Polyphenol oxidase(PPO) isolated from the crude extract of ascidian, Halocynthia roretzi, showed higher affinity for catechol than tyrosine or DL-DOPA. Successful enzyme assay could be performed at $25^{\circ}C$, 10min. by mixing 0.2ml of crude enzyme extract with 2.8ml of 0.13M catechol in 0.1M sodium phosphate buffer(pH 6.4). The specific activity of PPO which had been purified with a combination of ammonium sulfate treatment, ion exchange chromatography on DEAE-cellulose, and gel filtration on Sepharose 6B was 13-fold disc gel electrophoresis. The activity of PPO was stable from pH 5.0 to 8.0 and showed the peak activity at pH 6.4 .The optimum reaction temperature for PPO oxidation on catechol was 35$^{\circ}C$ and those enzyme were heat stable up to 4$0^{\circ}C$. Molecular weigth of the enzyme was estimated about 170kDa. One molecule was found to be composed of gour subunits. Two of them had molecular weigh of 55kDa and the others 30kDa. The {TEX}$K_{m}${/TEX} values, {TEX}$V_{max}${/TEX} and catalytic efficiency({TEX}$V_{max}${/TEX}/{TEX}$K_{m}${/TEX}) for catechol were 0.12mM, 2.5mM/liter/min. and {TEX}$0.18min^{-1}${/TEX} respectively. The substrate affinity and electrophorectic pattern suggested that the enzyme of ascidian was considered to be not tyosine but catechol oxidase.
Thakur, Shikha;Sharma, Nirmal Kant;Thakur, Neerja;Bhalla, Tek Chand
Microbiology and Biotechnology Letters
/
v.47
no.2
/
pp.259-268
/
2019
In this study, the extracellular metalloprotease from Serratia marcescens PPB-26 was purified to homogeneity via ethanol fractionation and DEAE-cellulose column chromatography. Thus, a 3.8-fold purification was achieved with a 20% yield and specific activity of 76.2 U/mg. The purified protease was a 50-kDa monomer whose optimum pH and temperature for activity were 7.5 and $30^{\circ}C$ respectively; however, it was found to remain active in the 5-9 pH range and up to $40^{\circ}C$ for 6 h. The protease had a half-life of 15 days at $4^{\circ}C$, an optimum reaction time of 10 min, and an optimum substrate (casein) concentration of 0.25%. Furthermore, the Michaelis constant ($K_m$) and reaction velocity ($V_{max}$) of the protease were calculated to be 0.28% and $111.11{\mu}moles/(min{\cdot}mg)^{-1}$, respectively. The protease was stable when subjected to metal ions (2 mM), showing increased activity with most (especially $CoCl_2$ and $MgSO_4$ (30.54% increase)). It was also stable when exposed to oxidizing agents, bleaching agents, and detergents (5% v/v for 60 min). It retained 93% of its activity in non-ionic detergents (Tween-20, Tween-80, and Triton X-100). Moreover, wash performance analysis in commercial detergents (Ariel and Tide) showed that not only was the protease capable of protein stain removal, but also reduced cleaning time by 80% when added to detergents. Thus, the Serratia marcescens PPB-26 metalloprotease appears to be a promising new candidate as a laundry additive in the detergent industry.
Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0~9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.
The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).
Liver microsomal epoxide hydrolase (mEH) is active in the detoxification of epoxide-containing reactive intermediate. Previous studies in this laboratory have shown that thiazole and pyrazine are efficacious inducers of mEH in rats with large increases in mEH mRNA levels (Carcinogensis, Kim et al, 1993). mEH was purified to electrophoretic homogeneity from thiazole-induced rat hepatic microsomes using DEAE-cellulose column chromatography whereas another protein $({\sim}43\;kDa)$ was co-purified with mEH from pyrazine-induced rat hepatic micrsomes (200 mg/kg body weight/day, ip, 3d). The antibody raised from a rabbit against mEH protein purified from thiazole-induced rat hepatic microsomes appeared to specifically recognize mEH protein in rat hepatic microsomes, as assessed by immunoblotting analysis. Immunoblotting analyses revealed a 10- and 7-fold increase in mEH levels in the hepatic microsomes isolated from thiazole- and pyrazine-treated rats, respectively. Moreover, immunoblotting analysis showed cross-reactivity of the mEH antibody with a 43 kDa protein in pyrazine-induced rat hepatic microsomes and with co-purified 43 kDa protein in purified fractions. The ratio between the 43 kDa protein and mEH in pyrazine-induced rat microsomes or in purified fractions was ${\sim}1$ to 15. N-terminal amino acid sequence analysis of both purified rat mEH and 43 kDa protein revealed that 10 out of 12 amino acids in N-terminus of the 43 kDa protein were identical with the mEH sequence with two amino acid residues of the 43 kDa protein undetermined. Either thiazole or pyrazine treatment, however, failed to increase the levels of mEH protein in rabbits while pyrazine caused elevation of the 43 kDa protein in this species, as determined by irnrnunoblotting analysis. These results demonstrated that treatment of rats with either thiazole or pyrazine causes elevation in hepatic mEH expiession whereas pyrazine treatment results in induction of another mEH-related 43 kDa protein and that a distinct species difference exists between rats and rabbits in the induction of mEH by these xenobiotics.
