• 제목/요약/키워드: 5-Amino-3H-1

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NADP+-Dependent Dehydrogenase SCO3486 and Cycloisomerase SCO3480: Key Enzymes for 3,6-Anhydro-ʟ-Galactose Catabolism in Streptomyces coelicolor A3(2)

  • Tsevelkhorloo, Maral;Kim, Sang Hoon;Kang, Dae-Kyung;Lee, Chang-Ro;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제31권5호
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    • pp.756-763
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    • 2021
  • Agarose is a linear polysaccharide composed of ᴅ-galactose and 3,6-anhydro-ʟ-galactose (AHG). It is a major component of the red algal cell wall and is gaining attention as an abundant marine biomass. However, the inability to ferment AHG is considered an obstacle in the large-scale use of agarose and could be addressed by understanding AHG catabolism in agarolytic microorganisms. Since AHG catabolism was uniquely confirmed in Vibrio sp. EJY3, a gram-negative marine bacterial species, we investigated AHG metabolism in Streptomyces coelicolor A3(2), an agarolytic gram-positive soil bacterium. Based on genomic data, the SCO3486 protein (492 amino acids) and the SCO3480 protein (361 amino acids) of S. coelicolor A3(2) showed identity with H2IFE7.1 (40% identity) encoding AHG dehydrogenase and H2IFX0.1 (42% identity) encoding 3,6-anhydro-ʟ-galactonate cycloisomerase, respectively, which are involved in the initial catabolism of AHG in Vibrio sp. EJY3. Thin layer chromatography and mass spectrometry of the bioconversion products catalyzed by recombinant SCO3486 and SCO3480 proteins, revealed that SCO3486 is an AHG dehydrogenase that oxidizes AHG to 3,6-anhydro-ʟ-galactonate, and SCO3480 is a 3,6-anhydro-ʟ-galactonate cycloisomerase that converts 3,6-anhydro-ʟ-galactonate to 2-keto-3-deoxygalactonate. SCO3486 showed maximum activity at pH 6.0 at 50℃, increased activity in the presence of iron ions, and activity against various aldehyde substrates, which is quite distinct from AHG-specific H2IFE7.1 in Vibrio sp. EJY3. Therefore, the catabolic pathway of AHG seems to be similar in most agar-degrading microorganisms, but the enzymes involved appear to be very diverse.

Arthrobacter luteus로부터 유래한 염기성 AL-Protease의 효소학적 성질 및 활성 아미노산 잔기의 검색 (Enzymological Properties of the Alkaline AL-Protease from Arthrobacter luteus and Detection of Its Active Amino Acid Residue)

  • 오홍록;상원태생;반진승
    • 한국식품영양과학회지
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    • 제13권2호
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    • pp.193-204
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    • 1984
  • Zymolyase 조효소로부터 분리, 정제되었고, 효모세포벽 용해 촉진물질로 밝혀진 바 있는 Arthrobacter luteus로부터 유래한 염기성 protease(AL-protease)의 효소학적 성질 및 활성 아미노산 잔기를 검색한 결과는 다음과 같다. 1. AL-pretense는 저해제 DFP 및 PMSF에 의해서 그 Protease 활성 및 용해 촉진활성이 동시에 완전히 소멸 되었으며, 그 저해 반응속도는 chymotrypsin에 대한 것에 비하여 대단히 완만하였다. 1.반응에서 AL-protease와 DFP의 결합 mole비는 1:1로 추정 되었다. 2. 정제된 AL-protease의 동결건조품 중에는 종래효모세포벽 용해반응에 관여하는 것으로 알려진 yeast phosphomannase를 비롯한 다당류 가수분해효소들의 활성은 그 어느 것도 인정되지 않았다. 3. AL-protease의 casein에 대한 최적 pH 및 최적 온도는 pH 10.5와 $65^{\circ}C$이었고, 그 활성은 pH 5${\sim}$11 사이와 $65^{\circ}C$이하에서 안정하였다. 또한, AL-Protease의 활성에 미치는 여러가지 금속이온의 영향은 인정되지 않았다. 4. [$^{32}P$]-DFP에 의하여 화학수식된 [$^{32}P$]-DIP-AL-protease에 대한 활성부위의 아미노산 잔기를 검색, 동정하기 위하여 조제용 PAG-전기영동, SDS-PAG-전기영동, Dowex 이온교환 크로마토그래피 및 고압 여지 전기영동을 실시하였고, 그 결과, AL-protease는 활성 부위에 1분자당 1 mole의 serine 잔기를 가지는 염기성 protease로 밝혀졌다.

