• 제목/요약/키워드: GH9

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Molecular Characterization of the α-Galactosidase SCO0284 from Streptomyces coelicolor A3(2), a Family 27 Glycosyl Hydrolase

  • Temuujin, Uyangaa;Park, Jae Seon;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제26권9호
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    • pp.1650-1656
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    • 2016
  • The SCO0284 gene of Streptomyces coelicolor A3(2) is predicted to encode an α-galactosidase (680 amino acids) belonging to glycoside hydrolase family 27. In this study, the SCO0284 coding region was cloned and overexpressed in Streptomyces lividans TK24. The mature form of SCO0284 (641 amino acids, 68 kDa) was purified from culture broth by gel filtration chromatography, with 83.3-fold purification and a yield of 11.2%. Purified SCO0284 showed strong activity against p-nitrophenyl-α-D-galactopyranoside, melibiose, raffinose, and stachyose, and no activity toward lactose, agar (galactan), and neoagarooligosaccharides, indicating that it is an α-galactosidase. Optimal enzyme activity was observed at 40℃ and pH 7.0. The addition of metal ions or EDTA did not affect the enzyme activity, indicating that no metal cofactor is required. The kinetic parameters Vmax and Km for p-nitrophenyl-α-D-galactopyranoside were 1.6 mg/ml (0.0053 M) and 71.4 U/mg, respectively. Thin-layer chromatography and mass spectrometry analysis of the hydrolyzed products of melibiose, raffinose, and stachyose showed perfect matches with the masses of the sodium adducts of the hydrolyzed products, galactose (M+Na, 203), melibiose (M+Na, 365), and raffinose (M+Na, 527), respectively, indicating that it specifically cleaves the α-1,6-glycosidic bond of the substrate, releasing the terminal D-galactose.

Cloning of Agarase Gene from Non-Marine Agarolytic Bacterium Cellvibrio sp.

  • Ariga, Osamu;Inoue, Takayoshi;Kubo, Hajime;Minami, Kimi;Nakamura, Mitsuteru;Iwai, Michi;Moriyama, Hironori;Yanagisawa, Mitsunori;Nakasaki, Kiyohiko
    • Journal of Microbiology and Biotechnology
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    • 제22권9호
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    • pp.1237-1244
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    • 2012
  • Agarase genes of non-marine agarolytic bacterium Cellvibrio sp. were cloned into Escherichia coli and one of the genes obtained using HindIII was sequenced. From nucleotide and putative amino acid sequences (713 aa, molecular mass; 78,771 Da) of the gene, designated as agarase AgaA, the gene was found to have closest homology to the Saccharophagus degradans (formerly, Microbulbifer degradans) 2-40 aga86 gene, belonging to glycoside hydrolase family 86 (GH86). The putative protein appears to be a non-secreted protein because of the absence of a signal sequence. The recombinant protein was purified with anion exchange and gel filtration columns after ammonium sulfate precipitation and the molecular mass (79 kDa) determined by SDS-PAGE and subsequent enzymography agreed with the estimated value, suggesting that the enzyme is monomeric. The optimal pH and temperature for enzymatic hydrolysis of agarose were 6.5 and $42.5^{\circ}C$, and the enzyme was stable under $40^{\circ}C$. LC-MS and NMR analyses revealed production of a neoagarobiose and a neoagarotetraose with a small amount of a neoagarohexaose during hydrolysis of agarose, indicating that the enzyme is a ${\beta}$-agarase.

