• Title/Summary/Keyword: DIGEST

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Biochemical Characterization of Exoribonuclease Encoded by SARS Coronavirus

  • Chen, Ping;Jiang, Miao;Hu, Tao;Liu, Qingzhen;Chen, Xiaojiang S.;Guo, Deyin
    • BMB Reports
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    • v.40 no.5
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    • pp.649-655
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    • 2007
  • The nsp14 protein is an exoribonuclease that is encoded by severe acute respiratory syndrome coronavirus (SARS-CoV). We have cloned and expressed the nsp14 protein in Escherichia coli, and characterized the nature and the role(s) of the metal ions in the reaction chemistry. The purified recombinant nsp14 protein digested a 5'-labeled RNA molecule, but failed to digest the RNA substrate that is modified with fluorescein group at the 3'-hydroxyl group, suggesting a 3'-to-5' exoribonuclease activity. The exoribonuclease activity requires $Mg^{2+}$ as a cofactor. Isothermal titration calorimetry (ITC) analysis indicated a two-metal binding mode for divalent cations by nsp14. Endogenous tryptophan fluorescence and circular dichroism (CD) spectra measurements showed that there was a structural change of nsp14 when binding with metal ions. We propose that the conformational change induced by metal ions may be a prerequisite for catalytic activity by correctly positioning the side chains of the residues located in the active site of the enzyme.

A Study on the Physiological Symptoms and Pathological Symptoms of Vietnamese according to Sasang Constitutions (사상체질에 따른 베트남인의 소증 및 병증 특성 연구)

  • Kwon, Ji-Hye;Park, Hye-Joo;Pham, Duc Duong;Dong, Sang-Oak;Jang, Eun-Su;Lee, Si-Woo;Kim, Sang-Hyuk;Yoo, Jong-Hyang
    • Journal of Sasang Constitutional Medicine
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    • v.25 no.4
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    • pp.297-305
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    • 2013
  • Objectives In this study, the Sasang Constitutional diagnosis were carried out to Vietnamese subject living in Vietnam, and the external applicability of Sasang Constitutional Medicine was evaluated by analyzing the physiological symptoms and the pathological symptoms according to their constitutions. Methods Total 170 Vietnamese subjects who live in Hanoi were invited from Sep. 2012 to Jun. 2013 for the research, and subjects were diagnosed by 2 Sasang Constitution Medicine specialists. The collected data were analyzed in terms of the physiological symptoms and the pathological symptoms according to the Sasang Constitutions. Results There were significantly different in the physiological symptoms according to Sasang Constitutions, which were 'digest', 'perspiration', 'excrement', 'hot and cold', 'water' and in the pathological symptoms, which were 'sting pain in head' in Soyangin, 'back pain' in Soeumin, 'chest discomfort' in Taeeumin, 'skin discomfort' in Taeeumin, 'fatigue in the afternoon' in Soyangin, 'digestion' in Soeumin. Conclusions The Sasang Constitution Medicine which is a Korean traditional medicine is also applicable to foreign countries, and it can contribute to the health promotion.

Adverse Event Study on Ojeok-san Extract Powders RCT in Low Back Pain Patients (요통환자 대상 오적산 엑스산 투여 임상시험 이상반응 연구)

  • Yoon, Eun-Hye;Jang, Min-Gee;Hwang, Ji-Hoo;Lee, Seung-Hoon;Jeong, Won-Je;Lee, Jae-Dong;Lee, Seung-Deok;Kim, Kyung-Ho;Kim, Kap-Sung;Kim, Eun-Jung
    • Journal of Acupuncture Research
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    • v.27 no.4
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    • pp.195-202
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    • 2010
  • Objectives : This study was performed to report adverse events in Ojeok-san extract powders RCT. Methods : 180 patients with LBP were randomized into groups for a double blind, placebo-controlled clinical study. They received Ojeok-san simple Extract, Ojeok-san complex Extract or placebo in powders, orally dissolved 3times a day for 4weeks. During 4weeks, we researched the symptoms and duration of adverse events and Digest ability, Milk hypersensitivity, Starch hypersensitivity, Caramel hypersensitivity, West-med hypersensitivity, Herb-med hypersensitivity in patients who had adverse events. Results : 52patients(28.9%) had adverse events and of them 44patients(28.9%) had lost adverse events naturally. But 5patients(2.8%) were withdrawn from the research because of the adverse events. Most of the symptoms of adverse event was Gastrointestinal symptom(88.3%) and adverse events frequency was not different from Ojeok-san simple Extract, Ojeok-san complex Extract and placebo. Conclusions : Ojeok-san extract powders had adverse events no different from the placebo, which shows that it is a safe drug.

