• 제목/요약/키워드: enzymatic degradation

검색결과 197건 처리시간 0.028초

황체호르몬 유리호르몬(LHRH)의 경점막 수송: 토끼 점막균질액 중에서 $[D-Ala^6]$ LHRH의 효소적 분해 특성 및 중쇄지방산염의 안정화 효과 (Transmucosal Delivery of Luteinizing Hormone-Releasing Hormone(LHRH): Enzymatic Proteolysis of $[D-Ala^6]$ LHRH and Inhibitory Effect of Medium Chain Fatty Acid Salts in Rabbit Mucosa)

  • 박정숙;정연복;한건
    • 약학회지
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    • 제38권2호
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    • pp.202-210
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    • 1994
  • To investigate the feasibility of mucosal delivery of $[D-Ala^6]$ LHRH, a potent analogue of LHRH, enzymatic proteolysis of $[D-Ala^6]$ LHRH and inhibitory effect of medium chain fatty acid salts(MFA) were studied using rabbit mucosal homogenate. $[D-Ala^6]$ LHRH incubated in homogenates of rectal(RE), nasal(NA) and vaginal(VA) mucosa were assayed by HPLC. The degradation of $[D-Ala^6]$ LHRH followed the first order kinetics. The degradation products were found as $[D-Ala^6]$ $LHRH^{1-7}$(m-i), to a lesser extent, $[D-Ala^6]$ $LHRH^{1-9}$(m-ii) and $[D-Ala^6]$ $LHRH^{1-3}$(m-iii) by the method of amino acid analysis(PITC method). The formation of$[D-Ala^6]$ $LHRH^{1-7}$ was not inhibited by the addition of disodium ethylenediaminetetraacetic acid but inhibited by sodium tauro-24,25-dihydrofusidate, suggesting that endopeptidase 24.11(EP 24.11) cleaves the $Leu^7-Arg^8$ bond of $[D-Ala^6]$ LHRH and is the primary $[D-Ala^6]$ LHRH degrading enzyme. The patterns of $[D-Ala^6]$ LHRH degradation indicated that EP 24.11 exists in each mucosal homogenate with the order of RE>NA>VA. MFA significantly inhibited the proteolysis of $[D-Ala^6]$ LHRH. The addition of sodium caprate(1.0%) or sodium laurate(0.5%) to the each mucosal homogenate completely protected $[D-Ala^6]$ LHRH from the degradation.

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Degradation of Clavulanic Acid During the Cultivation of Streptomyces clavuligerus; Instability of Clavulanic Acid by Metabolites and Proteins from the Strain

  • Ishida Kenji;Hung Trinh Viet;Lee Hei-Chan;Liou Kwang-Kyoung;Shin Chang-Hun;Yoon Yeo-Joon;Sohng Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • 제16권4호
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    • pp.590-596
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    • 2006
  • Clavulanic acid (CA) produced by Streptomyces clavuligerus is degraded during the bacterial cultivation. The degradation was examined in three different aspects, including physical, chemical, and enzymatic effects, in order to understand the degradation during the cultivation. The result showed that CA was unstable in the production medium containing ammonium salts and amino acids, owing to ammonium ions and amine groups. In addition, the degradation was not only due to instability of CA by metabolites and proteins, but also enzymes from S. clavuligerus such as $\beta-lactamase$ and penicillin-binding proteins. However, the degradation caused by these enzymes was not highly significant compared with the degradation during the cultivation, owing to irreversible reactions between CA and enzymes.

토끼의 수종 점막 추출액중 $[D-Ala^2]-Methionine$ Enkephalinamide의 분해 및 안정화 (Degradation and Stabilization of $[D-Ala^2]-Methionine$ Enkephalinamide in Various Rabbit Mucosa Extracts)

