• Title/Summary/Keyword: 재조합

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Biochemical Analysis of Baculovirus-insect Cell Interaction: I. Improved Recombinant ${\beta}-Galactosidase$ Production Using Medium Additives at AcNPV Infection of Insect Cells (Baculovirus-곤충세포 상호반응에 대한 생화학적 연구 -I. AcNPV의 곤충세포 감염시 배지 첨가물을 이용한 재조합 ${\beta}-galactosidase$ 생산 향상-)

  • Lee, Ki-Woong;Kim, Tae-Yong;Chung, In-Sik
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.485-489
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    • 1995
  • The medium additives such as fatty acid, lipid, mannose, folic acid, $CaCl_2$ were examined to enhance recombinant ${\beta}-galactosidase\;({\beta}-gal)$ production in T-flask and air-lift bioreactor. The addition of each component. such as cholesterol, tocopherol, tricaprylin, mannose and folic acid at AcNPV infection of Tn5B1-4 cells enhanced ${\beta}-gal$ production, whereas the addition of $CaCl_2$ did not increase ${\beta}-gal$ production. The recombinant ${\beta}-gal$ production using the infection medium supplemented with a mixture of 0.34 mM cholesterol, 2.2 mM mannose and 0.045 M folic acid was enhanced 2 fold in an air-lift bioreactor, compared to the basal medium.

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Enantioselective Hydrolysis for Preparing (S)-Styrene Oxide in Organic Solvents Using Recombinant Escherichia coli Expressing Protein-engineered Epoxide Hydrolase of Mugil cephalus (Mugil cephalus 유래 에폭사이드 가수분해효소를 발현하는 재조합 대장균을 이용한 유기용매에서의 (S)-Styrene Oxide 제조를 위한 입체선택적 가수분해 반응)

  • Lee, Ok Kyung;Lee, Eun Yeol
    • Applied Chemistry for Engineering
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    • v.23 no.6
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    • pp.599-603
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    • 2012
  • The enantioselective hydrolysis of racemic styrene oxide in organic solvents was conducted using a recombinant E. coli expressing protein-engineered Mugil cephalus epoxide hydrolase (McEH). The volumetric total activity of the recombinant E. coli was enhanced 2.2-fold by IPTG induction at a mid-exponential growth phase. Among organic solvents with different log P values, isooctane was chosen based on the high activity and the enantioselectivity of McEH. Some lyoprotectants such as skim milk or sucrose enhanced the McEH activity. Enantiopure (S)-Styrene oxide with a 98% ee was obtained from the racemic styrene oxide with a 53.6% yield based on its theoretical yield in isooctane.

포장과 법률 - 자원의 절약과 재활용 촉진에 관한 법률 일부개정안

  • (사)한국포장협회
    • The monthly packaging world
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    • s.221
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    • pp.116-129
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    • 2011
  • 환경부는 생산자책임재활용제도(EPR)의 성공적 운영을 위해 의무생산자의 재활용의무이행방법(직접, 개별위탁, 공제조합)을 개선하고 재활용 이행에 있어 회수 의무를 분명히 하며 재활용사업공제조합을 복수화하되 소규모 조합 난립을 막기 위해 조합 인가 및 취소 요건을 규정하고 포장재 분야 공제조합은 통합하는 등 조합 설립형태를 합리적으로 개선하고 재활용지원금 차등지원 근거 마련 등 조합 운영방식을 효율적으로 개선하려는 취지 아래 자원의 절약과 재활용 촉진에 관한 법률을 일부 개정했다. 이번 개정안에서는 또한 그 밖에 현행 제도 운영과정에서 나타난 미비점을 개선 보완하고자 했다. 본 고에서는 주요 내용 및 변경된 법안을 살펴보도록 한다.

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포장과 법률 - 자원의 절약과 재활용 촉진에 관한 법률 일부개정안

  • (사)한국포장협회
    • The monthly packaging world
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    • s.222
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    • pp.110-121
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    • 2011
  • 환경부는 생산자책임재활용제도(EPR)의 성공적 운영을 위해 의무생산자의 재활용의무이행방법(직접, 개별위탁, 공제조합)을 개선하고 재활용 이행에 있어 회수 의무를 분명히 하며 재활용사업공제조합을 복수화하되 소규모 조합 난립을 막기 위해 조합 인가 및 취소 요건을 규정하고 포장재 분야 공제조합은 통합하는 등 조합 설립형태를 합리적으로 개선하고 재활용지원금 차등지원 근거 마련 등 조합 운영방식을 효율적으로 개선하려는 취지 아래 자원의 절약과 재활용 촉진에 관한 법률을 일부 개정했다. 이번 개정안에서는 또한 그 밖에 현행 제도 운영과정에서 나타난 미비점을 개선 보완하고자 했다. 본 고에서는 주요 내용 및 변경된 법안을 살펴보도록 한다.

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재조합 효모를 이용한 항혈전 단백질 히루딘 발효 생산공정의 최적화

  • Kim, Myeong-Dong;Gang, Hyeon-A;Lee, Sang-Gi;Seo, Jin-Ho
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.99-102
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    • 2001
  • Recombinant Saccharomyces cerevisiae strains harboring various copy numbers of hirudin gene were developed to study dependency of hirudin expression level on its gene copy number. A linear relationship between the copy number of hirudin expression cassette and hirudin expression level was observed up to 10 copies. A double <5-integration vector truncated wi 디 1 the unnecessary bacterial genes before yeast transformation showed a four-fold increase in transformation efficiency and a 1.3-fold enhancement in hirudin expression level compared with a single <5 system. Gratuitous hirudin expression strain was developed by disrupting the GALl gene of S. cerevisiae. Glucose that was fed in a limited manner effectively supported cell growth and hi겨din expression by the gratuitous strain. Effects of methanol concentrations on hirudin production in recombinant Hansenula polymorpha were investigated in continuous and fed-batch cultures. At a steady-state of continuous culture, an optimum methanol concentration of 1.7 g/L was determined at a dilution rate of 0.18 $h^{-1}$ with 1.8 mg/L ${\cdot}$ h hirudin productivity.

