• Title/Summary/Keyword: L-Histidine

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Purification and Characterization of a Thermostable ${\beta}-Glycosidase$ from Thermus caldophilus GK24

  • Yoo, Jin-Sang;Han, Ki-Woong;Kim, Hyun-Kyu;Kim, Min-Hong;Kwon, Suk-Tae
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.638-642
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    • 2000
  • A ${\beta}-glycosidase$ enzyme with $\beta$-D-fucosidase, ${\beta}-D-galactosidase$, and $\beta$-D-glucosidase activities has been purified from Thermus caldophilus GK24. The enzyme was monomeric with a molecular mass of 49 kDa, as evidenced by SDS-PAGE. The $K_m$ values for p-nitrophenyl ${\beta}-D-fucopyranoside$ (p-NPFuc), p-nitrophenyl ${\beta}-D-galactopyranoside$ (p-NPGal), and p-nitrophenyl ${\beta}-D-glucopyranoside$ (p-NPGlu) were 0.23 mM, 6.25 mM, and 0.28 mM, respectively. The enzyme showed optimal pH ranging between 5.5-6.5 and maximum temperature in the range of $85-90^{\circ}C$ for all the above mentioned activities. The half-life of the enzyme in sodium phosphate buffer (pH 6.0) at $80^{\circ}C$ was approximately 7 h. The p-NPGal hydrolyzing activity of Tca ${\beta}-glycosidase$ was strongly activated by L-histidine, while the p-NPFuc and p-NPGlu hydrolyzing activities of Tca ${\beta}-glycosidase$ were not affected at all by the amino acid. These results suggest differences in the conformation or in the reactive residues at the active site of Tca ${\beta}-glycosidase$.

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Studies on the Charge-transfer Complex including Aflatoxin $B_1$ -Part I. Charge-transfer Complex with Benzene- (Aflatoxin $B_1$ Charge-transfer Complex에 관(關)한 연구(硏究) -제1보(第一報) Benzene과의 Charge-transfer Complex-)

  • Noh, Ick-Sam;Lee, Kang-Heup
    • Applied Biological Chemistry
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    • v.17 no.2
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    • pp.143-148
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    • 1974
  • The interaction of the carcinogenic mycotoxin, Aflatoxin $B_1$, with the electron-donating molecule, benzene, was studied spectrophotometrically. The formation of charge-transfer complex between Aflatoxin $B_1$ and benzene in the presence of zinc chloride was observed and the apparent equilibrium constant of this charge-transfer complex was found to be 0.198 (liter $mole^{-1}$). It is assumed that, as the result of this study, some charge-transfer complexes could be formed between the weak electron-accepting Aflatoxin $B_1$ and strong electron-donating molecules, and the spectral changes occurred in the binding of Aflatoxin $B_1$ with proteins or DNA is attributed to the existence of charge-transfer type interaction.

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Antioxidant and ACE Inhibiting Activities of the Rockfish Sebastes hubbsi Skin Gelatin Hydrolysates Produced by Sequential Two-step Enzymatic Hydrolysis

  • Kim, Hyung-Jun;Park, Kwon-Hyun;Shin, Jun-Ho;Lee, Ji-Sun;Heu, Min-Soo;Lee, Dong-Ho;Kim, Jin-Soo
    • Fisheries and Aquatic Sciences
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    • v.14 no.1
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    • pp.1-10
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    • 2011
  • This study was conducted to obtain hydrolysates with potent antioxidative activity from rockfish skin gelatin. Gelatin was extracted under high temperature/high pressure using a two-step enzymatic hydrolysis with commercial enzymes such as Alcalase, Flavourzyme, Neutrase, and Protamex. The second rockfish-skin gelatin hydrolysate (SRSGH) was prepared by further incubating the first gelatin hydrolysate (FRSGH), which had been hydrolyzed with Alcalase for 1-h (FRSGH-A1), with Flavourzyme for 2-h (SRSGH-F2). The second gelatin hydrolysate showed higher antioxidative activity of 3.72 as measured by a Metrohm Rancimat and superior angiotensin I-converting enzyme (ACE) inhibiting activity of 0.82 mg/mL. Compared with the gelatin, the relative proportion in SRSGH-F2 was markedly decreased in the 100-kDa peak, whereas it was increased in that less than 100-kDa. The amino acid composition of SRSGH-F2 was rich in glycine (25.9%), proline (10.8%), alanine (9.1%), and glutamic acid (9.1%). In contrast, it was poor in cystine (not detected), methionine (1.6%), tyrosine (0.4%), hydroxylysine (0.9%), and histidine (0.9%). In recent years, demand for natural functional foods has been increasing, and SRSGH-F2 can be used as a functional food ingredient in the food industries. However, further detailed studies on SRSGH-F2 with regard to its antioxidant activity in vivo and the various antioxidant mechanisms are needed.