  • 전인구;양윤정
    • Journal of Pharmaceutical Investigation
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    • 제22권3호
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    • pp.173-183
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    • 1992
  • To study the feasibility of transmucosal delivery of $[D-ala^2]-methionine$ enkephalinamide (YAGFM), its enzymatic degradation and stabilization in various rabbit mucosal extracts were investigated by HPLC method. The degradation of YAGFM was observed to follow the first-order kinetics and the half-lives of YAGFM in the nasal, rectal and vaginal mucosal extracts were found to be 25.7, 3.0 and 7.8 hr, respectively. However, there was no significant difference in degradation rates of YAGFM between the mucosal and serosal extracts obtained from the same mucosal membrane. This finding suggests that even a synthetic enkephalin analog, which is designed to be resistent to aminopeptidases, needs to be fully protected from the enzymatic degradation in mucosal sites for the delivery of the analog through mucosal routes. To inhibit the degradation of YAGFM in various mucosal extracts, effects of enzyme inhibitors such as bestatin (BS), amastatin (AM), thiorphan (TP), thimerosal (TM) and EDTA, alone or in combination, and modified cyclodextrins were observed by assaying YAGFM staying intact during 24 hr-incubation at $37^{\circ}C$. It was found from the results that mixed inhibitors such as TM (0.5 mM)/EDTA (5 mM) or AM $(50{\mu}M)/TM$ (0.5 mM)/EDTA (5 mM) provided very useful means for the stabilization in various mucosal extracts. The latter was found to protect YAGFM from the degradation in the nasal, rectal, and vaginal mucosal extracts by 90.9, 90.4 and 91.3%, respectively, after 24 hr-incubation, suggesting almost complete inhibition of YAGFM-degrading enzymes present in the incubation mixture. However, BS $(50{\mu}M)$, AM 50 $(50{\mu}M)$ or TP$(50{\mu}M)$ alone did not reveal sufficient inhibition except TM (0.5 mM) or EDTA (5 mM). The adddition of $2-hydroxylpropyl-{\beta}-cyclodextrin$(10%) to the nasal mucosal extract, and $dimethyl-{\beta}-cyclodextrin$(10%) to the rectal and vaginal mucosal extracts reduced the first-order rate constants for the degradation of YAGFM by 5.8, 17.3 and 8.9 times, respectively, compared to those with no additive.

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순천만 갯벌 토양의 섬유소 분해능 및 체외효소 활성 (Cellulose Degradation and Extracellulat Enzymatic Activity of the Mud Flat in Sunchon Bay)

  • 백근식;최지혁;성치남
    • 미생물학회지
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    • 제36권2호
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    • pp.130-135
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    • 2000
  • 순천만 갯벌의 3개 지점의 표층(5cm)과 심층(20 cm)토양의 물리화학적 요인과 종속영양세균의 분포, 섬유소 분해율과 체외효소의 활성을 측정하였다. 온도, 수분함량, 인산염 인 그리고 유기물함량은 각각 -1~$30^{\circ}C$, 42.1~53.1%, 0.0779~0.1961mg/g, 그리고 1.99~7.64%로 나타나었다. 섬유소 films의 분해율은 7.7%~100%/month 범위로 1~2월에 최저 그리고 8~9월에 최고의 분해율을 나타냈다. 종속영양세균의 분포는 $0.87{\times}10^6 ~ 3.6{\times}10^7 $CUFs/g dry soil의 범위에 속해Tdmu 심층의 경우 표층에 비해 낮았다. MUF-기질의 분해율로 측정한 phoshatase, $\alpha$-D-gluocosidase, $\beta$-D-glucosidase, cellobiohydrolase 활성의 변화는 각각 152.23~1779.80 nMi/hr 2.67~202.18 nM/hr, 5.03~258.26 nM/hr, 3.42~63.07 nM/hr으로 모두 하계에 상대적으로 높은 값을 나타내었다. 섬유소 분해는 수온상승이 주된 요인이었으며, 정점간, 깊이간 큰 차이는 나타나지 않았다. 체외효소의 활성은 온도 및 섬유소 분해율과 높은 양의 상관관계를 나타냈다. 종속영양세균의 분포는 섬유소 분해와 체외효소의 활성과 높은 상괸은 나타나지 않았다.