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Development of Recombinant Saccharomyces cerevisiae Expressing Epoxide Hydrolase for the Preparation of Chiral Phenyl Oxirane (광학활성 Phenyl Oxirane 제조용 유전자 재조합 생촉매 개발)

  • 이수정;이은정;김초희;이지원;김희숙;이은열
    • Journal of Life Science
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    • v.13 no.1
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    • pp.105-109
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    • 2003
  • Epoxide hydrolase (EH) gene from Aspergillus niger #33 was cloned from cDNA library generated by reverse transcriptase-polymerase chain reaction (RT-PCR). The nucleotide sequence analysis revealed that the deduced amino acid sequence was almost similiar to that of A. niger LCP521 previously reported. The cloned EH gene was transformed into Saccharomyces cerevisiae and expressed by addition of galactose. The recombinant S. cerevisiae showed hydrolytic activity toward racemic phenyl oxirane substrate based on chiral GC analysis, and can be used as a potential biocatalyst for the preparation of chiral phenyl oxirane.

Enhanced Expression of Foreign Gene in Baculovirus-Infected Insect Cells Using a Silkworm Hemolymph (누에 체액을 이용한 외래 유전자의 발현효율 증대)

  • 우수동;김혜성
    • Journal of Sericultural and Entomological Science
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    • v.37 no.2
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    • pp.181-185
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    • 1995
  • To enhance expression of foreign gene by the novel expression vector, pBmKSK1, of Bombyx mori nuclear polyhedrosis virus, E. coli $\beta$-galactosidase gene expressing recombinant virus was infected in BmN-4 cells and various concentrations of silkworm hemolymph were added to the recombinant virus-infected BmN-4 cells containing fetal bovine serum. The expression efficiency of foreign gene was determined by $\beta$-galactosidase activity in the culture media. The results showed that the silkworm hemolymph was effective to expression of foreign gene in the BmN-4 cells, suggesting that the silkworm hemolymph could be substituted for fetal bovine serum in the BmN-4 cells to enhance expression of foreign gene.

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Characterization of Recombinant Derivatives of pJY711 of Multicopy Streptomyces Plasmid (Multicopy Streptomyces 플라스미드 pJY711의 재조합 유도체의 특성)

  • 염도영;공인수;유주현
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.35-40
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    • 1990
  • The restriction clevage map of multi-copy recombinant plasmid, pJY712(8.1kb), carrying the thiostrepton resistance gene(tsr) was determined. pJY712 had a broad host range in Streptomyces and contained single BglII site for cloning purpose. The plasmid showed the phenomenon of lethal zygosis ($Ltz^{+}$). Transformation frequency of pJY712 was $5.0\times 10^{4}$ transformants per ug plasmid DNA (TFU) in S. lividans. Plasmid pJY713 was constructed by inserting the tyrosinase gene(mel) into the BclI site of pJY712. Recombinant plasmid pJY714 carrying the mel gene was constructed by in vitro deletion of a segment (1.9kb BglII-BclI fragment) from pJY713.

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Molecular Cloning and Nucleotide Sequence of Endo-Inulinase Gene from Xanthomonas oryzae #5

  • Kim, Byeong-U;Kim, Mi-Rang;Yu, Dong-Ju
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.655-659
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    • 2000
  • A 11.5-kb DNA fragment containing an endo-inulinase gene was cloned from Xanthomonas oryzae #5. It contained a single open reading frame of 3,999bp, encoding a polypeptide composed of signal peptide of 32 amino acids and mature protein of 1,301 amino acids. From the comparison of amino acids sequences with fructan hydrolases, inulinase, levanase and CFTase, the sequence of the endo-inulinase had highly homology of 72% with CFTase of B. circulans, and six highly conserved regions including the ${\beta}-fructosidase$ motif were found.

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Optimization of Environmental Conditions for Hirudin Production from Recombinant Saccharomyces cerevisiae (재조합 효모를 이용한 Hirudin 발효생산조건의 최적화)

  • 이동훈;서진호
    • KSBB Journal
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    • v.9 no.1
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    • pp.8-15
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    • 1994
  • The research has been carried out to optimize a recombinant S. cerevisine fermentation process for the production of an anticoagulant hirudin. The structural gene coding for hirudin was combined with the GAL10 promoter for controlled expression, the MFal signal sequence for hirudin secretion, and the GAL7 terminator for transcriptional termination. Growth medium composition and environmental conditions were optimized for maximizing cell growth and final hirudin concentration. The optimized conditions included yeast extract 40g/$\ell$, casamino acid 5g/$\ell$, g1ucose 20g/$\ell$, galactose 30g/$\ell$, DO 50% and temperature $30^{\circ}C$. These conditions yielded the specific cell growth rate of $0.13hr^{-1}$, the final cell density of 30g cell/$\ell$ and the final hirudin concentration of 64mg/$\ell$ in the batch fermentation with a 2.5$\ell$ jar fermentor.

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