The fucoidan purified from Korean brown seaweed, Ecklonia stolonifera was characterized on molecular structure and blood anticoagulant activities. Extraction was conducted at $100^{\circ}C$ with water and repeated twice. The crude fucodian was 151.1g out of 20.0 kg of Ecklonia stolonifera. The Fucoidan-1, which was purified from crude fucoidan using calcium chloride and cetyl pyridium chloride (CPC), was 35.2% against crude fucoidan. Fucoidan-5 was obtained approximately 28.1% from Fucoidan-1 through DEAE-Toyopearl 650 M ion-exchange column chromatography and showed one band by cellulose acetate electrophoresis. The molecular weight of Fucoidan-5 was estimated to be about 21,000∼23,000 dalton by Sephacryl S-300 gel filtration chromatography. Fucoidan-5 consists of 35.7% of fucose and 4.3% of galactose and the molar ratio of fucose and sulfate was about one to one. IR spectrum of Fucoidan-5 showed absorption at $1240\;cm^{-1}\;and\;850\;cm^{-1}$ and specific rotation value, $[\alpha]$, was $[\alpha]$. These results suggests that the sulfate maybe bind at $C_{4}$ carbon on ${\alpha}-L-fucose$. Gas chromatograph of methyl alditol acetate revealed that Fucoidan-5 is a fucose containing sulfated polysaccharide with $({\alpha}l-2)\;or\;({\alpha}l-2)$ glycosidic linkage. Anti-thrombin activity of the Fucoidan-5 was estimated as 1.4 time stronger than heparin. From above results, the purification methods using CPC and ion exchange chromatography is effective tools for obtaining highly purified fucoidan from Gompi, Ecklonia stolonifera.
Park, Young-Il;Kim, Hee-Guen;Kim, Yoo-Young;Kim, In-Soo
Applied Biological Chemistry
/
v.39
no.6
/
pp.494-500
/
1996
Uptake of hen metal ions by water dropwort (Oenanthe stolonifera DC.) and its cadmium-binding protein were studied to probe for good method to remove heavy metal contaminants from environments. The plant was cultured in the culture medium (pH 7.0) containing the various concentrations of $Cd^{2+}$, $Cr^{3+}$ or $Pb^{2+}$, for 3 and 7 days. The residual heavy metals deposited in roots linearly increased as the metal ions concentration increased up to 17 ppm for $Cd^{2+}$, 20 ppm for $Cr^{3+}$ and 50 ppm for $Pb^{2+}$. Above these concentrations, the plant growth was inhibited and the uptake rates of the metal ions decreased. The heavy metals absorbed by the plant were mostly deposited in roots. In particular, the residual concentration of lead in roots was about four times higher than those of cadmium and chromium. When cultured in the medium containing 20 ppm of each metal ion, 80% of cadmium, 90% of cromium and 96% of lead were deposited in roots out of the total residual metal ions in the plant. These values correspond to 6.1 mg of cadmium, 5.2 mg of chromium and 23.6 mg of lead per one gram of roots tissue on a dry weight basis. A cadmium-binding protein was partially purified by extraction, gel filtration and DEAE-Cellulose chromatography from water dropworts that was grown in the medium containing 20 ppm $Cd^{2+}$. The purified protein was a single band on SDS- and non-denaturing- polyacrylamide gel electrophoresis. Its molecular mass was estimated to be ca. 5,000 dalton by gel filteration. Analysis of amino acid composition of the protein indicated that it had a typical amino acid composition of heavy metal-binding protein in that it contained 27% of acidic amino acids and 9.9% of cysteine. However, it is likely that the protein is a new plant metal-binding protein, since its amino acid composition is somewhat different from those of phytochelatins that have been known so far.