성장장애 흰쥐에서 경옥고(瓊玉膏)와 경옥고가록용(瓊玉膏加鹿茸)의 성장효과 비교 연구 (A Comparative Study on Effects of Kyungohkgo and Kyungohkgo Ga Nokyong on Growth in Growth Deficiency Rat with Insufficient Nutrition Diet)

  • 차윤엽
    • 한방비만학회지
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    • 제9권1호
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    • pp.59-69
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    • 2009
  • Objectives This study was designed to evaluate the difference of between Kyungohkgo and Kyungohkgo Ga Nokyong effects of growth Methods We divided male Spraque-Dawley rats into 6 groups. They were Normal group, Growth deficiency rat with insufficient nutrition diet group, Growth deficiency rat with 0.1% Kyungohkgo group, Growth deficiency rat with 0.2% Kyungohkgo group, Growth deficiency rat with 0.1% Kyungohkgo Ga Nokyong group and Growth deficiency rat with 0.2% Kyungohkgo Ga Nokyong group. They were administered for 5 weeks. We measured body weight, serum growth hormone, insulin-like growth factor and thyroid stimulating hormone, RBC, concentration of Hb and PCV ratio, total WBC and its composition, the values of plasma glutamic oxaloacetic transaminase(GOT) and glutamic pyruvic transaminase(GPT) activities. Results 1. In body weight, Kyungohkgo 0.1%, Kyungohkgo Ga Nokyong 0.1%, 0.2% groups were showed sighnificantly different than control group. But That groups were not showed significantly different than each others. 2. In serum growth hormone, insulin-like growth factor and thyroid stimulating hormone, we obtained the results of tendency to increase in Kyungohkgo Ga Nokyong group, however these values showed no significantly different. 3. In the counts of RBC, Kyungohkgo group and Kyungohkgo Ga Nokyong group were showed significantly different than control group. But these values showed no significantly different. In the concentration of Hb, Kyungohkgo Ga Nokyong 0.2%group was showed significantly different than control group. Kyungohkgo Ga Nokyong 0.2% group and Kyungohkgo 0.2% group were showed significantly different than control group. 4. The counts of total WBC and its composition showed no significantly different in all treatment groups. 5. The values of plasma glutamic oxaloacetic transaminase(GOT) and glutamic pyruvic transaminase(GPT) activities showed no significantly different in all treatment groups. Conclusions So Kyungohkgo Ga Nokyong and Kyungohkgo have an effect of promoting growth of rats. And We know that the effect of Kyungohkgo Ga Nokyong is better than Kyungohkgo.

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Effects of Active Immunization against Somatostatin or its Analogues on Milk Protein Synthesis of Rat Mammary Gland Cells

  • Kim, J.Y.;Cho, K.K.;Chung, M.I.;Kim, J.D.;Woo, J.H.;Yun, C.H.;Choi, Y.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권4호
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    • pp.570-575
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    • 2002
  • Effects of active immunization against native 14-mer somatostatin (SRIF, somatotropin releasing inhibiting factor) and its two 14-mer-somatostatin analogues on the milk production in rat mammary cells were studied. Native SRIF, Tyr11-somatostatin (Tyr11-SRIF), and D-Trp8, D-Cys14-somatostatin (Trp8Cys14-SRIF) were conjugated to bovine serum albumin (BSA) for immunogen preparation. Twenty-four female Sprague-Dawley rats were divided into four groups and immunized against saline (Control), SRIF, Tyr11-SRIF, and Trp8Cys14-SRIF at five weeks of age. Booster immunizations were performed at 7, 9, and 11 weeks of age. SRIFimmunized rats were mated at 10 weeks of age. The blood and mammary glands were collected at day 15 post-pregnancy and -lactation. To measure the amount of milk protein synthesis in the mammary gland, mammary cells isolated from the pregnant and the lactating rats, were cultured in the presence of $^3H$-lysine. No significant differences in growth performance, concentration of growth hormone in the circulation, and the amount of milk protein synthesis were observed among the groups. Inductive levels of serum anti-SRIF antibody in the SRIF and Tyr11-SRIF groups but not in the Trp8Cys14-SRIF group, were significantly higher than that of the control group during the pregnancy and lactation periods. The result suggests that active immunization against native 14-mer SRIF and Tyr11-SRIF was able to induce anti-SRIF antibodies, but did not affect the milk protein synthesis.

