• Title/Summary/Keyword: Leucine

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Studies on the Production of Serratiopeptidase from Serratia Culture (세라티아 배양에 의한 세라티오펩티다아제의 생산에 관한 연구)

  • 노현수;박호진;이병룡
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.207-212
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    • 1992
  • An anti-inflammatory agent, serratiopeptidase, was produced from the culture of the Serratia marcescens. The effects of carbon sources, nitrogen sources and inducers on the production were investigated. Citrate was found to be inhibitory in the production of serratiopeptidase. The enzyme was synthesized in the synthetic medium without inducers, albeit low level of synthesis. But the synthesis was increased by the addition of proteinaceous substrate and leucine. Induction of extracellular proteinase by its end-product was discovered, which is not common in the proteinase synthesis in the bacteria. By the glucose fed-batch culture, we found the possible catabolite repression on the production of serratiopeptidase.

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Phenylalanyl-2-Sulfanilylglycine as Substrate for Leucine Aminopeptidase Assay

  • Hwang, Se-Young;Cho, Suk-Young;Yoo, Ick-Dong
    • Journal of Microbiology and Biotechnology
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    • v.5 no.6
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    • pp.319-323
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    • 1995
  • A chromogenic mimic of phenlyalanyl-dipeptide, L-phenylalanyl-L-2-sulfanilylglycine (PSG), was synthesized and examined for its usability in leucine aminopeptidase (LAP) assay. The enzyme activity was easily determined by measuring the amount of diazotized adduct of sulfanilic acid released upon hydrolysis of PSG ($\varepsilon^{420}$=18,000/M/cm). Under the experimental conditions employed, PSG showed a Km of 0.063 mM and a Kcat of 1683/min, assessable less than 0.1 $\mu$ g of LAP per milliliter. And the presence of aminopeptidase M (APM) was suggested to be negligible in LAP assay. This novel assay can circumvent the occasional yellow background in biological systems, i.e., serums, etc..

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Comparison of Methods for the Measurement of Bacterial Production in a Lake Ecosystem (호수 생태계에서 세균 생산량 측정 방법의 비교)

  • 김명운;강찬수;김상종
    • Korean Journal of Microbiology
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    • v.28 no.4
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    • pp.318-323
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    • 1990
  • The bacterial secondary production was measured at 6 sites of Lake Soyang in October, 1989 by $^{3}$H-thymidine incorporation rate. Verfication for the method of bacterial secondary production measurement showed that $^{3}$H-thymidine incorporated into DNA, RNA and protein by average percentage of 38.45, 42.27 and 20.07%, respectively. THe more increased incoporated $^{3}$H-thymidine, the more increasde DNA fraction, but protein fraction was generally low. Incorporation of rate of /usp 3/H-thymidine. $^{3}$H-leucine into protein correlated with protein fraction of incorporated $^{3}$H-thymidine. Conversion factors were calculated as follows; $1.83*10 ^{20}$ cells/moles of thymidine incorporated/hr and 1.69*10$^{22}$ cells/moles of leucine incorporated/hr.

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Microwave Assisted Rapid Synthesis of Novel Optically Active Poly(amide-imide)s Based on N-Trimellitylimido-L-Leucine Diacid Chloride and Hydantoin Derivatives

  • Faghihi, Khalil
    • Macromolecular Research
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    • v.12 no.3
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    • pp.258-262
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    • 2004
  • We have developed facile and rapid polycondensation reactions of N-trimellitylimido-L-leucine diacid chloride 1 with eight different derivatives of hydantoin compounds 2a-h, in the presence of a small amount of a polar organic medium, such as Ο-cresol, by using a domestic microwave oven. The polycondensation reactions proceeded rapidly-they were complete within 7-9 min-to produce a series of novel optically active poly(amide-imide)s (3a-h) in high yield with inherent viscosities of 0.33-0.51 dL/g. We characterized the resulting poly(amide-imide)s by elemental analysis, thermal gravimetric analysis (DSC, TGA, and DTG), and FTIR spectroscopy, and by measuring their viscosities, specific rotations, and solubilities. All of the polymers were soluble at room temperature in polar solvents such as N ,N-dimethylacetamide, N,N-dimethylformamide, dimethylsulfoxide, tetrahydrofuran, and N-methyl-2-pyrrolidone.

