• Title/Summary/Keyword: 단백질계 배지

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수성이상계를 이용한 재조합 hGM-CSF의 in situ 분리

  • Pyo, Se-Hong;Myeong, Hyeon-Jong;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.321-324
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    • 2002
  • In situ recovery of recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) was performed in transgenic Nicotiana tabacum cell suspension cultures. Aqueous two-phase systems (ATPS) were used to utilize its biocompatibility. Transgenic plant cells could be grown up to 15.7 g/L in normal medium and 18.6 g/L in ATPS composed of 6% (w/w) polyethylene glycol (PEG) 20,000 and 10% (w/w) dextran 2,000,000. It was proved that the A TPS was not harmful to cell growth. In addition, It is expected to recover hGM-CSF simultaneously with cell growth. Using this method, maximum hGM-CSF concentration at 1.64 ng/mL was obtained on day 3.

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Analysis and Conservation of Metal Thread Made of Proteinaceous Substrate - Golden Decorative Rank Badge of an Official Uniform Excavated from Baekryeong Im's Tomb in the 16th Century of Korea - (단백질계 배지로 이루어진 금속사의 분석과 보존처리 - 16세기 임백령 묘 출토 단령의 직금 흉배를 중심으로 -)

  • Noh, Soo-Jung;Oh, Joon-Suk
    • Journal of the Korean Society of Costume
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    • v.58 no.9
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    • pp.129-141
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    • 2008
  • Jikgeum(woven with supplementary golden wefts) hyungbae(rank badge) of danryung(official uniform) excavated from Im Backryung'tomb($1498{\sim}1546$) of the Joseon dynasty($1392{\sim}1910$) at Goyang, Gyunggi-Do in 2007, was in a critical condition because of serious collapse of substrate in metal thread. For conservation of hyungbae, metal thread was examined by different scientific methods(Light Microscope, Scanning Electron Microscope and Energy Dispersive X-Rray Spectrometry(SEM-EDS), Fourier Transform Infrared Spectroscopy(FT-IR)). Analytical data showed that metal thread was gilt membrane strip composed of gold leaves and proteinaceous substrate which was probably parchment. To protect collapse of substrate, 1% solution of Paraloid B-72 was infilterated into substrate for consolidation of substrate and it was adhered to warp of fabric in hyungbae, before wet cleaning. After wet cleaning, the most of the gold leaves were restored, which was confirmed by both the examination with the naked eye and the microscopic examination.

Heavy-Metal Adsorption by Recombinant Saccharomyces cerevisiae Harboring Multiple Copies of the CUP1 Gene (구리흡착 단백질 유전자를 함유하는 재조합 효모의 중금속 흡착)

  • 서진호;박상옥;김명동;한기철;전영석;안장우;한남수
    • KSBB Journal
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    • v.17 no.1
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    • pp.38-43
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    • 2002
  • Characteristics of cell growth and heavymetal adsorption by recombinant Saccharomyus cerevisiae strains harboring multiple copies of the CUP1 gene encoding metallothione (MT) protein were studied in batch cultures. Recombinant S. cerevisiae strains harboring multiple copies of the CUP1 gene were superior to the host and wild-type yeast strains in terms of cell growth and heavy metal removal, indicating that the copy number of the CUP1 gene for MT expression played an important role in the adsorption of heavy metals. It was suggested that the CUP1 promoter for the MT expression is induced by manganese and zinc as well as copper An optimum copper concentration for MT expression and concomitant adsorption of heavy metals by recombinant S. cerevisiae was found to be 0.31 mM. A nonionic surfactant Triton X-100 enhanced cell growth by 17.7% and removal of zinc by 6.1% compared with the control case.

Double-Stranded RNA-Dependent Protein Kinase Gene Expression in Tobacco Plant (연초식물체에서의 dsRNA 의존성 인산화 효소 유전자 발현)

  • 이청호;박희성
    • Korean Journal Plant Pathology
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    • v.11 no.2
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    • pp.173-178
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    • 1995
  • 동물계에서 항바이러스와관련된 dsRNA 의존성 인산화 효소(PKR)의 유전자를 식물체에서 발현시킬 경우 PKR에 의한 단백질합성 및 식물바이러스의 증식조절 가능성에 대한 기초자료를 확보하기 위하여 사람에서 분리된 PKR cDNA를 Agrobacterium 방법에 의하여 연초식물체(Nicotiana tabacum cv. Xanthi-nc)로 형질전환시켰다. HindIII/PstI처리에 의해 얻어지는 약 1.8kb의 phPKR cDNA절편을 일련의 유전자 조작 방법을 통하여 식물발현벡터인 pBI121에 도입하여, p12168을 재조합하였다. 이를 A. tumefaciens LBA 4404에 형질전환시켜 연초식물체형질 전환에 이용하였다. 2mg/l BA와 0.5mg/l NAA가 포함되고 100$\mu\textrm{g}$/ml의 kanamycin이 첨가된 MS배지에서 shooting시킨 후 phytohormone이 첨가되지 않은 MS배지상에서 rooting을 시켜 형질전환 연초식물체를 얻었으며, 형질전환식물체는 정상식물체와 유사한 생육양상을 나타내었다. 형질전환식물체의 유전자도입은 hPKR cDNA의 전사부여는 RT-PCR 방법에 의하여 확인되었다.

