• Title/Summary/Keyword: Activity stability

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Prokaryotic Selectivity, Anti-endotoxic Activity and Protease Stability of Diastereomeric and Enantiomeric Analogs of Human Antimicrobial Peptide LL-37

  • Nan, Yong-Hai;Lee, Bong-Ju;Shin, Song-Yub
    • Bulletin of the Korean Chemical Society
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    • v.33 no.9
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    • pp.2883-2889
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    • 2012
  • LL-37 is the only antimicrobial peptide (AMP) of the human cathelicidin family. In addition to potent antimicrobial activity, LL-37 is known to have the potential to inhibit lipolysaccharide (LPS)-induced endotoxic effects. To provide the stability to proteolytic digestion and increase prokaryotic selectivity and/or anti-endotoxic activity of two Lys/Trp-substituted 19-meric antimicrobial peptides (a4-W1 and a4-W2) designed from IG-19 (residues 13-31 of LL-37), we synthesized the diastereomeric peptides (a4-W1-D and a4-W2-D) with D-amino acid substitution at positions 3, 7, 10, 13 and 17 of a4-W1 and a4-W2, respectively and the enantiomeric peptides (a4-W1-E and a4-W2-E) composed D-amino acids. The diastereomeric peptides exhibited the best prokaryotic selectivity and effective protease stability, but no or less anti-endotoxic activity. In contrast, the enantiomeric peptides had not only prokaryotic selectivity and anti-endotoxic activity but also protease stability. Our results suggest that the hydrophobicity and ${\alpha}$-helicity of the peptide is important for anti-endotoxic activity. In particular, the enantiomeric peptides showed potent anti-endotoxic and LPS-neutralizing activities comparable to that of LL-37. Taken together, both a4-W1-E and a4-W2-E holds promise as a template for the development of peptide antibiotics for the treatment of endotoxic shock and sepsis.

Effects of Drugs on the Stability of Phospholipid Liposomal Membranes (수종 약물이 리포솜 지질막의 안정성에 미치는 영향)

  • Kim, Min;Han, Suk-Kyu;Kim, Chong-Kook
    • YAKHAK HOEJI
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    • v.38 no.6
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    • pp.637-645
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    • 1994
  • The effect of various drugs on the stability of the liposomal membrane of phosphatidylcholine and cholesterol was studied, employing the fluorescence self-quenching method. Calcein was entrapped into the phospholipid small unilamellar vesicles and the leakage of the fluorescence probe was monitored on adding the drug to the system. The results of the experiments showed that phenothiazine derivatives, some potent local anesthetics and surface active agents were very effective in inducing the leakage of calcein from the liposome. The leakage-inducing activity of these drug substances has been ascribed to their surface activity and the perturbation of the liposomal membrane by these substances. On the other hand drug substance with low surface activity or without amphiphilic moieties did not show any effect or only small effect on the leakage of calcein from the liposomes. The effect of lipid concentration on the stability of the liposomes was also investigated to show that the higher concentrations of lipid more drug was required to induce the leakage. The effect of surface charges of vesicles was also studied, and the results showed that the charge on the liposomes enhanced the stability of the liposomes against the leakage-inducing activity of these drug substances.

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Enhanced Stability of Tyrosine Phenol-Lyase from Symbiobacterium toebii by DNA Shuffling

  • Kim, Jin-Ho;Song, Jae-Jun;Kim, Bong-Gyun;Sung, Moon-Hee;Lee, Sang-Chul
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.153-157
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    • 2004
  • Tyrosine phenol-lyase (TPL) is a useful enzyme for the synthesis of pharmaceutical aromatic amino acids. In the current study, sequential DNA shuffling and screening were used to enhance the stability of TPL. Twenty-thousand mutants were screened, and several improved variants were isolated. One variant named A13V, in which the $13^{th}$ amino acid alanine was substituted by valine, exhibited a higher temperature and denaturant stability than the wild-type TPL. The purified mutant TPL, A13V, retained about 60% of its activity at $76^\circ{C}$, whereas the activity of the wild-type TPL decreased to less than 20% at the same temperature. Plus, A13V exhibited about 50% activity with 3 M urea, while the wild-type TPL lost almost all its catalytic activity, indicating an increased denaturant tolerance in the mutant A13V. It is speculated that the substitution of Val for the Ala in the $\beta$-strand of the N-terminal arm was responsible for the heightened stabilization, and that the current results will contribute to further research on the structural stability of TPL.