Preparation of Glutamic Acid-Leucine Copolymer Containing Indomethacin for Controlled Delivery

  • Yeom, Young-Il;Kim, Hyun-Pyo;Kim, Hack-Joo;Byun, Si-Myung;Kim, Nam-Deuk
    • Bulletin of the Korean Chemical Society
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    • v.7 no.3
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    • pp.213-217
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    • 1986
  • A series of copolypeptides of glutamic acid and leucine have been synthesized by N-carboxy-${\alpha}$-amino acid anhydride procedure and cast to form injectable microparticulate monolithic devices in which indomethacin was physically dispersed. With these devices, various release properties and possible clinical application were studied. The release rate of the drug had a close relationship with the monomer composition of the copolymer matrix as well as the environmental pH condition. The monolithic device of glutamic acid/leucine = 50/50 was found to be the most promising one as a ploymeric delivery system of indomethacin. The intrinsic viscosity of this copolymer was 4.35 dl/g and the release rate was 18.5${\mu}g/g/day$.

Isolation and Characterization of a Basic Leucine Zipper Gene in Poplar (Populus alba × P. glandulosa) (현사시나무에서 Basic Leucine Zipper 유전자의 분리와 특성 구명)

  • Yoon, Seo-Kyung;Lee, Hyoshin;Bae, Eun-Kyung;Choi, Young-Im;Kim, Joon-Hyeok;Noh, Seol Ah
    • Journal of Korean Society of Forest Science
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    • v.103 no.2
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    • pp.189-195
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    • 2014
  • Basic leucine zipper (bZIP) protein is a regulatory transcription factor that plays crucial roles in growth, development and stress response of plant. In this study, we isolated a PagbZIP1 gene that belonged to Group SE3 of bZIP from Populus alba ${\times}$ P. glandulosa, and investigated its expressional characteristics. The PagbZIP1 is 844 base pairs long and encodes a putative 144-amino-acid protein with an expected molecular mass of 16.6 kDa. The PagbZIP1 has two conserved domains including the basic and leucine zipper portions. Southern blot analysis revealed that two copies of the gene are presented in the poplar genome. PagbZIP1 was specifically expressed in the root and suspension cells. Moreover, the expression of PagbZIP1 was induced by drought, salt, cold and ABA. Therefore, our results indicated that PagbZIP1 might be expressed in response to abiotic stress through the ABA-mediated signaling pathway in poplar.

Enhanced Permeation of Leucine Enkephalin and [D-Ala2]-leucine Enkephalinamide across Nasal, Rectal and Vaginal Mucosae of Rabbit (토끼의 비강, 직장 및 질 점막을 통한 로이신엔케팔린과 [D-알라2]-로이신엔케팔린아미드의 투과 증진)

  • 전인구;박인숙;곽혜선
    • Biomolecules & Therapeutics
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    • v.10 no.2
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    • pp.104-113
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    • 2002
  • The effects of enzyme inhibitors and penetration enhancers on the permeation of leucine enkephalin (Leu-Enk) and its synthetic analog, [${D-ala}^2$]-leucine enkephalinamide (YAGFL) across the nasal, rectal and vaginal mucosae were evaluated. Enzyme inhibitors and penetration enhancers employed for Leu-Enk permeation study were amastatin(AM), thimerosal(TM) and ethylenediaminetetraacetic acid disodium salt(EDTA), and sodium taurodihydrofusidate (STDHF). Those for YAGFL permeation study were TM, benzalkonium chloride(BC) and EDTA, and STDHF, sodium deoxycholate(SDC), sodium glycholate(SGC), glycyrrhizic acid ammonium salt (GAA), L-$\alpha$-Iysophosphatidylcholine(LPC) and mixed micelle (MM, STDHF: linoleic acid = 15 mM : 5 mM). The addition of TM alone on the donor and receptor solutions for Leu-Enk permeation study across all the three kinds of mucosae failed to inhibit the degradation; it completely degraded in 6 hrs, and no permeation occurred. However, with addition of three kinds of inhibitors together, the fluxes across nasal, rectal and vaginal mucosae were $\20.7{pm}2.5$>/TEX>,$\0.3{pm}0.05$>/TEX> and $\1.4{pm}0.5$ $\mu$\mid$textrm{m}$/$\textrm{cm}^2$/hr, respectively. Moreover, the addition of STDHF in the presence of the above three inhibitors enhanced permeation across nasal, rectal and vaginal mucosae 1.3, 15 and 1.3 times, respectively. YhGFL also degraded in the donor and receptor solutions rapidly as time went. With mixed inhibitors of TM and EDTA, the percents of YAGFL remaining in the donor solutions facing nasal, rectal and vaginal mucosae were 69.7, 69.8 and 79.8%, respectively; the percent permeated increased to 10, 2.1 and 5.7%, respectively. The addition of STDHF in the presence of either BC/EDTA or TW/EDTA increased the permeation 2.2, 11.0 and 2.9 times, and 2.21, 14.0 and 2.7 times for nasal, rectal and vaginal mucosae, respectively. With SDC, SGC, GAA, LPC ud MM in the presence of TM/EDTA increased permeation; especially, they increased permeation across vaginal mucosae effectively, and the enhancement factors were 12.5, 7.6, 8.7, 5.7 and 5.5, respectively. The degradation extent of YAGFL was correlated with protein concentrations in the epidermal and serosal extracts. The flux of YAGFL across nasal mucosa increased dose-dependently.