Efficient Hardware Design of Hash Processor Supporting SHA-3 and SHAKE256 Algorithms (SHA-3과 SHAKE256 알고리듬을 지원하는 해쉬 프로세서의 하드웨어 설계)

  • Choi, Byeong-Yoon
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.21 no.6
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    • pp.1075-1082
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    • 2017
  • This paper describes a design of hash processor which can execute new hash algorithm, SHA-3 and extendable-output function (XOF), SHAKE-256. The processor that consists of padder block, round-core block and output block maximizes its performance by using the block-level pipelining scheme. The padder block formats the variable-length input data into multiple blocks and then round block generates SHA-3 message digest or SHAKE256 result for multiple blocks using on-the-fly round constant generator. The output block finally transfers the result to host processor. The hash processor that is implemented with Xilinx Virtex-5 FPGA can operate up to 220-MHz clock frequency. The estimated maximum throughput is 5.28 Gbps(giga bits per second) for SHA3-512. Because the processor supports both SHA-3 hash algorithm and SHAKE256 algorithm, it can be applicable to cryptographic areas such as data integrity, key generation and random number generation.

Optimization of Culture Conditions for the Production of Diphtheria Toxin (디프테리아 toxin 생산을 위한 발효조건 최적화)

  • Cho, Min;Ryu, Yeon-Woo
    • KSBB Journal
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    • v.14 no.2
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    • pp.241-247
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    • 1999
  • Experimental studies were carried out to optimize the culture conditions of Corynebacterium diphtheriae for the production of diphtheria toxin. A new media which does not contain any meat digest products was selected. The main ingredient of new medium was enzymatic digests of casein known as NZ-Case. In fermenter experiments, the toxin production was increased with the increase of cell growth. The optimum initial pH of media, air flow rate and agitation speed were 7.0, 0.22, vvm and 400 rpm, respectively. The contents of iron and calcium-phosphate precipitate were important for maximal cell growth and toxin production. The optimum concentration of iron was 0.3 mg/L and calcium-phosphate precipitate could serve in gradual supply of iron to maintain the optimal culture condition which is required for enhanced yield of toxin production. In potency test, the potency of toxoid from fermentor culture was higher than that from static culture. When diphtheria toxin is produced by fermentor culture, it is possible to produce higher levels of toxin and better toxoid quality in terms of safety, yield, productivity and immunity.

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Prevalence, Serotype Diversity, Genotype and Antibiotic Resistance of Listeria monocytogenes Isolated from Carcasses and Human in Korea

