• Title/Summary/Keyword: Acid protease

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Preparation and Availability Analysis of Vegetable Glycoprotein (식물성 당단백질의 제조 및 유효성 분석)

  • Lee, Mi-Jin;Jeong, Noh-Hee
    • Journal of the Korean Applied Science and Technology
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    • v.26 no.3
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    • pp.248-262
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    • 2009
  • This study is on the feasibility of use of glycoprotein in various areas such as cosmetics and food etc. by extracting, isolating and refining glycoprotein from carrots, red ginseng extract residue, sesame and pine needles using protease(pepsin) and by analyzing general characteristics and measuring various bioactivities. The results of analysis of nutritional composition showed protein contents of glycoprotein. In the analysis of constitutive amino acids, the ratio of contents of hydroxy proline and glycine, the characteristics of glycoproteins appeared similar and the contents of glutamic acid and aspartic acid appeared higher. As a result of measurement contents of total polyphenol and flavonoid, it showed that glycoprotein had more contents generally, and the effect of bioactivity of glycoprotein appeared higher although different kinds of glycoprotein showed a little DPPH radical and nitrite scavenging ability, total antioxidant capacity by ABTS, ACE inhibitory.

Dyeability and Enzymetic Treatment of Wool Pretreated with tert-Butoxide (tert-Butoxide 처리 양모의 효소처리 및 염색성에 관한 연구)

  • 윤남식;윤성도
    • Textile Coloration and Finishing
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    • v.9 no.6
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    • pp.87-95
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    • 1997
  • Wool samples were treated by potassium tert-Butoxide(t-BuOK) in anhydrous tert-butanol to remove the bound surface lipid-layer, and the weight loss behaviors in protease solution and dyeabilities of the samples were studies. The C/N ratio of the surface of the t-BuOK-treated wool was shown to be 4.3 from XPS analysis. From SEM pictures any remarkable change in the shape of surface curticle during the proteasw treatment was not observed regardless of prior t-BuOK treatment. Dyeing rate and equilibrium adsorption of Orange II, a typical levelling type acid dye, on wool were not changed by protease or t-BuOK treatment, but those of Milling Cyanine 5R, a typical milling type acid dye, on wool were greatly enhanced by t-BuOK treatment in spite that, from alkali and urea-bisulfite solubilities, no damage on the inner part of wool fiber was expected by t-BuOK treatment.

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Protoplast Fusion of Alkaline Protease Producing Bacillus subtilis (Alkaline Protease를 생산하는 Bacillus subtilis의 원형질체 융합)

  • Choi, Yang-Mi;Lee, Tae-Kyung;Hong, Bum-Shik;Sung, Ha-Chin;Yang, Han-Chul
    • Applied Biological Chemistry
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    • v.32 no.4
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    • pp.435-440
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    • 1989
  • To improve alkaline protease producing strain by protoplast fusion, a strain of Bacillus sp. was treated with $100{\mu}g/ml$ NTG (N-methyl-N'-itro-N-nitrosoguanidine) for 45 minutes and mutants of Bacillus subtilts $Arg^-,\;Try^-,\;His^-$ and $Ade^-$ were isolated. The frequency of protoplast formation was about 99%, when teas of exponential phase were treated with $200{\mu}g/ml$ lysozyme at $42^{\circ}C$ for $10{\sim}30$ minutes. In a regeneration medium containing 0.3M sodium succinate, 2.0% polyvinylpyrrolidone, 0.5% casamino acid, 10mM $MgCI_2$ and 20mM $CaCI_2$, regeneration frequency cf the isolated Bacillus subtilis strains was 25.2%. The fusion frequency between mutant was from $2.1{\times}10^{-5}$ to $8.1{\times}10^{-5}$ under optimum condition.

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Purification and Characterization of Fibrinolytic Enzyme Produced by Bacillus subtilis K7 Isolated from Korean Traditional Soy Sauce (한국재래간장 발효균 Bacillus subtilis K7 유래의 혈전용해 Protease의 정제 및 특성)