Effect of Dietary Supplementation of Blood Meal and Additional Magnesium on Carnosine and Anserine Concentrations of Pig Muscles

  • Park, Se Won;Kim, Chan Ho;Kim, Jong Woong;Shin, Hye Seong;Paik, In Kee;Kil, Dong Yong
    • Food Science of Animal Resources
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    • v.34 no.2
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    • pp.252-256
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    • 2014
  • The objective of this study was to investigate the effect of dietary supplementation of blood meal as a source of L-histidine, and the addition of magnesium (Mg) as a catalyst of carnosine synthetase for the carnosine and anserine concentrations of pig muscles (longissimus dorsi, LD and vastus intermedius, VI). A total of twenty-four pigs with an average body weight of $60.2{\pm}4.2$ kg were randomly allotted to one of three dietary treatments (eight replicates), during 56 d of the feeding trial. Dietary treatments included: (1) Basal: basal diet; (2) BM: 95% basal diet + 5% blood meal; and (3) BM+Mg: 94.8% basal diet + 5% blood meal + 0.2% MgO (60% Mg). Results indicated that drip loss in the LD was less (p<0.05) for meat with BM+Mg treatment than that with Basal treatment, but the values for BM treatment did not differ from those of the other two treatment groups. The concentrations of carnosine in the LD were increased by 10.0% in both BM and BM+Mg treatment groups over the Basal treatment group (significance not verified). The concentrations of carnosine and anserine in the VI were not affected by the dietary treatments. Inclusion of additional Mg in diets had no effect on carnosine and anserine concentrations in the LD and VI. In conclusion, dietary supplementation of blood meal could be a potential method of fortifying the pork with carnosine. Inclusion of additional Mg in the diets containing blood meal had no benefit on carnosine and anserine depositions in pig muscles.

Composition of Free Amino Acids and Essential Oils in Root of Anthriscus sylvestylis (전조 뿌리의 유리 아미노산과 정유 성분 조성)

  • 김상국;권태용;민기군;이승필;최부술;이상철
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.5
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    • pp.521-525
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    • 1996
  • The study was carried out to find compositions of proximate components, free amino acid, and essential oils from root of Anthriscus sylvestylis. Proximate component contents were 7.69% for protein, 1.74% for fat, 2.44% for fiber, and 3.76% for ash. Extract content was 27.68% in fresh root. The compositions of free amino acids consisted 16 kinds. Phenylalanine content was the highest in composition of free amino acids. The essential oils of the root of Anthriscus sylvestylis was examined. $\alpha$-pinene, campreol, ,$\beta$-pinene, sabinene, myrcene, phellandrene, $\alpha$-terpinolene, d-limone, ${\gamma}$-terpinene, p-cymene, $\alpha$-terpinolene, carboxaldehyde, 3-cyc1ohexen-l-carboxaldehyde, 2-nonenal, isobornyl acetate, 4-terpineol, $\beta$-bisabolene, cis-piperitol, p-cymen-8-ol, BHT, methyl eugenol and 2-methoxy-4-vinyl-phenol were identified from the diethylether layers. Recovery yield of essential oils of Anthriscus sylvestylis of root was 0.58%. As a result, it was considered that the plant is worthy of cultivating as spice and medicinal crops.

