• 제목/요약/키워드: Functional characterization

검색결과 798건 처리시간 0.031초

Achieving High Yield of Lactic Acid for Antimicrobial Characterization in Cephalosporin-Resistant Lactobacillus by the Co-Expression of theosphofructokinase and Glucokinase

  • Gong, Yahui;Li, Tiyuan;Li, Shiyu;Jiang, Zhenyou;Yang, Yan;Huang, Junli;Liu, Zhaobing;Sun, Hanxiao
    • Journal of Microbiology and Biotechnology
    • /
    • 제26권6호
    • /
    • pp.1148-1161
    • /
    • 2016
  • Lactobacilli are universally recognized as probiotics that are widely used in the adjuvant treatment of inflammatory diseases, such as vaginitis and enteritis. With the overuse of antibiotics in recent years, the lactobacilli in the human body are killed, which could disrupt the microecological balance in the human body and affect health adversely. In this work, cephalosporin-resistant Lactobacillus casei RL20 was obtained successfully from the feces of healthy volunteers, which possessed a stable genetic set. However, the shortage of lactic acid (72.0 g/l at 48 h) by fermentation did not meet the requirement for its use in medicine. To increase the production of lactic acid, the functional genes pfk and glk were introduced into the wild strain. A yield of 144.2 g/l lactic acid was obtained in the transgenic L. casei RL20-2 after fermentation for 48 h in 1 L of basic fermentation medium with an initial glucose concentration of 100 g/l and increasing antibacterial activity. These data suggested that L. casei RL20-2 that exhibited a high yield of lactic acid may be a potential probiotic to inhibit the spread of bacterial infectious diseases and may be used for vaginitis therapy.

A Human Fungal Pathogen Cryptococcus neoformans Expresses Three Distinct Iron Permease Homologs

  • Han, Kyunghwan;Do, Eunsoo;Jung, Won Hee
    • Journal of Microbiology and Biotechnology
    • /
    • 제22권12호
    • /
    • pp.1644-1652
    • /
    • 2012
  • Iron plays a key role in host-pathogen interactions. Microbial pathogens require iron for survival and virulence, whereas mammalian hosts sequester and withhold iron as a means of nutritional immunity. We previously identified two paralogous genes, CFT1 and CFT2, which encode homologs of a fungal iron permease, Cft1 and Cft2, respectively, in the human fungal pathogen Cryptococcus neoformans. Cft1 was shown to play a role in the high-affinity reductive iron uptake system, and was required for transferrin utilization and full virulence in mammalian hosts. However, no role of Cft2 has been suggested yet. Here, we identified the third gene, CFT3, that produces an additional fungal iron permease homolog in C. neoformans, and we also generated the cft3 mutant for functional characterization. We aimed to reveal distinct functions of Cft1, Cft2 and Cft3 by analyzing phenotypes of the mutants lacking CFT1, CFT2 and CFT3, respectively. The endogenous promoter of CFT1, CFT2 and CFT3 was replaced with the inducible GAL7 promoter in the wild-type strain or in the cft1 mutant for gain-of-function analysis. Using these strains, we were able to find that CFT2 is required for growth in low-iron conditions in the absence of CFT1 and that overexpression of CFT2 compensates for deficiency of the cft1 mutant in iron uptake and various cellular stress conditions. However, unlike CFT2, no clear phenotypic characteristic of the cft3 mutant and the strain overexpressing CFT3 was observed. Overall, our data suggested a redundant role of Cft2 in the high-affinity iron uptake and stress responses in C. neoformans.