YANG Huyn-Pil;LEE An-Jong;KIM Yong-Tae;KIM Se-Kwon
Korean Journal of Fisheries and Aquatic Sciences
/
v.27
no.5
/
pp.482-494
/
1994
Most of carotenoprotein complexes have been extracted by using buffered solutions. However, in this study carotenoprotein from the muscle of Blue mussel(Mytilus edulis) was extracted by a detergent such as Triton X-100. It was purified and characterized by $20\%$ (w/v) $(NH_4)_2SO_4$, DEAE-cellulose ion exchange and Sephacryl S-300 gel filtration. The carotenoprotein(${\lambda}_{max}=462nm$) had an approximate M. W. of 372KDa(gel filtration). SDS-PAGE analysis of the carotenoprotein indicated the presence of four polypeptides of 60KDa($23.70\%$), 46.9KDa($9.14\%$), 26KDa($49.14\%$) and 13KDa($18.02\%$). Carotenoprotein denaturated by treatment with SDS to a final concentration of $0.2\%$ (w/v) caused a hypsochromic shift of ${\lambda}_{max}$ from 462nm to 456nm. The carotenoprotein contained lipids as structure units. The amino acid composition of the carotenoprotein contained large essential amino acid amounts of $62.8\%$, and the content of threonine($35.9\%$) was higher than other amino acids, but histidine, methionine and proline were not present. In the carotenoprotein, the major fatty acids were $C_{16:4},\;C_{16:0},\;C_{20:5}\;and\;C_{22:6}$. The percentages of polyunsaturated fatty acids($62.4\%$) were higher compared to other fatty acids(saturated fatty acids $19.6\%$, monounsaturated fatty acids $18.0\%$). Carotenoid was extracted from the carotenoprotein by acetone and it was separated into five different components by preparative TLC(benzene:petroleum ether:acetone=69:17:14). The major components of carotenoid were mytiloxanthin($74.79\%$) and 3,4,3'- trihydroxy-7',8'-didehydro-${\beta}$-carotene($18.26\%$), and they were at least presented as prosthetic groups of carotenoprotein.
Jo, Hang Soo;Choi, Doo Jin;Chung, Mi Ja;Park, Jae Kweon;Park, Yong Il
The Korean Journal of Mycology
/
v.40
no.4
/
pp.229-234
/
2012
A key molecule in the pathogenesis of Alzheimer's disease (AD) is the ${\beta}$-amyloid peptide ($A{\beta}$) generated by ${\beta}$-secretase activity, an aspartic protease. This study was designed to evaluate inhibitory effect of the high-molecular weight water-soluble polysaccharides (Et-P) isolated and purified from Phellinus linteus fruiting body on ${\beta}$-secretase activity. The Et-P was purified from the hot water extract of Phellinus linteus fruiting body mainly by 75% ethanol precipitation and DEAE-Cellulose column chromatography. From the DEAE-Cellulose chromato-gram and molecular weight analysis, the Et-P was shown to be a mixture of three polysaccharides with molecular mass of 1,629, 1,294, and 21 kDa, respectively. The monosaccharide composition of Et-P was determined to be glu-cose, galactose, and mannose as major sugars, glucose being the most prominent one (48% in mole percentage). The elemental analysis and FT-IR analysis suggested that Et-P is typical polysaccharides having at least partially ${\beta}$-linkages and possible existing as complex with phenolic compounds. The laminarinase digestion and HPAEC-PAD analysis suggested that Et-P is a variant of beta-(1,3)-glucans. The Et-P showed DPPH radical scavenging activity and, especially, a significant inhibitory activity on ${\beta}$-secreatase activity (48% inhibitin at 100 ${\mu}g/mL$), suggesting that they may inhibit the formation of $A{\beta}$ which is the major causative of Alzheimer's disease. The results of this study suggest that the water soluble polysaccharides of Phellinus linteus fruiting body can be a potent material for the development of preventive or therapeutic agents for AD.
As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.
본 웹사이트에 게시된 이메일 주소가 전자우편 수집 프로그램이나
그 밖의 기술적 장치를 이용하여 무단으로 수집되는 것을 거부하며,
이를 위반시 정보통신망법에 의해 형사 처벌됨을 유념하시기 바랍니다.
[게시일 2004년 10월 1일]
이용약관
제 1 장 총칙
제 1 조 (목적)
이 이용약관은 KoreaScience 홈페이지(이하 “당 사이트”)에서 제공하는 인터넷 서비스(이하 '서비스')의 가입조건 및 이용에 관한 제반 사항과 기타 필요한 사항을 구체적으로 규정함을 목적으로 합니다.