Changes of Chemical, Bacteriological, and Allergenicity of Raw Milk by Gamma Irradiation (감마선 조사 처리에 의한 우유의 화학적${\cdot}$세균학적 및 항원성 변화)

  • Noh, Yeong-Bae;Kim, Seung-Il;Kim, Hyeon-Su;Jeong, Seok-Geun;Chae, Hyeon-Seok;An, Jong-Nam;Jo, Cheol-Hun;Lee, Wan-Gyu;Ham, Jun-Sang
    • Journal of Dairy Science and Biotechnology
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    • v.23 no.2
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    • pp.93-98
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    • 2005
  • Effects of heat and gamma irradiation on chemical, microbiological, and immunological changes of raw milk were compared. Free fatty acid content of milk showed increasing tendency according to the increase of heating temperature and irradiation dose, and showed similarity in UHT (ultra high temperature) and 5 kGy irradiation. Total bacterial counts and coliforms were not detected after treatment of LTLT (low temperature long time), HTST (high temperature short time), UHT, and irradiation from 1 to 10 kGy in the milk with initial microbial load at $10^3$ CFU/mL initially, but after 7 day storage, were not detected in UHT milk and that irradiated at 3 kGy or above. Heat treatment decreased (p<0.05) arginine, asparate, iso-leucine, lysine, and methionine content compared to raw milk while irradiation decreased (p<0.05) asparate, histidine, iso-luecine, leucine, and lysine content, which means irradiation could change primary structure of milk proteins. It was concluded that f kGy gamma irradiation treatment of raw milk could give a similar effect to UHT treatment in chemical and microbiological viewpoint, and may reduce allergenicity of raw milk.

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Inhibition of Enzymatic Degradation of Leucine Enkephalin and $[D-Ala^2]$-Leucine Enkephalinamide in Various Rabbit Mucosal Extracts by Inhibitors (효소 억제제에 의한 토끼의 점막 추출액중 로이신엔케팔린 및 [D-알라$^2$-로이신엔케팔린아미드의 분해 억제)