매립 연령에 따른 침출수중에 존재하는 휴믹물질의 분광학적 특성 분석 (Spectroscopic Characterization of Humic Substances Present in Landfill Leachates with Respect to Landfilling Age)

  • 강기훈;신현상;박희경
    • 대한환경공학회지
    • /
    • 제22권6호
    • /
    • pp.1113-1126
    • /
    • 2000
  • 매립지 침출수에 의한 지하수의 오염으로 생성되는 대수층내 오염대 내에서 휴믹물질이 다른 오염물질의 이동 및 변환에 미치는 영향을 평가하기 위한 기초연구로서 매립지의 매립년한에 따른 침출수중에 존재하는 휴믹물질을 추출하여 그 특성을 분석하였다. 추출된 휴믹물질에 대해 원소분석을 실시하였으며, 분광학적 분석으로서 자외선-가시광선, 적외선, 형광 분광분석과 $^1H$$^{13}C$ 핵자기공명 분석을 실시하였다. 침출수중에 존재하는 휴믹물질은 토양 및 수중으로 부터 추출된 휴믹물질에 비해 구조적으로는 지방족 화합물에 의한 특성이 방향족 화합물에 의한 특성에 비교하여 우세하였으며, 이로부터 침출수 휴믹물질의 휴믹화 정도가 상대적으로 낮음을 알 수 있었다. 한편 매립지의 매립년한이 증가할수록 침출수 휴믹물질의 휴믹화 정도는 증가하는 특성을 보였다.

  • PDF

Thermal properties and mechanical properties of dielectric materials for thermal imprint lithography

  • Kwak, Jeon-Bok;Cho, Jae-Choon;Ra, Seung-Hyun
    • 한국전기전자재료학회:학술대회논문집
    • /
    • 한국전기전자재료학회 2006년도 하계학술대회 논문집 Vol.7
    • /
    • pp.242-242
    • /
    • 2006
  • Increasingly complex tasks are performed by computers or cellular phone, requiring more and more memory capacity as well as faster and faster processing speeds. This leads to a constant need to develop more highly integrated circuit systems. Therefore, there have been numerous studies by many engineers investigating circuit patterning. In particular, PCB including module/package substrates such as FCB (Flip Chip Board) has been developed toward being low profile, low power and multi-functionalized due to the demands on miniaturization, increasing functional density of the boards and higher performances of the electric devices. Imprint lithography have received significant attention due to an alternative technology for photolithography on such devices. The imprint technique. is one of promising candidates, especially due to the fact that the expected resolution limits are far beyond the requirements of the PCB industry in the near future. For applying imprint lithography to FCB, it is very important to control thermal properties and mechanical properties of dielectric materials. These properties are very dependent on epoxy resin, curing agent, accelerator, filler and curing degree(%) of dielectric materials. In this work, the epoxy composites filled with silica fillers and cured with various accelerators having various curing degree(%) were prepared. The characterization of the thermal and mechanical properties wasperformed by thermal mechanical analysis (TMA), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), rheometer, an universal test machine (UTM).

  • PDF

Characterization of the molecular and biological properties between the equine herpesvirus type 1 immediate-early protein and the general transcription factor human TFIIB

  • Jang Hyung-Kwan
    • 한국동물위생학회지
    • /
    • 제27권4호
    • /
    • pp.355-369
    • /
    • 2004
  • The equine herpesvirus type 1 (EHV-1) immediate-early (IE) protein is a potent transactivator responsible for the activation of both early and late genes during the course of infection and is comprised of discrete functional domains that mediate its many functions. Interaction between trans activators such as the IE protein and various components of the RNA polymerase II transcription initiation machinery has been demonstrated to be critical for transactivation. In the present report, it is addressed the hypothesis that the IE protein interacts with various components of transcription machinery to mediate transactivation of target viral genes. In these studies, it is demonstrated that in vitro transcribed and translated IE protein interacts with TFIIB-agarose conjugate but not with TFIID-agarose conjugate. Additional immunoprecipitation studies using nuclear extracts derived from EHV-1 infected RK-13 cells confirmed that the IE protein interacts strongly with TFIIB, but fails to interact with TFIID. IR2, a truncated form of the IE protein lacking the potent transactivation domain and involved in the down-regulation of the IE gene, also interacted with TFIIB but not with TFIID. Studies were also performed to ascertain if particular TBP-associated factors (TAFs) could mediate IE or IR2 binding to TFIID. In vitro transcribed and translated TAF250 added to nuclear extracts generated from EHV-1 infected cells also failed to mediate an interaction between the IE protein or the IR2 protein and TFIID. This study demonstrated that the IE protein mediates transactivation of target viral genes by a mechanism that involves TFIIB. This is in contrast to mechanisms that have been proposed for both the herpes simplex virus ICP4 and VP16 protein which have been proposed to transactivate viral genes through interactions involving both TFIIB and TFIID. This study also intimates that IR2 mediate its repressive effects during the course of EHV-1 infection by a mechanism that involves sequestration of various transcription factors.