제 2 조 (용어의 정의)
① "이용자"라 함은 당 사이트에 접속하여 이 약관에 따라 당 사이트가 제공하는 서비스를 받는 회원 및 비회원을
말합니다.
② "회원"이라 함은 서비스를 이용하기 위하여 당 사이트에 개인정보를 제공하여 아이디(ID)와 비밀번호를 부여
받은 자를 말합니다.
③ "회원 아이디(ID)"라 함은 회원의 식별 및 서비스 이용을 위하여 자신이 선정한 문자 및 숫자의 조합을
말합니다.
④ "비밀번호(패스워드)"라 함은 회원이 자신의 비밀보호를 위하여 선정한 문자 및 숫자의 조합을 말합니다.
제 3 조 (이용약관의 효력 및 변경)
① 이 약관은 당 사이트에 게시하거나 기타의 방법으로 회원에게 공지함으로써 효력이 발생합니다.
② 당 사이트는 이 약관을 개정할 경우에 적용일자 및 개정사유를 명시하여 현행 약관과 함께 당 사이트의
초기화면에 그 적용일자 7일 이전부터 적용일자 전일까지 공지합니다. 다만, 회원에게 불리하게 약관내용을
변경하는 경우에는 최소한 30일 이상의 사전 유예기간을 두고 공지합니다. 이 경우 당 사이트는 개정 전
내용과 개정 후 내용을 명확하게 비교하여 이용자가 알기 쉽도록 표시합니다.
제 4 조(약관 외 준칙)
① 이 약관은 당 사이트가 제공하는 서비스에 관한 이용안내와 함께 적용됩니다.
② 이 약관에 명시되지 아니한 사항은 관계법령의 규정이 적용됩니다.
제 2 장 이용계약의 체결
제 5 조 (이용계약의 성립 등)
① 이용계약은 이용고객이 당 사이트가 정한 약관에 「동의합니다」를 선택하고, 당 사이트가 정한
온라인신청양식을 작성하여 서비스 이용을 신청한 후, 당 사이트가 이를 승낙함으로써 성립합니다.
② 제1항의 승낙은 당 사이트가 제공하는 과학기술정보검색, 맞춤정보, 서지정보 등 다른 서비스의 이용승낙을
포함합니다.
제 6 조 (회원가입)
서비스를 이용하고자 하는 고객은 당 사이트에서 정한 회원가입양식에 개인정보를 기재하여 가입을 하여야 합니다.
제 7 조 (개인정보의 보호 및 사용)
당 사이트는 관계법령이 정하는 바에 따라 회원 등록정보를 포함한 회원의 개인정보를 보호하기 위해 노력합니다. 회원 개인정보의 보호 및 사용에 대해서는 관련법령 및 당 사이트의 개인정보 보호정책이 적용됩니다.
제 8 조 (이용 신청의 승낙과 제한)
① 당 사이트는 제6조의 규정에 의한 이용신청고객에 대하여 서비스 이용을 승낙합니다.
② 당 사이트는 아래사항에 해당하는 경우에 대해서 승낙하지 아니 합니다.
- 이용계약 신청서의 내용을 허위로 기재한 경우
- 기타 규정한 제반사항을 위반하며 신청하는 경우
제 9 조 (회원 ID 부여 및 변경 등)
① 당 사이트는 이용고객에 대하여 약관에 정하는 바에 따라 자신이 선정한 회원 ID를 부여합니다.
② 회원 ID는 원칙적으로 변경이 불가하며 부득이한 사유로 인하여 변경 하고자 하는 경우에는 해당 ID를
해지하고 재가입해야 합니다.
③ 기타 회원 개인정보 관리 및 변경 등에 관한 사항은 서비스별 안내에 정하는 바에 의합니다.
제 3 장 계약 당사자의 의무
제 10 조 (KISTI의 의무)
① 당 사이트는 이용고객이 희망한 서비스 제공 개시일에 특별한 사정이 없는 한 서비스를 이용할 수 있도록
하여야 합니다.
② 당 사이트는 개인정보 보호를 위해 보안시스템을 구축하며 개인정보 보호정책을 공시하고 준수합니다.
③ 당 사이트는 회원으로부터 제기되는 의견이나 불만이 정당하다고 객관적으로 인정될 경우에는 적절한 절차를
거쳐 즉시 처리하여야 합니다. 다만, 즉시 처리가 곤란한 경우는 회원에게 그 사유와 처리일정을 통보하여야
합니다.