  • Kim, Doo-Young;Lee, Eun-Tag;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.176-182
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    • 2003
  • An alkaline fibrinolytic protease-producing bacteria was isolated front Korean traditional soy sauce and identified as Bacillus subtilis K7 from the results of analyses of its morphological and physiological properties, $API^{\circledR}$, and Biolog system. The enzyme was purified by 75% ammonium sulfate fractionation, QAE-Sephadex anion and SP-Sephadex cation exchange column chromatography and Sephadex G-100 gel filtration. The specific activity of the purified enByme was 233.9 unit/mg protein and the yield of enzyme was 3.8%. The homogeneity of the purified enzyme was confirmed by polyacrylamide gel electrophoresis. Molecular mass of the enzyme was estimated about 21,500 Da by SDS-polyacrylamide get electrophoresis and gel chromatography. The optimum temperature and pH for the enzyme activity were $40^{\circ}C$ and 9.0, respectively. The enzyme was stable in a pH range of 5.0 to 12.0, and 60% of its activity was lost on heat treatment at $50^{\circ}C$ for 20 min. The activity of the purified enzyme was inhibited by the presence of $Fe^{2+},\;Ag^{2+},\;Cu6{2+}$, iodoacetate, ethylene diamine tetraacetic acid (EDTA), and trans-1,2-diaminocycloheane-N,N,N',N'-tetraacetic acid (CDTA). The results indicates that the enzyme requires a metal ion for its enzymatic activity.

Overexpression of the spr D Gene Encoding Strptomyces griseus Protease D Stimulates Actinorhodin Production in Streptomyces lividans

  • Choi, Si-Sun;Chi, Won-Jae;Lee, Jae-Hag;Kang, Sang-Soon;Jeong, Byeong-Chul;Hong, Soon-Kwang
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.305-313
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    • 2001
  • The spr D gene encoding Strptomyces griseus protease D(SGPD); a chymotrypsin-like proteae, was cloned from Strptomyces griseus IFO13350 and sequence. Most of the amino-acid sequence deduced from the nucleotide sequence is idential to that Strptomyces griseus IMRU3499 except that one amino acid has been deleted and Trp 369 has been substituted into Cys369 in the SGPD from S. griseus IFO13350 without affecting the protease activity. The spr D gene was overexpressed in Streptomyce liv-idans TK24 as a heterologous host. Various media with different compositions were also used to max-imize the productivity of SGPD inthe heterologous host. The SGPD productivity was best when the transformant S. lividans TK24 was cultivated in R2YE medium. The relative chymotrypsin activity of the culture broth measured with an artificial chromogenic substrate, N-scuccinyl-ala-ala-pro-phe-p-nitroanilide, was 16 units/ml. A high level of SGPD was also produced in YEME and SAAM medial but it was relatively lower that in R2YE medium and negligible amounts of SGPD were produced in GYE, GAE and Benedict media. The growth of S. lividans reacted the maximum level of cell mass at days 3 and 4 of the culture, but SGPD production started in the stationary phase of cell growth and kept increase in till the 10$^{th}$ day of culture in R2YE and YEME medium, but in GYE media the productivity reached maximum level at 8days of cultivation. The introduction of the spr D gene into S. lividans TK24 triggered biosyntheis of the pigmented antibiotic , actinorhodin, which implies some protease may paly a very improtant role in secondary-metabolite formation in sStreptomyces.

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Reaction Characteristics and Kinetic Analysis of Enzymatic Hydrolysis of Corn Gluten Meal Using Alkaline Protease (Alkaline Protease를 이용한 Corn Gluten Meal의 효소가수분해 반응특성 및 반응속도론적 분석)

  • 김성진;이은규남충희
    • KSBB Journal
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    • v.10 no.5
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    • pp.540-546
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    • 1995
  • Dry corn gluten meal of 70% protein content was enzymatically hydrolyzed by alkaline protease in a pH-state reactor. Such process variables as temperature, pH, and enzyme-to-substrate ratio were varied, and at each condition degree of hydrolysis was monitored and calculated. The ultimate degree of hydrolysis, which ranged between 25 and 28% based on gluten protein mass, was not significantly affected by the process variables. However, $50^{\circ}C$ and pH 9-10 appeared optimum. Kinetic analysis indicated enzyme deactivation was negligible during the hydrolysis, and the experimental data were near perfectly fitted to the model kinetic equation which was modified after neglecting enzyme deactivation term. The enzyme reaction was 1$100\times$ scaled up and basically the same hydrolysis performance was resulted. Amino acid analysis showed the hydrolyzate was relatively rich in glutamine/glutamic acid, leucine, and alanine at 19.6, 16.1, and 12.3 mole %, respectively.