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Screening of Outstanding White Rot Fungi for Biodegradation of Organosolv Lignin by Decolorization of Remazol Brilliant Blue R and Ligninolytic Enzymes Systems (Remazol Brilliant Blue R 탈색능과 리그닌 분해 효소시스템을 이용한 유기용매 리그닌 생분해 우수 균주 선별)

  • Hong, Chang-Young;Kim, Ho-Yong;Jang, Soo-Kyeong;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.41 no.1
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    • pp.19-32
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    • 2013
  • In this study, outstanding white rot fungi for biodegradation of organosolv lignin were selected on the basis of their ligninolytic enzyme system. Fifteen white rot fungi were evaluated for their ability to decolorize Remazol Brilliant Blue R (RBBR) in SSC and MEB medium, respectively. Six white rot fungi (Ceriporiopsis subvermispora, Ceriporia lacerate, Fomitopsis insularis, Phanerochaete chrysosporium, Polyporus brumalis, and Stereum hirsutum) decolorized RBBR rapidly in SSC medium within 3 days. The protein contents as well as the activities of manganese peroxidase (MnP) and laccase for 6 selected fungi were determined on the SSC medium with and without organosolv lignin. Interestingly, extracellular protein concentrations were determined to relative higher for S. hirsutum and P. chrysosporium in the presence of organosolv lignin than others. On the other hands, each fungus showed a different ligninolytic enzyme pattern. Among them, F. insularis resulted the highest ligninolytic enzyme activities on incubation day 6, indicating of 1,545 U/mg of MnP activity and 1,259 U/mg of laccase activity. In conclusion, $STH^*$ and FOI were considered as outstanding fungi for biodegradation of organosolv lignin, because $STH^*$ showed high extracellular protein contents and ligninolytic enzyme activities over all, and ligninolytic enzyme activities of FOI were the highest among white rot fungi used in this study.

Microbial Extracellular Enzyme Detection on Agar Plates by Means of Fluorogenic Methylumbelliferyl-Substrates (Methylumbelliferyl 형광기질을 이용한 평판배지상의 미생물 체외 세포효소측정방법)

  • ;Hoppe, H.-G.
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.229-235
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    • 1990
  • A rapid and sensitive method to detect the extracellular enzymatic activity of bacteria colonies grown on agar plates is described. Selective agar media supplemented with protein, starch, chitin, Tween-80, etc. are conventionally used to detect biochemical properties of bacteria. It has been experimentally demonstrated with bacteria pure cultures that fluorogenic Methylumbelliferyl (MUF)-substrates are excellent substrate analogues for normally occurring polymers. Based on MUF-substrate hydrolysis the new method provides reliable qualitative estimates of extracellular enzymatic properties of bacteria within minutes using pure cultures as well as agar p;ates prepared for colony counts. Extracellular enzyme activities of heterotrophic bacteria from freshwater ecosystems and marine sediment using this method are discussed.

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Recovery of Intracellular Enzyme Using Phage Lysogen and Expanded Bed Adsorption (Phage Lysogen과 Expanded Bed Adsorption을 이용한 세포내 효소의 회수)

  • 장우진;윤현식구윤모
    • KSBB Journal
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    • v.11 no.6
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    • pp.663-668
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    • 1996
  • In the production of recombinant protein using E. coli, phage lysogen system can be usefully applied for simultaneously achieving protein production at high cell concentration and recovery by cell disruption in the same bioreactor. A major drawback of this system is that the intracellular product and complex broth components are mixed together in culture broth and hence purification efficiency is reduced. With the E. coli double-lysogen system, the expanded bed adsorption is very useful because the pretreatment processes in a routine bioseparation process can be done in a single column operation, and therefore may contribute towards lowering the operating cost of overall recovery/purification process. In the operation of EBA, it has been observed that the change in broth feed volume does not influence much the protein recovery in a tested range. The amount of protein adsorption per mL of resin was increased from $3.44{\times}106unit to 5.28{\times}106unit$ by doubling the column length. By two-fold increase of the column diameter, the ratio of protein concentration in eluent to that in feed was increased from 0.8 to 2.1. It is concluded from the present investigation that the increase of column length and diameter is necessary to enhance the protein adsorption amount per volume of resin and protein concentration in the eluent. The development of resins with various physical properties will be necessary for more extensive application of EBA.