DdeI Polymorphism in Coding Region of Goat POU1F1 Gene and Its Association with Production Traits

  • Lan, X.Y.;Pan, C.Y.;Chen, H.;Lei, C.Z.;Hua, L.S.;Yang, X.B.;Qiu, G.Y.;Zhang, R.F.;Lun, Y.Z.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권9호
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    • pp.1342-1348
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    • 2007
  • POU1F1 is a positive regulator for GH, PRL and TSH${\beta}$ and its mutations associate with production traits in ruminant animals. We described a DdeI PCR-RFLP method for detecting a silent allele in the goat POU1F1 gene: TCT (241Ser)>TCG (241Ser). Frequencies of $D_1$ allele varied from 0.600 to 1.000 in Chinese 801 goats. Significant associations of DdeI polymorphism with production traits were found in milk yield (*p<0.05), litter size (*p<0.05) and one-year-old weight (*p<0.05) between different genotypes. Individuals with genotype $D_1D_1$ had a superior performances when compared to those with genotype $D_1D_2$ (*p<0.05). Hence, the POU1F1 gene was suggested to the potential candidate gene for superior milk performance, reproduction trait and weight trait. Genotype $D_1D_1$, characterized by a DdeI PCR-RFLP detection, was recommended to geneticists and breeders as a molecular marker for better performance in the goat industry.

The Membrane-Bound NADH:Ubiquinone Oxidoreductase in the Aerobic Respiratory Chain of Marine Bacterium Pseudomonas nautica

  • Lee, Young-Jae;Cho, Kyeung-Hee;Kim, Young-Jae
    • Journal of Microbiology and Biotechnology
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    • 제13권2호
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    • pp.225-229
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    • 2003
  • Each oxidoreductase activity of the aerobic respiratory chain-linked NADH oxidase system in the marine bacterium Pseudomonas nautica was stimulated by monovalent cations including $Na^+,\;Li^+,\;and\;K^+$. In the presence of NADH or deamino-NADH as electron donors, $GH_2$ formation was approximately 1.3-fold higher in the presense of 0.08 M of $Na^+\;than\;K^+$, Whereas the other reductase activities were not significantly higher in $Na^+\;than\;K^+$. The optimal pH of NADH (or deamino-NADH):ubiquinone-1 oxidoreductase was 9.0 in the presence of 0.08 M NaCl. The activity of NADH (or deamino-NADH):ubiquinone-1 oxidoreductase was inhibited by about 33% with $60{\mu}M$ 2-heptyl-4-hydroxyquinoline-N-oxide (HQNO). The activity of NADH (deamino-NADH): ubiquinone-1 oxidoreductase was inhibited by about 32 to 38% with $80{\mu}M$ rotenone, whereas the activity was highly resistant to capsaicin. On the other hand, electron transfer from NADH or deamino-NADH to ubiquinone-1 generated a membrane potential (${\Delta}{\psi}$) which was larger in the presence of $Na^+$ than that observed in the absence of $Na^+$. The ${\Delta}{\psi}$ was almost completely collapsed by $5{\mu}M$ carbonylcyanide m-chlorophenylhydrazone(CCCP), and approximately 50% inhibited by $100{\mu}M$ rotenone, or $60{\mu}M$ 2-heptyl-4-hydroxyquinoline (HQNO). Also, HQNO made the ${\Delta}{\psi}$ very unstable. The results suggest that the enzymatic and energetic properties of the NADH:ubiquinone oxidoreductase of P. nautica are quite different, compared with those of other marine halophilic bacteria.

Evaluation of Ethanol Production Activity by Engineered Saccharomyces cerevisiae Fermenting Cellobiose through the Phosphorolytic Pathway in Simultaneous Saccharification and Fermentation of Cellulose

