also enhanced sequence coverage, but guanidination after enzymatic digestion enhanced sequence coverage only with Lys-C. For albumin, guanidination either before or after Glu-C digestion increased sequence coverage, whereas pre- or post-digestion guanidination decreased sequence coverage with trypsin and Lys-C. The amino acid composition of a protein appears to be the major factor determining whether guanidination will enhance its MALDI-MS sequence coverage.

Changes in Microorganisms, Enzyme Activities and Taste Components of Kochujang Added with Maesil Extract during Fermentation

  • Lee, Min-Ji;Seog, Eun-Ju;Lee, Jun-Ho
    • Preventive Nutrition and Food Science
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    • v.12 no.1
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    • pp.51-57
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    • 2007
  • The effects of maesil extract addition on the palatability and quality of kochujang, a traditional Korean condiment, were investigated in terms of the microbial characteristics, enzyme activities, and taste components during 100 days of fermentation. Viable cell counts of bacteria and yeast in maesil extract-added kochujang (maesil kochujang) were increased in proportion to added maesil extract from 1 to 5% during fermentation, up to 80 and 20 days, respectively and maintained $5.0{\sim}14.5{\times}10^6$ CFU/g. Activities of α-amylase, $\beta$-amylase, and protease were also increased in proportion to added maesil extract up to 20, 20, and 60 days, respectively and were higher than those of control during the aging process. The major organic acids in maesil kochujang were citric and malic acid. Composition and content of free sugar were not changed remarkably in general by the addition of maesil extract except maltose. The major free sugars of maesil kochujang were in the order of glucose>sucrose>maltose, and glucose content decreased significantly as the ratio of maesil extract increased, while maltose content increased significantly (p<0.05).

Stereoselective Synthesis of Diverse α-Hydroxy-β-amino Acids and It's Application for Synthesis of Dipeptide Expecting as a Protease Inhibitor

  • Jang, Sang-Hun;Kim, Ju-Young;Kim, Min-Kyu;Han, Jeong-Woo;Park, Ki-Hun;Yoon, Yong-Jin;Lee, Sang-Gyeong
    • Bulletin of the Korean Chemical Society
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    • v.30 no.1
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    • pp.163-171
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    • 2009
  • Few $\alpha$-Hydroxy-$\beta$-amino acids were synthesized via various nucleophilic addition of the epoxide and followed by stereoselective nucleophilic substitution reaction and eliminative cleavage of the acetal selectively in diacetal compound. One of the synthesized $\alpha$-Hydroxy-$\beta$-amino acid reacted with L-leucine methylester to give corresponding dipeptide in good yields.

Physiological and Molecular Characterization of a Newly Identified Entomopathogenic Bacteria, Photorhabdus temperata M1021

  • Jang, Eun-Kyung;Ullah, Ihsan;Lim, Jong-Hui;Lee, In-Jung;Kim, Jong-Guk;Shin, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1605-1612
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    • 2012
  • The present study concerned the identification and characterization of a novel bacterial strain isolated from entomopathogenic nematodes collected from different regions in Korea. The bacterial isolate M1021 was Gramnegative, bioluminescent, and produced red colonies on MacConkey agar medium. A rod-shaped structure was confirmed by the electron micrograph. Fatty acid composition was analyzed by using the Sherlock MIDI system. The identification was further supported by 16S rDNA sequence analysis, which revealed 96-99% sequence homology with strains of Photorhabdus temperata. The location of the isolated strain of P. temperata in the phylogenetic tree was confirmed and it was named P. temperata M1021. P. temperata M1021 exhibited catalase, protease, and lipase activities when grown on appropriate media supplemented with respective substrates. The culture of P. temperata M1021 exhibited insecticidal activity against the larvae of Galleria mellonella and the activity was the highest after 3-4 days of cultivation with agitating at $28^{\circ}C$ under 220 rpm. Antibacterial activity was also observed against Salmonella Typhimurium KCTC 1926 and Micrococcus luteus KACC 10488.