Microbial Rhodopsins: Genome-mining, Diversity, and Structure/Function

  • Jung, Kwang-Hwan;Vishwa Trivedi;Yang, Chii-Shen;Oleg A. Sineschekov;Elena N. Spudich;John L. Spudich
    • Journal of Photoscience
    • /
    • 제9권3호
    • /
    • pp.45-48
    • /
    • 2002
  • Microbial rhodopsins, photoactive 7-transmembrane helix proteins that use retinal as their chromophore, were observed initially in the Archaea and appeared to be restricted to extreme halophilic environments. Our understanding of the abundance and diversity of this family has been radically transformed by findings over the past three years. Genome sequencing of cultivated microbes as well as environmental genomics have unexpectedly revealed archaeal rhodopsin homologs in the other two domains of life as well, namely Bacteria and Eucarya. Organisms containing these homologs inhabit such diverse environments as salt flats, soil, freshwater, and surface and deep ocean waters, and they comprise a broad phylogenetic range of microbial life, including haloarchaea, proteobacteria, cyanobacteria, fungi, and algae. Analysis of the new microbial rhodopsins and their expression and structural and functional characterization reveal that they fulfill both ion transport and sensory functions in various organisms, and use a variety of signaling mechanisms. We have obtained the first crystallographic structure for a photosensory member of this family, the phototaxis receptor sensory rhodopsin II (SRII, also known as phoborhodopsin) that mediates blue-light avoidance by the haloarchaeon Natronobacterium pharaonis. The structure obtained from x-ray diffraction of 3D crystals prepared in a cubic lipid phase reveals key features responsible for its spectral tuning and its sensory function. The mechanism of SRII signaling fits a unified model for transport and signaling in this widespread family of phototransducers.

  • PDF

인지질막 결합에 필요한 제5혈액응고인자 트립토판잔기들의 역할규명 (Characterization of tryptophan residues of human coagulation factor V required for binding to phospholipid membranes)

  • Kim, Suhng-Wook
    • 생명과학회지
    • /
    • 제13권4호
    • /
    • pp.463-472
    • /
    • 2003
  • 제5인자와 지질막 phosphatidylserine과의 상호작용은 prothrombinase 복합체의 활성을 조절하는데 중요하다. 본 연구에서 제5인자의 지질 결합부위에 위치한 Trp2063과 Trp2064를 동시에 돌연변이 시킨 재조합 제5인자를 과발현 시키고 정제하였다. 돌연변이된 제5인자는 1-10%의 phosphatidylserine을 포함하는 지질막에서 아주낮은 활성을 보였다. surface plasmon resonance에 의해서 지질막과의 결합을 측정한 결과 돌연변이된 제5인자가 본래의 제5인자보다 고정된 지질막에의 결합이 현저하게 떨어지는 것을 관찰하였다. 제5인자가 phosphatidylserine을 포함하는 지질막에 높은 친화력으로 결합하기 위해서는 Trp2063과 Trp2064가 필수적이고 이러한 상호작용은 생리적인 phosphatidylserine 농도를 포함하는 지질막 위에서 prothrombinase 복합체의 형성에 필요하다는 결론을 내렸다.