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Regulation and Inactivation of Brain Phosphocholine-Phosphatase Activity

  • Seo, Seong-Kon;Liu, Xi-Wen;Lee, Hyun-Jeong;Kim, Hye-Kyeong;Kim, Mee-Ree;Sok, Dai-Eun
    • Archives of Pharmacal Research
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    • 제22권5호
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    • pp.464-473
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    • 1999
  • Regulation of phosphcholine-hydrolyzing phosphatase (phosphocholine-phosphatase) activity, purified from bovine brain, was examined under physiological conditions. Various endogenous phosphomonoesters, which were utilized as substrate, inhibited the phosphocoline-phosphatase activity competitively (Ki 5.5-$82.0 {\mu}M$); among phosphomonoesters tested, there was a similar order of capability between the binding affinity of substrate and the inhibitory potency. In addition, phosphate ions also inhibited the phosphatase activity competitively with a Ki value of approximately $16{\mu}M$. Although leucine or theophylline inhibited the phosphatase activity at pH 9.0, their inhibitory action decreased greatly at pH 7.4. The pH-Km and pH-Vm profiles indicate that ionizable amino acids are involved in substrate binding as well as catalysis, alluding that the phosphatase activity may be highly dependent on the intracellular pH. Amino acid modification study supports the existence of tyrosine, arginine or lysine residue in the active site, and the participation of tyrosine residue in the catalytic action may e suggested positively for the susceptibility to the action of tetranitromethane or HOl-generator. Separately, the oxidative inactivation of phosphocholine-phosphatase activity was investigated. Of oxidants tested, HOONO, HOCl, HOl and $ascorbate/Cu^{2+}$ system were effective to inactivate the phosphatase activity. Noteworthy, a remarkable inativation was accomplished by $30{\mu}M$ HOCl in combination with 1 mM Kl. Inaddition, $Cu^{2+}(3{\mu}M) $in combination with ascorbate at concentrations as low as 0.1-0.3 mM reduced the phosphatase activity to a great extent. From these results, it is proposed that the phosphocholine-phosphatase activity may be regulated endogenously and susceptible to the various oxidant system in vivo.

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Effects of alpha-linolenic acid and essential amino acids on the proliferation and differentiation of C2C12 myoblasts

  • Zhou, Dongjie;Li, Xiao-Han;Lee, Song‑Hee;Heo, Geun;Cui, Xiang-Shun
    • 한국동물생명공학회지
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    • 제37권1호
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    • pp.17-26
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    • 2022
  • Alpha-linolenic acid is an important polyunsaturated fatty acid that exhibits anticancer, anti-inflammatory, and antioxidative effects. In this study, we investigated the protective effects of alpha-linolenic acid on the cell proliferation and differentiation of C2C12 cells under essential amino acid-deficient conditions. Different concentrations of alpha-linolenic acid and essential amino acids were added to the growth and differentiation media. The concentrations of 10 µM of alpha-linolenic acid and 2% essential amino acid were chosen for subsequent experiments. Supplementation with alpha-linolenic acid and essential amino acids improved the proliferation and differentiation of C2C12 cells and significantly increased the mRNA levels of catalase, superoxide dismutase, B-cell lymphoma-2, and beclin-1 as well as the protein levels of PPARγ coactivator-1α compared to those in the controls. Moreover, supplementation with alpha-linolenic acid and essential amino acids reduced the levels of phosphorylated H2A.X variant histone, Bcl-2-associated X, p53, and light chain 3 during C2C12 cell proliferation, and increased the expression levels of myogenic factors 4 (myogenin) and 5 during C2C12 cell differentiation. Overall, we determined that alpha-linolenic acid and essential amino acids maintained the cell proliferation and differentiation of C2C12 cells via their anti-oxidative, anti-apoptotic, and anti-autophagic effects.

Calocybe indica의 배양적 특성과 균사 배양 적합 조건 설정 (Culture Characteristics and Optimal Conditions for Mycelial Growth of Calocybe indica)

  • 민경진;박혜성;이은지;이찬중
    • 한국균학회지
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    • 제48권3호
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    • pp.273-284
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    • 2020
  • 본 연구에서는C. indica 자실체의 안정적인 대량생산에 기반이 되는 필수적인 균사체 생산을 위한 최적 배양조건을 구명 하고자하였다. C. indica 균사체의 효율적 배양을 위한 조건은 최적 배양 온도 30-32℃, pH 6.0, 탄소원 maltose 2%, 유기질소원 yeast extract 1%, 무기질소원 NaNO3 0.1%, 아미노산 asparagine 0.7%, 유기산 acetic acid 0.1%, 무기염류 MnSO4 0.7 mM가 최적 배양조건 이었다. 새로이 MYNA로 명명한 C. indica의 최적 배지는 기존의 PDA 배지보다 균사의 생장이 균일하고, PDA 배지의 배양기간과 비교하였을 때 10.5% 단축하는 효과가 있었다. MYNA 배지는 추후 C. indica의 재배에 널리 활용될 수 있을 것으로 판단된다.