  • Oh, Hyemin;Kim, Sejeong;Lee, Soomin;Lee, Heeyoung;Ha, Jimyeong;Lee, Jeeyeon;Choi, Yukyung;Choi, Kyoung-Hee;Yoon, Yohan
    • Food Science of Animal Resources
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    • v.38 no.5
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    • pp.851-865
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    • 2018
  • This study investigated the prevalence of Listeria monocytogenes in slaughterhouses, and determined serovars and genotypes, and antibiotic resistance of the isolates obtained from slaughterhouses and humans in Korea. Two hundred ninety samples were collected from feces (n=136), carcasses [n=140 (cattle: n=61, swine: n=79)], and washing water (n=14) in nine slaughterhouses. Eleven human isolates were obtained from hospitals and the Korea Center for Disease Control and Prevention. Listeria monocytogenes was enriched and identified, using polymerase chain reaction (PCR) and 16S rRNA sequencing. Serovars and presence of virulence genes were determined, and genetic correlations among the isolates were evaluated by the restriction digest patterns of AscI. Antibiotic resistance of L. monocytogenes isolates were examined against 12 different antibiotics. Of 290 slaughterhouse samples, 15 (5.17%) carcass samples were L. monocytogenes positive. Most L. monocytogenes isolates possessed all the virulence genes, while polymorphisms in the actA gene were found between carcass and human isolates. Serovars 1/2a (33.3%) and 1/2b (46.7%) were the most frequent in carcass isolates. Genetic correlations among the isolates from carcass and clinical isolates were grouped within serotypes, but there were low geographical correlations. Most L. monocytogenes isolates were antibiotic resistant, and some strains showed resistance to more than four antibiotics. These results indicate that L. monocytogenes are isolated from carcass and human in Korea, and they showed high risk serotypes and antibiotic resistance. Therefore, intensive attentions are necessary to be aware for the risk of L. monocytogenes in Korea.

Anaerobic Biotreatment of Animal Manure - A review of current knowledge and direction for future research -

  • Hong, Jihyung
    • Journal of Animal Environmental Science
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    • v.11 no.2
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    • pp.97-102
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    • 2005
  • Anaerobic decomposition is one of the most common processes in nature and has been extensively used in waste and wastewater treatment for several centuries. New applications and system modifications continue to be adapted making the process either more effective, less expensive, or suited to the particular waste in question and the operation to which it is to be applied. Animal manure is a highly biodegradable organic material and will naturally undergo anaerobic fermentation, resulting in release of noxious odors, such as in manure storage pits. Depending on the presence or absence of oxygen in the manure, biological treatment process may be either aerobic or anaerobic. Under anaerobic conditions, bacteria carry on fermentative metabolisms to break down the complex organic substances into simpler organic acids and then convert them to ultimately formed methane and carbon dioxide. Anaerobic biological systems for animal manure treatment include anaerobic lagoons and anaerobic digesters. Methane and carbon dioxide are the principal end products of controlled anaerobic digestion. These two gases are collectively called biogas. The biogas contains $60\~70\%$ methane and can be used directly as a fuel for heating or electrical power generation. Trace amounts of ammonia and hydrogen sulfide ($100\~300\;ppm$) are always present in the biogas stream. Anaerobic lagoons have found widespread application in the treatment of animal manure because of their low initial costs, ease of operation and convenience of loading by gravity flow from the animal buildings. The main disadvantage is the release of odors from the open surfaces of the lagoons, especially during the spring warm-up or if the lagoons are overloaded. However, if the lagoons are covered and gases are collected, the odor problems can be solved and the methane collected can be used as a fuel. Anaerobic digesters are air-tight, enclosed vessels and are used to digest manure in a well-controlled environment, thus resulting in higher digestion rates and smaller space requirements than anaerobic lagoons. Anaerobic digesters are usually heated and mixed to maximize treatment efficiency and biogas production. The objective of this work was to review a current anaerobic biological treatment of animal manure for effective new technologies in the future.

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Key to the Korean Nematode-Trapping Fungi with Additional Descriptions of Arthrobotrys flagrans and A. superba (한국 선충 포식성곰팡이 분류검색표 및 Arthrobotrys flagrans 와 A. superba의 재기재)

  • Seo, Jongmin;Kang, Heonil;Kwon, Giyoon;Park, Namsook;Bae, Changhwan;Choi, Insoo
    • The Korean Journal of Mycology
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    • v.47 no.4
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    • pp.291-301
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    • 2019
  • Nematophagous fungi can capture, kill, and digest nematodes using a specific capturing organ. Of the nematophagous fungi, while Arthrobotrys flagrans and A. superba have been described previously, certain characteristics have not been described. For a detailed description of the two nematophagous fungi, the fungi were isolated from soil samples and produced in a pure culture. Morphological characteristics, such as predatory ability (according to the nematode species), shape, and size of predatory organ, conidia, and chlamydospore were investigated and they were used for identification of the fungal isolates along with molecular phylogenetic analysis. Furthermore, this study provides the classification key for 21 nematophagous species.