  • Lee, Won-Heong;Jin, Yong-Su
    • Journal of Microbiology and Biotechnology
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    • 제27권9호
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    • pp.1649-1656
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    • 2017
  • In simultaneous saccharification and fermentation (SSF) for production of cellulosic biofuels, engineered Saccharomyces cerevisiae capable of fermenting cellobiose has provided several benefits, such as lower enzyme costs and faster fermentation rate compared with wild-type S. cerevisiae fermenting glucose. In this study, the effects of an alternative intracellular cellobiose utilization pathway-a phosphorolytic pathway based on a mutant cellodextrin transporter (CDT-1 (F213L)) and cellobiose phosphorylase (SdCBP)-was investigated by comparing with a hydrolytic pathway based on the same transporter and an intracellular ${\beta}$-glucosidase (GH1-1) for their SSF performances under various conditions. Whereas the phosphorolytic and hydrolytic cellobiose-fermenting S. cerevisiae strains performed similarly under the anoxic SSF conditions, the hydrolytic S. cerevisiae performed slightly better than the phosphorolytic S. cerevisiae under the microaerobic SSF conditions. Nonetheless, the phosphorolytic S. cerevisiae expressing the mutant CDT-1 showed better ethanol production than the glucose-fermenting S. cerevisiae with an extracellular ${\beta}$-glucosidase, regardless of SSF conditions. These results clearly prove that introduction of the intracellular cellobiose metabolic pathway into yeast can be effective on cellulosic ethanol production in SSF. They also demonstrate that enhancement of cellobiose transport activity in engineered yeast is the most important factor affecting the efficiency of SSF of cellulose.

Fabrication, characterization, simulation and experimental studies of the ordinary concrete reinforced with micro and nano lead oxide particles against gamma radiation

  • Mokhtari, K.;Kheradmand Saadi, M.;Ahmadpanahi, H.;Jahanfarnia, Gh.
    • Nuclear Engineering and Technology
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    • 제53권9호
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    • pp.3051-3057
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    • 2021
  • The concrete is considered as an important radiation shielding material employed widely in nuclear reactors, particle accelerators, laboratory hot cells and other different radiation sources. The present research is dedicated to the shielding properties study of the ordinary concrete reinforced with different weight fractions of lead oxide micro/nano particles. Lead oxide particles were fabricated by chemical synthesis method and their properties including the average size, morphological structure, functional groups and thermal properties were characterized by XRD, FESEM-EDS, FTIR and TGA analysis. The gamma ray mass attenuation coefficient of concrete composites has been calculated and measured by means of the Monte Carlo simulation and experimental methods. The simulation process was based on the use of MCNP Monte Carlo code where the mass attenuation coefficient (μ/ρ) has been calculated as a function of different particle sizes and filler weight fractions. The simulation results showed that the employment of the lead oxide filler particles enhances the mass attenuation coefficient of the ordinary concrete, drastically. On the other hand, there are approximately no differences between micro and nano sized particles. The mass attenuation coefficient was increased by increasing the weight fraction of nanoparticles. However, a semi-saturation effect was observed at concentrations more than 10 wt%. The experimental process was based on the fabrication of concrete slabs filled by different weight fractions of nano lead oxide particles. The mass attenuation coefficients of these slabs were determined at different gamma ray energies using 22Na, 137Cs and 60Co sources and NaI (Tl) scintillation detector. The experimental results showed that the HVL parameter of the ordinary concrete reinforced with 5 wt% of nano PbO particles was reduced by 64% at 511 keV and 48% at 1332 keV. Reasonable agreement was obtained between simulation and experimental results and showed that the employment of nano PbO particles is more efficient at low gamma energies up to 1Mev. The proposed concrete is less toxic and could be prepared in block form instead of toxic lead blocks.

농약 품목에 대한 GHS 기준과 농진청 기준에 따른 자극성 산출 후 자극성 분류 차이 비교 (Comparison of Differences between the Results of Irritation Classification after Irritation Calculation with GHS Criteria and RDA Directives in Plant Protection Products)