췌장베타세포에서 스트렙토초토신으로 유도한 당뇨병 실험 모델에 대한 팔미원의 영향 (Effect of Palmiwon on the Streptozotocin induced Prediabetic Model in Panceratic Bita Cells)

  • 이인순;이인자
    • Biomolecules & Therapeutics
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    • 제6권4호
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    • pp.371-377
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    • 1998
  • The aim of the present study was to investigate the effect of Palmiwon on the type 1-prediabetic models induced by streptozotocin (STZ) in RINm5F cells and HIT-T15 cells. Palmiwon increased the cell proliferation and insulin release when pre- and post-treated for the STZ-exposed pancreatic beta cells. The cell proliferation and insulin release of these beta cells were measured by $^3$H-thymidine uptake and RIA, respectively. We also analyzed nutrients such as sugars, fatty acid and amino acids and minerals containing in Palmiwon using by gas chromatography, amino acid analyzer and AA spectrometer, respectively. Palmiwon seems to have protective and recovery properties on the prediabetic model in cellular level, which were ascribe to various nutrients and minerals containing in Palmiwon. From these results, it could be suggested that Palmiwon may be available as preventive and therapeutic prescription of type 1 diabetes mellitus.

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Effects of Replacing Spray Dried Plasma Protein With Spray Dried Porcine Intestine Hydrolysate on Ileal Digestibility of Amino Acids and Growth Performance in Early-Weaned Pigs

  • Kim, J.H.;Chae, B.J.;Kim, Y.G.
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권12호
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    • pp.1738-1742
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    • 2000
  • A study was conducted to determine the ileal digestibility (ID) of amino acids and feeding values of spray dried plasma protein (SDPP) and spray dried porcine intestine hydrolysate (SDPI) in early-weaned pigs. Twelve pigs aged 18 days old (Landrace${\times}$Yorkshire${\times}$Duroc; $5.83{\pm}0.51kg$ BW) were cannulated in the terminal ileum for determination of ID of amino acids. Ninety pigs ($6.28{\pm}0.1kg$, 18 days old) were also employed for a feeding trial during phase I period. Treatments were: 1) 6% SDPP, 2) 6% SDPI, and 3) 3% SDPP+3% SDPI. The apparent and true ID values of the essential amino acids except leucine, methionine and valine were lower (p<0.01) in SDPI than in SDPP. The average apparent ID of essential amino acids in SDPP and SDPI were 75.63 and 71.30%, and the average true ID of essential amino acids 84.83 and 80.51%, respectively. The ADG and feed conversion ratio in piglets fed the 6% SDPP diet were better (p<0.01) than in those fed the 6% SDPI diet. When 3% of SDPP was replaced by SDPI, however, the growth rate and efficiency of pigs were comparable to those in pigs fed 6% SDPP. In conclusion, SDPP can be partially replaced by SDPI without any detrimental effect on growth performance in early-weaned pigs.

Zn2+ PVC-based Membrane Sensor Based on 3-[(2-Furylmethylene)amino]-2-thioxo-1,3-thiazolidin-4-one

  • Ganjali, Mohammad Reza;Zamani, Hassan Ali;Norouzi, Parviz;Adib, Mehdi;Rezapour, Morteza;Aceedy, Mohammad
    • Bulletin of the Korean Chemical Society
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    • 제26권4호
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    • pp.579-584
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    • 2005
  • The 3-[(2-furylmethylene)amino]-2-thioxo-1,3-thiazolidin-4-one (FTT) was used as an excellent ionophore in construction of a $Zn^{2+}$ PVC-based membrane sensor. The best performance was obtained with a membrane composition of 30% poly(vinyl chloride), 62% nitrobenzen (NB), 3% FTT and 5% sodium tetraphenyl borate (TBP). This membrane sensor shows very good selectivity and sensitivity towards $Zn^{2+}$ over a wide variety of cations, including alkali, alkaline earth, transition and heavy metal ions. The membrane sensor revealed a great enhancement in selectivity coefficients for $Zn^{2+}$ ions, in comparison to the previously reported $Zn^{2+}$ membrane sensors. Theoretical studies also showed the selective interaction of TFF and $Zn^{2+}$ ions. The proposed membrane sensor exhibits a Nernstian behavior (with slope of 29.3 ${\pm}$ 0.3 mV per decade) over a wide concentration range (1.0 ${\times}$ $10^{-6}$-1.0 ${\times}$ $10^{-2}$) with a detection limit of 8.5 ${\times}$ $10^{-7}$ M (52 ng mL$^{-1}$). It shows relatively fast response time, in the whole concentration range ($\lt$ 20 s), and can be used for at least 10 weeks in a pH range of 3.0-7.0. The proposed membrane sensor was successfully used in direct determination of $Zn^{2+}$ ions in wastewater of industrial zinc electroplating companies, and also as an indicator electrode in titration with EDTA.