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Biological Active Substance Produced by a Strain of Streptomyces sp. (Part.III) Purification and Nutritional Requirement. (Streptomyces 속 균주가 생성한 물질의 생물활성 (제삼보) 정제 및 영양요구성)

  • 송방호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.5 no.1
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    • pp.36-45
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    • 1977
  • A piscicidal substance was isolated from the culture medium of Streptomyces umbrosus by avicel column chromatography and avicel thin layer chromatography after extration with chroloform. Bluegreen fluorescence was emitted under UV irradiation. Factors which govern toxin production and nutrition requirement for high toxin titres were observed. Nutritional uptake for toxin production was not curresponded with that for cell growth. Alanine, valine, serine asparagine, arginine, histidine, urea and sodium nitrate as a carbon source and glucose, mannose, rhamnose, xylose, arabitol and starch as a carbon source were recognized as a favorable nutrient for high toxin production. Magnesium was essential factor whereas vitamins were not of effective. Most of toxin was formed simultaneously with cell growth in esponential phase. Maximal production was observed for six day culture at 3$0^{\circ}C$. Tissues of gill, kidney and pnacreas in Cyprinus carpio were denatured extreamly after treating with the substance. Atrophied nucleous, indented membrane and degradated cytoplasm with necrotic affectness were noted on each tissue. The chemical formula of the substance was designated as $C_{38}$ $H_{66}$ $NO_4$.

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Cloning and Expression of a Thermostable ${\alpha}$-Galactosidase from the Thermophilic Fungus Talaromyces emersonii in the Methylotrophic Yeast Pichia pastoris

  • Simila, Janika;Gernig, Anita;Murray, Patrick;Fernandes, Sara;Tuohy, Maria G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1653-1663
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    • 2010
  • The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

Effects of a pineapple (Ananas comosus L.) cannery by-product on growth performance and carcass characteristics in finishing Hanwoo steers

  • Choi, Yongjun;Lee, Sangrak;Na, Youngjun
    • Animal Bioscience
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    • v.34 no.2
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    • pp.233-242
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    • 2021
  • Objective: The aim of this study was to determine the effect of pineapple cannery by-product (PCB) level on the growth performance and carcass characteristics of finishing Hanwoo steers. Methods: The feeding stage was divided into early and late finishing stages. A total of 60 castrated Hanwoo steers (13.9±0.8 months old, 418.8±36.5 kg initial body weight [BW]) were blocked by initial BW and then randomly allotted into 12 pens (five head/pen). The pens were randomly assigned to control (CONT), low PCB (LPCB), or high PCB (HPCB) treatments. These diets contained 0%, 1.5%, or 3.0% of PCB (on a dry matter [DM] basis; as-fed basis was 0%, 10.6%, or 21.2%), respectively. Results: For the early finishing stage, body weight gain (BWG) and average daily gain (ADG) of the CONT and LPCB feeding groups were greater (p<0.05) than those of the HPCB feeding group. In addition, there were linear and quadratic effects on BWG and ADG with increasing dietary PCB level (p<0.05). The gain to feed (G:F) ratio tends to quadratically decrease with an increasing PCB level in the early finishing stage (p = 0.076). Growth performances of late finishing stage were not affected by PCB level. The marbling score of the LPCB feeding group was similar to that of the CONT feeding group. However, there was a linear decrease (p< 0.05) in marbling score and quality grade among treatments as PCB was increased in the diet. In the longissimus muscle free amino acid profile, histidine composition increased linearly (p<0.05) with an increasing level of PCB. Conclusion: The level of PCB 1.5% DM in diet can be used for finishing steers without any adverse effects on growth and carcass performances. However, there were some negative effects on growth and carcass performance in the HPCB feeding group.