Stability of Sweet Potato $\beta$Amylase (I) (고구마 $\beta$아밀라아제의 안정성에 관한 연구 (1))

  • 안용근;이석건
    • The Korean Journal of Food And Nutrition
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    • v.9 no.3
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    • pp.247-252
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    • 1996
  • $\beta$-Amylase was purified from sweet potato by acetone fractlonatlon, Sephadex A-50 ion exchange chromatography and Sepgadex G-200 gel chromatographyl The higher enzyme concentration was, the higher heat stability of enzyme became. After 1 hour 30 minute. At 6$0^{\circ}C$ in pH 5, enzyme under concentration of 30$\mu$l/ml lost its activity completely and over the concentration of 100$\mu$g/ml remained 25% of activity. The enzyme was stabilized at range of pH 4~10 and pH stability was increased by glycerol. Five moles of NaCl inhibited completely of the enzyme activity. SDS of 0.05% inhibited the enzyme completely after 12 hours at 37$^{\circ}C$ in pH5. One mole guanidine-HCl and 8M urea inhibited the entire enzyme after 13 hours at 37$^{\circ}C$ in pH 5.

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Stability Analysis of Bacillus stearothermopilus L1 Lipase Fused with a Cellulose-binding Domain

  • Hwang Sangpill;Ahn Ik-Sung
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.329-333
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    • 2005
  • This study was designed to investigate the stability of a lipase fused with a cellulose­binding domain (CBD) to cellulase. The fusion protein was derived from a gene cluster of a CBD fragment of a cellulase gene in Trichoderma hazianum and a lipase gene in Bacillus stearother­mophilus L1. Due to the CBD, this lipase can be immobilized to a cellulose material. Factors affecting the lipase stability were divided into the reaction-independent factors (RIF), and the re­action-dependent factors (RDF). RIF includes the reaction conditions such as pH and tempera­ture, whereas substrate limitation and product inhibition are examples of RDF. As pH 10 and $50^{\circ}C$ were found to be optimum reaction conditions for oil hydrolysis by this lipase, the stability of the free and the immobilized lipase was studied under these conditions. Avicel (microcrystal­line cellulose) was used as a support for lipase immobilization. The effects of both RIF and RDF on the enzyme activity were less for the immobilized lipase than for the free lipase. Due to the irreversible binding of CBD to Avicel and the high stability of the immobilized lipase, the enzyme activity after five times of use was over $70\%$ of the initial activity.

Effects of adding ethanol extracts from Ulmus davidiana to Yackwa base as an antioxidant during storage

  • Sim, Ki Hyeon
    • Korean Journal of Food Science and Technology
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    • v.49 no.1
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    • pp.63-71
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    • 2017
  • This study aimed to assess the antioxidative potential of adding ethanolic extracts from Ulmus davidiana to Yackwa (Korean fried cookie). The reducing power and antioxidant activity of U. davidiana-treated Yackwa were assessed against DPPH, nitric oxide, superoxide anions, superoxide dismutase, and ABTS radicals. Increasing the amount of U. davidiana extract in Yackwa dough resulted in improved antioxidant properties. U. davidiana extracts were effective antioxidants with a radical scavenging potency similar to the potency of butylated hydroxytoluene (BHT) and L-ascorbic acid. The DPPH and nitric oxide radical scavenging activity of Yackwa containing 0.2% U. davidiana resulted in increased oxidative stability relative to control dough. Although the antioxidant effects of Yackwa containing U. davidiana extract were lower than those of Yackwa containing BHT and L-ascorbic acid, these results suggest that U. davidiana extracts have potent antioxidant activity. Accordingly, U. davidiana extract is a potential additive for deep-fried foods, such as Yackwa, to improve their oxidative stability.

On the CMCase Activity from Two species of Trichosporon (Trichosporon의 CMCase 활성에 관하여)

  • 전순배;박종영
    • Korean Journal of Microbiology
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    • v.17 no.4
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    • pp.187-192
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    • 1979
  • Dennis (1972) reported that Trichosporon cutaneum FRI-425 from the petioles of Pheum rhamponticum var, had showed the celluloytic activity. Chun (1977) also suggested that Trichoporon pullulons 225 isolated from the saline water of the Yeoung San River had a similar properties. However, the assay conditions for enzyme activity were not yet investigated. Thus, the present work was undertaken to examine some conditions for CMCase activity and at the same time to compare the activities of crude enzyme produce from above two species of Trichosporon pullulans. The results are as follows; 1. The maximum production of total reducing sugar by crude enzyme of Tr. pululans was after 30 minutes, whereas that of Tr. cutanuem FRI-425 was after 90 minutes. This fact showed that the reaction velocity of enzyme from Tr. pullulans 225 was more faster than that of Tr. cutaneum FRI-425. 2. Two species showed a similar trend to increase the production of reducing sugar in proportion to the increment in substrate concentration and to arrive at maximum level at lmg/ml of substrate concentration. However, Tr. pullulans 225 produced more $50{\mu}g$ of reducing sugar compared to Tr. cutaneum. 3. The optimum PH for CMCase activity is 5.0 for Tr. pullulans 225 as well as Tr. cutaneum FRI-425, and PH stability lie within the range of 6 and 8. In the activity and stability of enzyme on PH changes, enzyme of Tr. cutaneum FRI-425 was more unstable than that of TY. pullulans 225. 4. The optimum temperature for CMCase activity was $40^{\circ}C$, and enzyme activity from Tr. pullulans 225 was more sensitive to temperature changes compared with that of TY. cutaneum. The heat stability was within $40^{\circ}C$, but that was rapidly decreased above $40^{\circ}C$. In comparison of the heat stability for enzyme of Tr. cutaneum FRI-425 with that of Tr. pullulans 225 at the same temperature of $80^{\circ}C$, the former was some 10 percent more stable than the latter.