제 11 조 (회원의 의무)
① 이용자는 회원가입 신청 또는 회원정보 변경 시 실명으로 모든 사항을 사실에 근거하여 작성하여야 하며,
허위 또는 타인의 정보를 등록할 경우 일체의 권리를 주장할 수 없습니다.
② 당 사이트가 관계법령 및 개인정보 보호정책에 의거하여 그 책임을 지는 경우를 제외하고 회원에게 부여된
ID의 비밀번호 관리소홀, 부정사용에 의하여 발생하는 모든 결과에 대한 책임은 회원에게 있습니다.
③ 회원은 당 사이트 및 제 3자의 지적 재산권을 침해해서는 안 됩니다.
제 4 장 서비스의 이용
제 12 조 (서비스 이용 시간)
① 서비스 이용은 당 사이트의 업무상 또는 기술상 특별한 지장이 없는 한 연중무휴, 1일 24시간 운영을
원칙으로 합니다. 단, 당 사이트는 시스템 정기점검, 증설 및 교체를 위해 당 사이트가 정한 날이나 시간에
서비스를 일시 중단할 수 있으며, 예정되어 있는 작업으로 인한 서비스 일시중단은 당 사이트 홈페이지를
통해 사전에 공지합니다.
② 당 사이트는 서비스를 특정범위로 분할하여 각 범위별로 이용가능시간을 별도로 지정할 수 있습니다. 다만
이 경우 그 내용을 공지합니다.
제 13 조 (홈페이지 저작권)
① NDSL에서 제공하는 모든 저작물의 저작권은 원저작자에게 있으며, KISTI는 복제/배포/전송권을 확보하고
있습니다.
② NDSL에서 제공하는 콘텐츠를 상업적 및 기타 영리목적으로 복제/배포/전송할 경우 사전에 KISTI의 허락을
받아야 합니다.
③ NDSL에서 제공하는 콘텐츠를 보도, 비평, 교육, 연구 등을 위하여 정당한 범위 안에서 공정한 관행에
합치되게 인용할 수 있습니다.
④ NDSL에서 제공하는 콘텐츠를 무단 복제, 전송, 배포 기타 저작권법에 위반되는 방법으로 이용할 경우
저작권법 제136조에 따라 5년 이하의 징역 또는 5천만 원 이하의 벌금에 처해질 수 있습니다.
제 14 조 (유료서비스)
① 당 사이트 및 협력기관이 정한 유료서비스(원문복사 등)는 별도로 정해진 바에 따르며, 변경사항은 시행 전에
당 사이트 홈페이지를 통하여 회원에게 공지합니다.
② 유료서비스를 이용하려는 회원은 정해진 요금체계에 따라 요금을 납부해야 합니다.
제 5 장 계약 해지 및 이용 제한
제 15 조 (계약 해지)
회원이 이용계약을 해지하고자 하는 때에는 [가입해지] 메뉴를 이용해 직접 해지해야 합니다.
제 16 조 (서비스 이용제한)
① 당 사이트는 회원이 서비스 이용내용에 있어서 본 약관 제 11조 내용을 위반하거나, 다음 각 호에 해당하는
경우 서비스 이용을 제한할 수 있습니다.
- 2년 이상 서비스를 이용한 적이 없는 경우
- 기타 정상적인 서비스 운영에 방해가 될 경우
② 상기 이용제한 규정에 따라 서비스를 이용하는 회원에게 서비스 이용에 대하여 별도 공지 없이 서비스 이용의
일시정지, 이용계약 해지 할 수 있습니다.
제 17 조 (전자우편주소 수집 금지)
회원은 전자우편주소 추출기 등을 이용하여 전자우편주소를 수집 또는 제3자에게 제공할 수 없습니다.
제 6 장 손해배상 및 기타사항
제 18 조 (손해배상)
당 사이트는 무료로 제공되는 서비스와 관련하여 회원에게 어떠한 손해가 발생하더라도 당 사이트가 고의 또는 과실로 인한 손해발생을 제외하고는 이에 대하여 책임을 부담하지 아니합니다.
제 19 조 (관할 법원)
서비스 이용으로 발생한 분쟁에 대해 소송이 제기되는 경우 민사 소송법상의 관할 법원에 제기합니다.
[부 칙]
1. (시행일) 이 약관은 2016년 9월 5일부터 적용되며, 종전 약관은 본 약관으로 대체되며, 개정된 약관의 적용일 이전 가입자도 개정된 약관의 적용을 받습니다.