T-Cell Death-Associated Gene 51 Is a Novel Negative Regulator of PPARγ That Inhibits PPARγ-RXRα Heterodimer Formation in Adipogenesis

  • Kim, Sumi;Lee, Nari;Park, Eui-Soon;Yun, Hyeongseok;Ha, Tae-Uk;Jeon, Hyoeun;Yu, Jiyeon;Choi, Seunga;Shin, Bongjin;Yu, Jungeun;Rhee, Sang Dal;Choi, Yongwon;Rho, Jaerang
    • Molecules and Cells
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    • v.44 no.1
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    • pp.1-12
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    • 2021
  • The nuclear receptor peroxisome proliferator-activated receptor γ (PPARγ) is the master transcriptional regulator in adipogenesis. PPARγ forms a heterodimer with another nuclear receptor, retinoid X receptor (RXR), to form an active transcriptional complex, and their transcriptional activity is tightly regulated by the association with either coactivators or corepressors. In this study, we identified T-cell death-associated gene 51 (TDAG51) as a novel corepressor of PPARγ-mediated transcriptional regulation. We showed that TDAG51 expression is abundantly maintained in the early stage of adipogenic differentiation. Forced expression of TDAG51 inhibited adipocyte differentiation in 3T3-L1 cells. We found that TDAG51 physically interacts with PPARγ in a ligand-independent manner. In deletion mutant analyses, large portions of the TDAG51 domains, including the pleckstrin homology-like, glutamine repeat and proline-glutamine repeat domains but not the proline-histidine repeat domain, are involved in the interaction with the region between residues 140 and 506, including the DNA binding domain, hinge, ligand binding domain and activation function-2 domain, in PPARγ. The heterodimer formation of PPARγ-RXRα was competitively inhibited in a ligand-independent manner by TDAG51 binding to PPARγ. Thus, our data suggest that TDAG51, which could determine adipogenic cell fate, acts as a novel negative regulator of PPARγ by blocking RXRα recruitment to the PPARγ-RXRα heterodimer complex in adipogenesis.

Effects of rumen-protected amino acid prototypes on rumen fermentation characteristics in vitro

  • Gyeongjin, Kim;Tabita Dameria, Marbun;Jinhyun, Park;Sang Moo, Lee;Hong Gu, Lee;Jun Ok, Moon;Jin Seung, Park;Eun Joong, Kim
    • Korean Journal of Agricultural Science
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    • v.48 no.4
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    • pp.669-679
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    • 2021
  • This study was conducted to evaluate the effects of rumen-protected amino acid (RPAA) prototypes, which were chemically synthesized, on in vitro rumen fermentation and protection rate outcomes. Several RPAA prototypes were incubated with timothy hay and concentrate. Treatments consisted of 1) control (CON; no RPAA prototype supplement), and prototypes of 2) 0.5% RP-methionine (RPMet), 3) 0.5% RP-tryptophan (RPTrp), 4) 0.5% RP-valine (RPVal), 5) 0.5% RP-phenylalanine (RPPhe), 6) 0.5% RP-leucine (RPLeu), 7) 0.5% RP-histidine (RPHis), 8) 20% RPMet, and 9) 20% RPTrp (w·w-1 feed). The inoculum (50 mL) prepared with rumen fluid and McDougall's buffer (1 : 4) was dispensed in individual serum bottles and was anaerobically incubated for 0, 6, and 24 h at 39℃ in triplicate. The dry matter degradability did not differ among the groups, except for the 20% RPMet and the 20% RPTrp treatments at 6 and 24 h. The total volatile fatty acid concentration in the 20% RPMet was higher (p < 0.05) than the rest of the groups at 6 h, and 20% RPMet showed the highest molar proportion of acetate, whereas the lowest proportion of propionate was found at 6 h (p < 0.05). The protection rate of the RPAA prototypes ranged from 29.85 to 109.21%. at 24 h. In conclusion, the chemically synthesized RPAA prototypes studied here had no detrimental effects on rumen fermentation parameters. Further studies using animal models are needed for more accurate evaluations of the effectiveness of RPAA.