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Cross-Linked Starch Microspheres: Effect of Cross-Linking Condition on the Microsphere Characteristics

  • Atyabi, Fatemeh;Manoochehri, Saeed;Moghadam, Shadi H.;Dinarvand, Rassoul
    • Archives of Pharmacal Research
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    • 제29권12호
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    • pp.1179-1186
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    • 2006
  • Cross-linked starch microspheres were prepared using different kinds of cross-linking agents. The influence of several parameters on morphology, size, swelling ratio and drug release rate from these microspheres were evaluated. These parameters included cross-linker type, concentration and the duration of cross-linking reaction. Microspheres cross-linked with glutaraldehyde had smooth surface compared with those prepared with epichlorhydrine or formaldehyde. The particle size increased with increasing the cross-linking time and increasing the drug loading. Swelling ratio of the particles was a function of cross-linker type but not the concentration or time of cross-linking. Drug release from starch microspheres was measured in phosphate buffer and also in phosphate buffer containing a-amylase. Results showed that microspheres cross-linked with epichlorhydrine released all their drug content in the first 30 minutes. However, cross-linking of the starch microspheres with glutaraldehyde or formaldehyde decreased drug release rate. SEM and drug release studies showed that cross-linked starch microspheres were susceptible to the enzymatic degradation under the influence of alpha-amylase. Changing the enzyme concentration from 5000 to 10,000 IU/L, increased drug release rate but higher concentration of enzyme (20,000 IU/L) caused no more acceleration.

리그닌/PVA 나노섬유 웹의 수분 특성 및 생분해성 평가 (Water Absorption Properties and Biodegradability of Lignin/PVA Nanofibrous Webs)

  • 송유정;이은실;이승신
    • 한국의류학회지
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    • 제41권3호
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    • pp.517-526
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    • 2017
  • The biodegradation and water absorption properties of lignin/poly(vinyl alcohol) (PVA) nanofibrous webs are investigated. Lignin/PVA nanofibrous webs containing 0, 50, and 85wt% of lignin were prepared via an electrospinning process to observe the effect of the lignin concentration on the biodegradability and water absorption properties of lignin/PVA nanofibrous webs. The morphology of the materials was examined by field emission scanning electron microscopy (FE-SEM) and atomic force microscopy (AFM). To understand the wetting behavior and hydrophilic nature of the electrospun lignin/PVA nanofibrous webs, the water absorbency, contact angle, and water uptake were examined. The enzymatic degradation of lignin/PVA nanofibrous webs was investigated using laccase by measuring total organic carbon (TOC) concentration over a course of 50 days. Water drops were absorbed immediately into all of the specimens. The water uptake of lignin/PVA nanofibrous webs increased as the amount of PVA in the lignin/PVA hybrid webs increased. The enzymatic degradation experiment indicated that the inherent biodegradability of lignin was retained after its transformation into nanofibers. Our findings imply that blending these two types of polymers is promising because it can lead to the development of a new range of multifunctional materials such as antimicrobial absorbent nanotextiles based on sustainable biopolymers.

Bacillus licheniformis 단백질 분해 효소에 의한 정연 견사의 특성 I. 정연 견사의 이화학적 특성 (Physicochemical Characteristics of Silk Fibroin Degummed by Protease in Bacillus licheniformis I. Physicochemical Characteristics of Degummed Silk Fiber)

  • 김영대;남중희
    • 한국잠사곤충학회지
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    • 제34권2호
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    • pp.41-51
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    • 1992
  • 견사의 정연 방법으로는 알카리 용액에서 열을 이용하는 방법과 단백질 분해 효소를 이용하는 방법으로 크게 나눌 수 있는데 본 실험은 Bacillus licheniformis 단백질 분해 효소로 정연한 견사의 이화학적 특성과 열 처리 영향 및 효소 정연 견 피브로인 수용액의 특성을 비누 정연 견사와 비교하여 실험한 결과 다음과 같은 결과를 얻었다. 1. 정연 견사의 표면을 주사 전자현미경으로 관찰한 결과 정연 방법에 따른 뚜렷한 차이가 없었으며 정연 견사의 아미노산 조성을 분석한 결과에서도 정연 방법에 따른 큰 차이가 없었다. 2. 효소 정연 견사는 비누 정연 견사에 비하여 강력과 신도가 약간 저하되었고 점도도 비누 정연 견사의 0.59에 비하여 효소 정연 견사는 0.56으로서 저하된 반면 ($\alpha$$\varepsilon$) amino group content는 비누 정연견사의 49.5$\mu$mo1/g보다 효소 정연 견사는 53.6 $\mu$mol/g로서 증가되었으므로 효소 정연 견사는 비누 정연 견사보다 degradation이 더 일어난 것으로 추측된다. 3. 효소 정연 견사는 비누 정연 견사에 비하여 복굴절도가 높았으며 DSC thermogram에서 열 분해 온도도 약 1$0^{\circ}C$ 높았다. 또 효소 정연 견사의 IR 및 X-ray crystallinity도 각각 0.70과 0.53으로서 비누정연 견사의 0.67과 0.50에 비하여 약간 높았으므로 효소 정연 견사의 경우 결정성 및 배향성이 비누 정연 견사보다 높은 것으로 판단된다.