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Characterization of a Serine Protease from Neungee [Sarcodon aspratus(Berk, ) S. Ito] (능이[Sarcodon aspratus(Berk, ) S. Ito]에서 분리한 단백질 가수분해 효소의 특성)

  • 엄태붕;유관성;김미경;류재수;손희숙;이태규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.1
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    • pp.35-39
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    • 1991
  • Properties of a protease purified from Neungee[Sarcodon aspratus(Berk, ) S. Ito] have been investigated. The enzyme displays a glycosylated serine protease. The enzyme is able to hydrolyze alanine glycine methionine glutamine and cysteine of N-CBZ and N-t-BOC-L-amino acid derivatibes relatively strongly but splits valine proline and isoleucine derivatives with low affinity which means the enzyme has the broad substrate spectrum toward the amino acids. Interestingly the enzyme was inhibited by bromelain inhibitor. That is the active site environ-ment of the enzyme is believed to be similar to that of bromelain However peptide mapping studies show that the two enzymes have distinct different cleavage sites.

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Changes in Enzyme Acrivities of Salted Chinese cabbage and Kimchi during Salting and Fermentation (배추의 소금절임과 김치숙성 중 효소류의 활성변화)

  • 오영애;김순동
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.3
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    • pp.404-410
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    • 1997
  • Amylase, protease, polygalacturonase and $\beta$-galactosidase activities were monitored during salting of Chinese cabbage and kimchi fermentation at 1$0^{\circ}C$. A part of enzymes in the tissue of Chinese cabbage were eluted during salting, and other remained enzymes activities were decreased in proportion to the amount of elution. But total enzyme activities were increased during salting. Amylase, protease and polygalacturonase activities decreased at the early fermentation stage but increased at the late fermentation stage. $\beta$-Galactosidase activity was continuously increased during all periods of fermentation. Enzymic actions at the early fermentation stage come from Chinese cabbge and at the late fermentation stage come from major microorganisms in kimchi fermentation. Kimchi fermentation involves the activation of the enzymes by salting; hydrolysis of micromoleculars such as polysaccarides cell wall composed polysaccarides and proteins of cell wall during early fermentation of kimchi; overripening of the kimchi caused by propagation of homofermentative lactic acid bacteria which demand autotroph.

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Binding Free Energy Simulations of the HIV-1 Protease and Hydroxyethylene Isostere Inhibitors

  • Won, Yeong Do
    • Bulletin of the Korean Chemical Society
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    • v.21 no.12
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    • pp.1207-1212
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    • 2000
  • The free energy simulation technique is used to evaluate the relative binding affinity of a set of hydroxyethylene isostere inhibitors of the HIV-1 protease. The binding reactions and an alchemical mutation construct the thermodynamic cycle, which reduces the free energy difference of the binding interactions into that of the alchemical processes. In the alchemical process, a methyl group is mutated into a hydrogen atom. Albeit the change is a small perturbation to the inhibitor-protease complex, it results in 25 fold difference in the binding constants. The simulation reproduces the experimentally measured binding affinities within 2% of the free energy difference. The protonation state of the catalytic aspartic acid residues is also investigated through the free energy simulations.

Change of Protein and Amino Acid Composition During Chungkook-Jang Fermentation Using Bacillus Licheniformis CN-115 (Bacillus licheniformis CN-115 균주를 이용한 청국장 제조 과장에 있어서 단백질 및 아미노산의 변화)

  • Seok, Yeong-Ran;Kim, Yung-Hawl;Kim, Sung;Woo, Hi-Seob;Kim, Tae-Wan;Lee, Son-Ho;Choi, Cheong
    • Applied Biological Chemistry
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    • v.37 no.2
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    • pp.65-71
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    • 1994
  • Chungkook-Jang was produced by fermenting Bacillus licheniformis CN-115. The changes of chemical composition, enzyme activity, and amino acids during the fermentation were investigated. The proximate composition was shown irregular fluctuation phenomenon during the fermentation, but only the moisture tended some reducing during the fermentation just after steaming. The content of amino nitrogen was increased radically after the 36 hours of fermentation and became the highest level at 18.072 mg/g at the 60 hours of it. In accordance with the fermentation of Chungkook-Jang, pH got to the 8.39 at 60 hours with increasing, protease activity was increased according to the fermentation and acid and neutral protease activity was reduced after being reached at the highest activity at 48 hours. The most suitable pH was 6.5 and temperature was $35^{\circ}C$ for dissolution-activated of protein in the process of fermentation of Chungkook-Jang. The content of water soluble protein and the content of salt soluble protein were increased at continuously according to the fermentation time of Chungkook-Jang the largest quantity. The molecular weight of water soluble protein of Chungkook-Jang fermented for 48 hours was about 19,000. The amino acids of water soluble protein just after steaming were totally 16 kinds and proline was amino acid and them was in series by glutamic acid and serine in that ordered. The amino acids salt soluble protein, just after steaming were totally 16 kinds and was the largest quantity phenylalanine, glutamic acid and aspartic acid and aspartic acid in that order.

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