세자리 Schiff Base의 Ni(II) 착물의 분석 (Analysis of Tridentate Schiff Base Ni(II) Complex)

  • 채희남;최용국
    • 분석과학
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    • 제11권5호
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    • pp.332-340
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    • 1998
  • Salicylaldehyde와 2-hydroxy-1-naphthaldehyde를 2-aminophenol과 2-amino-p-cresol에 반응시켜 세자리 Schiff base 리간드($SIPH_2$, $SIPCH_2$, $HNIPH_2$, $HNIPCH_2$)들을 합성하였다. 이들 리간드를 Ni(II) 이온과 반응시켜 세자리 Schiff base Ni(II) 착물들을 합성 하였다. 이들 리간드와 그 착물들의 가상적인 구조와 특성을 원소분석, $^1H$-NMR, IR, UV-vis 분광법과 열 무게 분석법으로 알아보았다. Schiff base 리간드와 Ni(II) 착물의 몰비는 1:1로 결합하며, Ni(II)착물들은 3분자의 수화물이 배위된 6배위의 8면체 구조임을 알았다. 지지 전해질로서 0.1 M TBAP를 포함한 DMSO 용액에서 순환 전압전류법과 미분 펄스 전압전류법으로 세자리 Schiff base 리간드와 이들의 Ni(II) 착물들의 전기 화학적인 산화 환원 과정을 알아보았다. 세자리 Schiff base 리간드들의 전기 화학적 환원은 확산 지배적이고 비가역적으로 진행되었다. Ni(II) 착물의 전기화학적 환원과정은 1단계 1전자 반응으로 확산 지배적이고 준가역적으로 진행되었다. Ni(II) 착물들의 환원전위는 [$Ni(II)(HNIP)(H_2O)_3$]>[$Ni(II)(SIP)(H_2O)_3$]>[$Ni(II)(SIPC)(H_2O)_3$]>[$Ni(II)(HNIPC)(H_2O)_3$]순으로 양전위 방향으로 이동하였으며, 리간드의 영향은 크게 받지 않았다. 이들 결과로 부터 본 연구에서 합성한 Ni(II)착물은 [$Ni(II)(HNIPC)(H_2O)_3$] 착물이 DMSO용매에서 가장 안정함을 알 수 있다.

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순무의 이화학적 및 기능적 특성 (Physicochemical and Functional Properties of Turnip)

  • 박용곤;김흥만;박미원;김성란;최인욱
    • 한국식품영양과학회지
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    • 제28권2호
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    • pp.333-341
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    • 1999
  • This study was conducted to investigate physicochemical and functional properties of turnip. The concentrations of antocyanin in the rind of root was 3.24mg%, which was about 3.5 times higher than those in the flesh. Among free amino acids in each part of turnip, glutamic acid showed the highest concentrations(132.5mg%) in the root followed by alanine(25.0mg%), valine (23.3mg%), and serine (20.7mg%). Contents of minerals in the root of turnip were 395.7mg% for potassium, 187mg% for calcium, 53.6mg% for phosphate, and 40.7mg% for magnesium. According to dynamic heasdspace analysis, dimethyl disulfide was the most abundant flavor components from the juice of fresh turnip and other volatile components such as dimethyl trisulfide, 1 hexanal, 2 methyl 1 butamine, and 1 penten 3 ol were also identified. From the blanched turnip, however, 3 isothiocyanato 1 propene and 4 isothiocyanato 1 butene were mainly detected. Fresh and blanched turnips were extracted with water or 70% acetone to investigate percent yield. The concentrations of total polyphenols in the extracts of fresh turnip were higher than those of blanched turnip. The nitrite scavenging effects of water and 70% acetone extracts from fresh or blanched turnip were decreased as pH was increased. Between two water extracts, the 30oC water extracts showed the higher values of electron donating ability than 95$^{\circ}C$ water extracts.

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