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Activity and Stability of Alcohol Oxidase from Hansenula sp. by Electrostimulation (전기자극에 의한 알콜 산화효소의 활성도와 안정도연구)

  • Lee, Kang-Min;Kim, Kyung-Suk;Park, Chung-Ung
    • Journal of the Korean Chemical Society
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    • v.48 no.2
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    • pp.171-176
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    • 2004
  • We investigated the activity and stability of alcohol oxidase from Hansenula sp. under the electric stimulation. The activity and stability of alcohol oxidase depended on electric output voltage, stimulation time, pulse duration and pulse interval. This inactivation of the enzyme under electric stimulation could be recovered by stabilizing additives such as sugars, polymers and hydrogels. The enzyme activity retained about 52% in 10% trehalose solution under electric stimulation with 40 V and 10 min. The stabilizing of enzymes against electric stimulation showed a great potential use of enzymes in biotechnology and medical engineering fields.

Conformational Switch and Functional Regulation of Proteins (단백질의 구조 전환과 기능 조절)

  • Yu, Myeong-Hee
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2001.11b
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    • pp.3-6
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    • 2001
  • In common globular proteins, the native form is in its most stable state. However, the native form of inhibitory serpins (serine protease inhibitors) and some viral membrane fusion proteins is in a metastable state. Metastability in these proteins is critical to their biological functions. Our previous studies revealed that unusual interactions, such as side-chain overpacking, buried polar groups, surface hydrophobic pockets, and internal cavities are the structural basis of the native metastability. To understand the mechanism by which these structural defects regulate protein functions, cavity-filling mutations of a 1-antitrypsin, a prototype serpin, were characterized. Increasing conformational stability is correlated with decreasing inhibitory activity. Moreover, the activity loss appears to correlate with the decrease in the rate of the conformational switch during complex formation with a target protease. We also increased the stability of a 1-antitrypsin greatly via combining various stabilizing single amino acid substitutions that were distributed throughout the molecule. The results showed that a substantial increase of stability, over 13 kcal/mol, affected the inhibitory activity with a correlation of 11% activity loss per kcal/mol. The results strongly suggest that the native metastability of proteins is indeed a structural design that regulates protein functions and that the native strain of a 1-antitrypsin distributed throughout the molecule regulates the inhibitory function in a concerted manner.

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A Study on the Image of Male Fashion Considering the Necktie Width and Body Type (넥타이의 폭과 체형을 고려한 남성패션 이미지 연구)

  • Choi, Su-Gyeong;Jeong, Su-Jin
    • Journal of the Korea Fashion and Costume Design Association
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    • v.11 no.1
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    • pp.111-121
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    • 2009
  • The purpose of this study is to investigate the effect of body type(thin, standard, fat), necktie width(narrow, medium, wide), gender(man, woman) on image formation. Sets of stimulus and response scales(7 point semantic) were used as experimental materials. The stimuli were 9 color pictures manipulated with the combination of necktie-width and body type using computer simulation. The subjects were 116 male undergraduates and 99 female undergraduates living in Gyeongsangnam-do. Image factor of the stimulus was composed of 4 different components, young-activity, attractiveness-gracefulness, stability, and boldness. In the young-activity, attractiveness-gracefulness and stability, body type and necktie-width showed independent effect. In the boldness, body type showed independent effect. Significant interaction effects of body type and necktie-width on young-activity, attractiveness-gracefulness and stability were found. For young-activity image, necktie of narrow width were effective. For attractiveness-gracefulness image, body type of standard were effective. For stability image, body type of standard and body type of thin on necktie of narrow and medium width were effective. The male fashion image can be perceived differently according to combination of the clues used in body type and necktie-width. Therefore, when the male fashion to create images that will have to consider the body type and necktie width.

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