  • 유아선;오진아;박수진;조유미;이제봉;이난희;이주연;임양빈
    • 농약과학회지
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    • 제20권4호
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    • pp.326-340
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    • 2016
  • 농약의 원제에 대한 GHS 체계 도입에 따라 농약 품목에 대한 GHS 도입 필요성이 대두되고 있어 GHS 도입 시 현행과의 평가 기준을 비교하였다. 농약 59 품목의 독성시험성적에 대하여 농약관리법과 GHS에 따라 피부자극성 및 안점막자극성을 산출 및 분류한 결과, 피부자극성 경도 및 중도로 분류된 품목에서 GHS 기준에 의해 '분류되지 않음'으로 구분된 품목이 전체의 25.5%였고 안점막자극성 경도에서 GHS 기준에 의해 '분류되지 않음'으로 구분된 품목이 45.8%로 나타나 자극성 구분이 완화되었다. 반면 비가역적 반응의 평가에 의해 경도와 중도에서 피부자극성이 피부부식성 구분 1로 보다 높게 평가된 품목이 6.8%, 안점막자극성이 심한 눈손상 구분 1로 보다 높게 평가 된 품목이 5.1%였다. 농약 품목의 시험성적을 이용한 GHS 분류와 품목에 함유된 원제의 자극성 구분과 함유량을 이용하여 GHS 혼합물 분류기준에 따라 분류한 결과를 비교한 결과, 피부자극성에서는 25.5% 품목의 자극성 구분이 완화되었고, 3.4% 품목의 자극성 구분이 강화되었다. 안점막자극성에서는 25.4%의 품목에 대한 자극성 구분이 완화되었고 11.9%의 품목의 자극성 구분이 강화되었다. 농약 품목에 GHS를 도입할 경우, 상이한 분류 결과를 보완할 수 있는 방안 마련이 필요하다고 사료된다.

심한 회전근 개 부전을 동반한 환자에 대한 역형 견관절 전치환술 - 최소 3년 추시 결과 - (Reverse Total Shoulder Arthroplasty in Patients with Severe Rotator Cuff-Deficient Shoulder - A Minimum Three-Year Follow-up Study -)

  • 김명선;여제형
    • Clinics in Shoulder and Elbow
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    • 제16권2호
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    • pp.73-83
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    • 2013
  • 목적: 심한 회전근 개 부전을 동반한 관절와 상완 관절염 환자나 가성 마비 환자들에 대한 치료로 역형 견관절 전치환술 시행한 후 최소 3년 추시 결과를 알아보고자 하였다. 대상 및 방법: 2007년 7월부터 2010년 7월까지 본원에서 역형 견관절 전치환술을 시행 받은 13명(남자: 3명, 여자: 10명)의 환자를 대상으로 하였다. 추시 기간은 평균 54.2개월(37~74개월)이었으며, 임상적 결과는 동통에 대한 VAS 점수, 능동적 관절 운동 범위, ASES 평가 방법, Korean 견관절 평가 방법을 이용하였으며, 수술 중 및 수술 후 합병증에 대해서 분석하였다. 결과: 총 13예 중 지연성 심부 감염을 제외한 11예의 동통에 대한 VAS 점수는 술 전 평균 7.5(6~10)점에서 술 후 1.5(0~4)점으로, 능동적 전방 거상은 $42.7(10{\sim}100)^{\circ}$ 에서 $129.1(110{\sim}180)^{\circ}$ 로, ASES 점수는 32.9(11.7~46.7)점에서 80.2(58.3~95.0)점으로, KSS 점수는 36.8(24~47)점에서 78.4(61~92)점으로 호전되었다. 합병증으로는 다양한 정도의 관절와 절흔이 총 13예 중 12예, 수술 중 관절와 골절 1예, 일시적인 신경 손상이 2예 발생하였고, 지연성 심부 감염이 2예 발생하여 재수술이 필요하였다. 결론: 역형 견관절 전치환술은 최소 3년의 추시 결과, 심한 회전근 개 부전을 동반한 관절와 상완 관절염 환자나 가성 마비 환자들에 대해 효과적인 치료 방법의 하나로 생각된다. 그러나, 술 후 재수술이 필요한 지연성 심부 감염 등의 심각한 합병증과 수술 술기와 관련한 합병증의 발생 가능성을 고려해 볼 때 숙련된 의사에 의해 매우 신중하게 시행 되어져야 할 것으로 사료된다.