Purification, Characterization, and Partial Primary Sequence of a Major-Maltotriose-producing $\alpha$-Amylase, ScAmy43, from Sclerotinia sclerotiorum

  • Ben Abdelmalek-Khedher, Imen;Urdad, Maria Camino;Limam, Ferid;Schmitter, Jean Marie;Marzouki, M. Nejib;Bressollier, Philippe
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1555-1563
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    • 2008
  • A novel $\alpha$-amylase ($\alpha$-1,4-$\alpha$-D-glucan glucanohydrolase, E.C. 3.2.1.1), ScAmy43, was found in the culture medium of the phytopathogenic fungus Sclerotinia sclerotiorum grown on oats flour. Purified to homogeneity, ScAmy43 appeared as a 43 kDa monomeric enzyme, as estimated by SDS-PAGE and Superdex 75 gel filtration. The MALDI peptide mass fingerprint of ScAmy43 tryptic digest as well as internal sequence analyses indicate that the enzyme has an original primary structure when compared with other fungal a-amylases. However, the sequence of the 12 N-terminal residues is homologous with those of Aspergillus awamori and Aspergillus kawachii amylases, suggesting that the new enzyme belongs to the same GH13 glycosyl hydrolase family. Assayed with soluble starch as substrate, this enzyme displayed optimal activity at pH 4 and $55^{\circ}C$ with an apparent $K_m$ value of 1.66 mg/ml and $V_{max}$ of 0.1${\mu}mol$glucose $min^{-1}$ $ml^{-1}$. ScAmy43 activity was strongly inhibited by $Cu^{2+}$, $Mn^{2+}$, and $Ba^{2+}$, moderately by $Fe^{2+}$, and was only weakly affected by $Ca^{2+}$ addition. However, since EDTA and EGTA did not inhibit ScAmy43 activity, this enzyme is probably not a metalloprotein. DTT and $\beta$-mercaptoethanol strongly increased the enzyme activity. Starting with soluble starch as substrate, the end products were mainly maltotriose, suggesting for this enzyme an endo action.

NMR study of the interaction of T$_4$ Endonuclease V with DNA

  • 이봉진;유준석;임형미;임후강
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.267-267
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    • 1994
  • In order to obtain insight into the mechanism by which DNA containing a thymine photo-dimer is recognized by the excision repair enzyme, T$_4$ endonuclease V, we have taken NMR study of this protein and its complex with oligonucleotides. The conformations of five different DNA duplexes DNA I : d(GCGGATGGCG).d(CGCCTACCGC), DNA II d(GCGGTTGGCG) .d(CGCCAACCGC), DNA III : d(GCGGT ^ TGGCG) .d(CGCCAACCGC), DNA IV d(GCGGGCGGCG).d(CGCCCGCCGC) and DNA V d(GCGGCCGGCG) . d(CGCCGGCCGC) were studied by $^1$H NMR. The NMR spectra of these five DNA duplexes in the absence of the enzyme clearly show that the formation of a thymine dimer within the DNA induces only a minor distortion in the structure, and that the overall structure of B type DNA is retained. The photo-dimer formation is found to cause a large change in chemical shifts at the GC7 base pair, which is located at the 3'-side of the thymine dimer, accompanied by the major conformational change at the thymine dimer site. The binding of a mutant T$_4$ endonuclease V (E23Q), which is unable to digest DNA containing a thymine dimer, to the DNA duplex d(GCGGT ^ TGGCG)ㆍd(CGCCAACCGC) causes a large down-field shift in the imino proton resonance of GC7. Therefore, this position is thought to be either the crucial point of the interaction wi th T$_4$ endonuclease V, or the si to of a conformational change in the DNA caused by the binding of T$_4$ endonuclease V. Usually, it is very difficult to assign NMR peaks in DNA * protein complex because of severe peak overlaps. In order to overcome these peak overlaps, we used a method of deuterium incorporation.

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