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Enzymatic and Non-enzymatic Degradation of Poly (3-Hydroxybutyrate-co-3-Hydroxyvalerate) Copolyesters Produced by Alcaligenes sp. MT-16

  • Choi Gang Guk;Kim Hyung Woo;Rhee Young Ha
    • Journal of Microbiology
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    • 제42권4호
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    • pp.346-352
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    • 2004
  • Poly(3-Hydroxybutyrate-co­3-Hydroxyvalerate), poly(3HB-co-3HV), copolyesters with a variety of 3HV contents (ranging from 17 to $60\;mol\%$) were produced by Alcaligenes sp. MT-16 grown on a medium containing glucose and levulinic acid in various ratios, and the effects of hydrophilicity and crystallinity on the degradability of the copolyesters were evaluated. Measurements of thermo-mechanical pro­perties and Fourier-transform infrared spectroscopy in the attenuated total reflectance revealed that the hydrophilicity and crystallinity of poly(3HB-co-3HV) copolyesters decreased as 3HV content in the copolyester increased. When the prepared copolyester film samples were non-enzymatically hydrolysed in 0.01 N NaOH solution, the weights of all samples were found to have undergone no changes over a period of 20 weeks. In contrast, the copolyester film samples were degraded by the action of extra­cellular polyhydroxybutyrate depolymerase from Emericellopsis minima W2. The overall rate of weight loss was higher in the films containing higher amounts of 3HV, suggesting that the enzymatic degra­dation of the copolyester is more dependent on the crystallinity of the copolyester than on its hydro­philicity. Our results suggest that the degradability characteristics of poly(3HB-co-3HV) copolyesters, as well as their thermo-mechanical properties, are greatly influenced by the 3HV content in the copoly­esters.

Allergenic potential and enzymatic resistance of buckwheat

  • Lee, Sujin;Han, Youngshin;Do, Jeong-Ryong;Oh, Sangsuk
    • Nutrition Research and Practice
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    • 제7권1호
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    • pp.3-8
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    • 2013
  • Buckwheat is known as a health food but is one of the major food allergens triggering potentially fatal anaphylaxis in Asia, especially in Japan and Korea. This study was conducted to investigate the characteristic of enzymatic resistance of buckwheat protein and allergenic potential. Enzymatic resistance of buckwheat protein was performed with in vitro digestibility test in simulated gastric fluid (SGF), pH 1.2, using pepsin and simulated intestinal fluid (SIF) using chymotrypsin. Reactivity of buckwheat proteins to human IgE was performed using six allergic patients sensitized to buckwheat. Buckwheat's IgE levels were measured using the Phadia UniCAP-system. Buckwheat protein, 16 kDa, still remained after 30 min treatment of pepsin on SDS-PAGE. Even though 16 kDa almost disappeared after 60 min treatment, two out of the six buckwheat patients' sera showed reactivity to hydrolysate after 60 min treatment, indicating that allergenicity still remained. In simulated intestinal fluid (SIF) using chymotrypsin, buckwheat protein, 24 kDa, showed resistance to hydrolysis with chymotrypsin on SDS-PAGE, and still had allergenicity based on the result of ELISA. Our results suggest that buckwheat proteins have strong resistance to enzyme degradation. This may be attributed in part to the allergenic potential of buckwheat. Further study should be continued regarding buckwheat allergy.