저온 열처리 목재 표면의 분광학적 특성 (Spectroscopic Characterization of Wood Surface Treated by Low-Temperature Heating)

  • 김강재;나기백;류지애;엄태진
    • Journal of the Korean Wood Science and Technology
    • /
    • 제46권3호
    • /
    • pp.285-296
    • /
    • 2018
  • ISPM No. 15에 의거한 열처리 목재의 검증을 위한 연구로서 열처리 목재 표면의 분광학적 특성을 분석하였다. IR 스펙트럼 상에서는 다양한 작용기들이 확인되지만 수종간, 열처리 시간 및 보관 기간에 따른 특별한 차이를 발견하기 어려웠다. HBI(hydrogen-bonding intensity)는 보관 기간에 따른 열처리 목재의 변화는 관찰할 수 있지만 열처리 시간에 따른 변화는 관찰하기 어려웠다. 하지만 PCA score plot 상에서 수종 내에서 60분 혹은 90분의 열처리 시간에 따라 목재의 분류가 가능하였다. PCA에서 열처리 목재의 분류의 기준은 목재 내 리그닌의 방향족 환과 셀룰로오스의 C-H bending이었으며 이들 성분에 의해 ISPM No. 15로 열처리된 목재를 분류할 수 있었다.

Characterization of a New Anti-dementia β-secretase Inhibitory Peptide from Arctoscopus japonicus

  • Park, Seul Bit Na;Kim, Sung Rae;Byun, Hee-Guk
    • 한국키틴키토산학회지
    • /
    • 제23권4호
    • /
    • pp.220-227
    • /
    • 2018
  • Amyloid plaque is a product of aggregation of ${\beta}$-amyloid peptide ($A{\beta}$) and is an important factor in the pathogenesis of Alzheimer's Disease (AD). $A{\beta}$ is a major component of amyloid plaque and vascular deposits in the AD brain. The enzyme ${\beta}$-secretase is required for the production of $A{\beta}$; thus, prevention of the formation of $A{\beta}$ through the inhibition of ${\beta}$-secretase is a major focus in the study of the treatment of AD. In this study, we investigated ${\beta}$-secretase inhibitory activity of an Arctoscopus japonicus peptide. An Alcalase hydrolysate had the highest ${\beta}$-secretase inhibitory activity. A ${\beta}$-secretase inhibitory activity peptide was separated using ion exchange column chromatography (carboxy-methyl: CM, quaternary methyl ammonium: QMA) and reverse phase high performance liquid chromatography (RP-HPLC) on a C18 column. The $IC_{50}$ value of the purified peptide was $248.2{\pm}1.73{\mu}g/mL$. The ${\beta}$-secretase inhibitory peptide was identified as a six amino acid residue of Gly-Pro-Val-Gly-Ala-Pro (MW: 497.27 Da). In cell viability experiments, the final purified fraction, the carboxy-methyl ion exchange column fraction (CM-F1) showed no significant cytotoxic effect in SH-SY5Y cells at concentrations below $100{\mu}g/mL$ in 24 h. The results of this study suggest that peptides separated from Arctoscopus japonicus may be beneficial as ${\beta}$-secretase inhibitor compounds in functional foods.

방사선 이용 항균성 천연추출물 함유 하이드로겔 제조 및 특성 연구 (Radiation Preparation and Characterization of Antimicrobial Hydrogels)

  • 권희정;박은지;최종배;임종영;정진오;신영민;정성린;박종석;임윤묵
    • 방사선산업학회지
    • /
    • 제7권1호
    • /
    • pp.87-91
    • /
    • 2013
  • Taraxacum platycarpum (TP) has been used for years without restriction on usual dose for its non-toxic nature and nonexistence of the side effects. To develop antimicrobial hydrogel, poly (vinylalcohol) (PVA) hydrogels containing the aqueous extracted TP as an antimicrobial agent were prepared by using gamma-rays irradiation. The antimicrobial activities of the TP hydrogels were tested against Staphylococcus aureus and Staphylococcus epidermidis by disc diffusion method. The inhibition zones (IZ) of the TP extracts and TP hydrogels against S. aureus were 16 mm and 20 mm and against S. epidermidis was 10 mm and 13 mm, respectively. In conclusion, the TP hydrogel that has an excellent antimicrobial activity was proved to be a valuable material for functional skin care.