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High-Level Expression of Aspergillus ficuum Acetyl Xylan Esterase Gene in Pichia pastoris, (Pichia pastoris에서 Aspergillus ficuum 유래 Acetyl Xylan Esterase 유전자의 과발현)

  • 임재명;김성구;박승문;남수완
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.305-311
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    • 2002
  • Acetyl xylan esterase gene (AXE) from Aspergillus ficuum was cloned and its Pichia expression plasmid, pPICZ$\alpha$C-AXE (4.6 kb), was constructed, in which the AXE gene was under the control of the AOXI promoter and connected downstream of mating factor u-1 signal sequence. The plasmid linearized by Sacl was integrated into the 5'AOXI region of the chromosomal DNA of P. pastoris. In the flask batch culture of P. pastoris transformant on methanol medium, the cell concentration and total AXEase activity reached at 6.0 g-dry cell weight/1 and 77 unit/ml after 36 h cultivation, respectively. In the fed-batch culture employing the optimized methanol and histidine feeding strategy, the cell concentration and total AXEase activity were significantly increased to about 97 g-dry cell weight/1 and 930 unit/ml. Most of AXEase activity (>90%) was found in the extracellular medium and the majority of extracellular protein (>80%) was AXEase enzyme (33.5 kDa). This result means that about 9.8 g/1 of AXEase protein was produced in the extracellular medium.

A Scientific Analysis of Gold Threads Used in Donggungbi-Wonsam(Ceremonial Robe Worn by a Crown Princess, National Folklore Cultural Heritage No.48) (동궁비 원삼에 사용된 금사의 과학적 분석)

  • Lee, Jang-Jon;An, Boyeon;Han, Kiok;Lee, Ryangmi;Yoo, Ji Hyun;Yu, Ji A
    • Journal of Conservation Science
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    • v.37 no.5
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    • pp.525-535
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    • 2021
  • This study identified material properties through scientific analysis on Jikgeumdan(satin with gold threads) from Donggungbi-Wonsam and the gold threads used in the embroidery. The Donggungbi-Wonsam's base of gold threads were estimated to have used mulberry fiber's Korean paper(Hanji) because non-wood-based fibers were observed. The X-ray spectrometer showed that the Tongsuseulan of Donggungbi-Wonsam was a flat gold thread of pure gold and Jikgeumdan of flat silver thread of its Saekdong and Hansam. High sulfur levels were detected in the flat silver thread, which appeared to have formed silver sulfide by either manufacturing process using sulfur or conservation environment. he dragon insignia's embroidery is also described as two types twisted gold threads; pure gold and alloying-gold and silver. while dragon insignia's border line is decorated with a twisted gold thread of pure gold. In particular, it was investigated that adhesives such as an animal glue, a protein-based compound by gas chromatography mass spectrometry. Additionally, XRF and Raman spectroscopy analysis on the mixture substances between the metal surface and the base paper of gold threads identified talc and quartz in the gold threads and Seokganju(hematite) in the flat silver threads.

Effect of N-sources and NaCl Concentrations in Media on the Intra-and Extracellular Proteinase Activities of Streptococcus cremoris $ML_4$ and Streptococcus lactis $ML_8$ (Streptococcus cremoris $ML_4$ 및 Streptococcus lactis $ML_8$의 생육중 질소원과 염농도가 세포내 및 세포외 프로테이나제 역가에 미치는 영향)

  • Chang, Hae-Choon;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.21 no.1
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    • pp.86-91
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    • 1989
  • To investigate the effect of NaCl concentration and nitrogen sources in the medium for the Streptococci on the intra- and extracellular proteinase(ICP and ECP) which have been known as one of the major causes for the bitter peptide formation, Streptococcus lactis $ML_8$ and Streptococcus cremoris $ML_4$ strains were incubated at 0-4% NaCl in the medium and Na-caseinate as a nitrogen source 0-100%, and the cell growth, ICP and ECP activities were analyzed. As the concentration of the NaCl in the medium increased, the growth and ECP activity decreased but the ICP $activity/10^{10}$ cells increased on the contrary. This implied that the NaCl in the medium affects only the ECP which is associated mainly to the cell wall and cell membrane but not the ICP activity. When the content of the caseinate instead of other low molecular nitrogen sources were increased in the medium, the cell growth was lowered while the ECP activities increased probably by induction of proteinase production.

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