  • Chun, In-Koo;Park, In-Sook;Hyun, Jeen
    • Journal of Pharmaceutical Investigation
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    • v.26 no.3
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    • pp.175-185
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    • 1996
  • To inhibit the enzymatic degradation of leucine enkephalin (Leu-Enk) and its synthetic analog. $[D-ala^2]$-leucine enkephalinamide (YAGFL), in the nasal, rectal and vaginal mucosal and serosal extracts of rabbits, effects of enzyme inhibitors such as amastatin (AM), puromycin (PM), thiorphan (TP), thimerosal (TM), EDTA, N-carboxymethyl-Phe-Leu (CPL), phenylethyl alcohol (PEA), phenylmercuric acetate (PMA), benzalkonium chloride (BC) and modified cyclodextrins, alone or in combination, were observed by assaying the pentapeptides staying intact during incubation. Mucosa extracts were prepared by exposing freshly-excised mucosal specimens mounted on Valia-Chien cells to isotonic phosphate buffer while stirring. The degradation of Leu-Enk and YAGFL followed the apparent first-order kinetics. The half-lives (mean) in the nasal, rectal and vaginal mucosal extracts were found to be 1.07, 0.33 and 1.14 hr for Leu-Enk, and 16.9, 6.2 and 6.8 hr for YAGFL, respectively. AM or PM, which is an aminopeptidase inhibitor, did not show a sufficient inhibition of Leu-Enk $(50\;{\mu}g/ml)$ degradation in all kinds of extracts. $Dimethyl-{\beta}-cyclodextrin\;(DM-{\beta}-CyD)$ decreased the degradation rate constants of Leu-Enk about 2 or 3 times, comparing with no additive. However, the use of mixed inhibitors of AM $(50\;{\mu}M)$/TM (0.25 mM)/EDTA (5 mM) resulted in a full stabilization of Leu-Enk by decreasing the degradation rate constants 67.3, 161.3 and 113.8 times far the nasal, rectal and vaginal mucosal extracts, respectively, comparing with no inhibitor. With mixed inhibitors, Leu-Enk remained intact more than 90% after 6 hr-incubation. In the stabilization of YAGFL, hM, TP or CPL alone showed little efffct, and some additives demonstrated a considerable inhibition of YAGFL degradation in the rank order of TM > BC > EDTA. However, the addition of mixed inhibitors such as TM (0.5 mM) and EDTA (5 mM) into the extracts protected YAGFL from the degradation by more than 85% even after 24 hr-incubation, suggesting almost complete inhibition of YAGFL degradation in the extract. On the other hand, $DM-{\beta}-CyD\;or\;hydroxypropyl-{\beta}-cyclodextrin$ (10%) were also found to retard enzymatic degradation rates of YAGFL markedly, and resulted in staying intact more than 80% of YAGFL in the nasal and vaginal mucosal extracts, and more than 60% in the rectal mucosal extract after 16 hr-incubation.

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A Trial for Utilizing Flounder Skin Gelatin as an Emulsifier through Enzymatic Modification (가자미피 젤라틴의 효소적 수식에 의한 유화제의 시제)

  • KIM Se-Kwon;JEON You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.5
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    • pp.345-355
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    • 1991
  • In order to effectively utilize the by-products of sea-food, the utilization of enzyme-modified flounder(Limanda aspera) skin gelatin as an emulsifier was investigated. In the experiment, the gelatin was extracted from the flounder skin with the heat-treatment at $60^{\circ}C$ and in pH 5.0 for 3 hrs with four volumes of distilled water and emulsifiers were enzymatically modified L-leucine alkyl esters$(L-leucine-OC_n$ : n= 2, 4, 6, 8 and 10) to the gelatin$(EMFSG-C_2,\;EMFSG-C_4,\;EMFSG-C_6,\;EMFSG-C_8,\;EMFSG-C_{10})$ for improving the functional properties such as emulsifying activity, emulsifying viscosity, whippability, electric conductivity, critical micelle concentration and interface tension, etc. Also, the functional properties of the L-leucine alkyl ester modified gelatins were compared with those of Tween-60 as reference. Molecular weights of the enzymatically modified flounder skin gelatin(EMFSG) were 20.5kDa. in $EMFSG-C_2.\;19.5 kDa.\;in\;EMFSG-C_4\;and\;16.5kDa.\;in\;EMFSG-C_6,\;EMFSG-C_8$ and $EMFSG-C_{10}$. respectively. Emulsifying activity and emulsifying viscosity in the modified gelatins were risen with increase of carbon number of the introduced L-leucine alkyl esters. Among the modified gelatins, $EMFSG-C_6$ exhibited the highest emulsifying stability and foaming stability, whereas $EMFSG-C_8$ showed the highest whippability. The electric conductivities of the all $EMFSG-C_n$ were linearly risen to critical micelle concentration(CMC) , therefore $EMFSG-C_{10}$ exhibited the lowest CMC value and interface tension, and dense particles in the microscopic observation. In conclusion, the best quality in functional properties was assured on $EMFSG-C_{10}$.

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