• 제목/요약/키워드: Metabolic Engineering

검색결과 747건 처리시간 0.035초

Characterization of L-(+)-Lactic Acid Producing Weizmannia coagulans Strains from Tree Barks and Probiogenomic Evaluation of BKMTCR2-2

  • Jenjuiree Mahittikon;Sitanan Thitiprasert;Sitanan Thitiprasert;Naoto Tanaka;Yuh Shiwa;Nitcha Chamroensaksri;Somboon Tanasupawat
    • 한국미생물·생명공학회지
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    • 제51권4호
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    • pp.403-415
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    • 2023
  • This study aimed to isolate and identify L-(+)-lactic acid-producing bacteria from tree barks collected in Thailand and evaluate the potential strain as probiotics. Twelve strains were isolated and characterized phenotypically and genotypically. The strains exhibited a rod-shaped morphology, high-temperature tolerance, and the ability to ferment different sugars into lactic acid. Based on 16S rRNA gene analysis, all strains were identified as belonging to Weizmannia coagulans. Among the isolated strains, BKMTCR2-2 demonstrated exceptional lactic acid production, with 96.41% optical purity, 2.33 g/l of lactic acid production, 1.44 g/g of lactic acid yield (per gram of glucose consumption), and 0.0049 g/l/h of lactic acid productivity. This strain also displayed a wide range of pH tolerance, suggesting suitability for the human gastrointestinal tract and potential probiotic applications. The whole-genome sequence of BKMTCR2-2 was assembled using a hybridization approach that combined long and short reads. The genomic analysis confirmed its identification as W. coagulans and safety assessments revealed its non-pathogenic attribute compared to type strains and commercial probiotic strains. Furthermore, this strain exhibited resilience to acidic and bile conditions, along with the presence of potential probiotic-related genes and metabolic capabilities. These findings suggest that BKMTCR2-2 holds promise as a safe and effective probiotic strain with significant lactic acid production capabilities.

산단지역 공기 중 휘발성유기화합물농도와 지역주민의 노출 수준 (Concentration of volatile organic compounds(VOCs) in ambient air and level of residents in industrial area)

  • 우경숙;박희진;강택신;김근배;전준민;장봉기;이종화;손부순
    • 한국산업보건학회지
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    • 제25권1호
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    • pp.104-114
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    • 2015
  • Objectives: The purpose of this study is to investigate the atmospheric concentration of VOCs and the urinary concentration of t,t-MA, HA, MA in the industrial complex of Yeosu, South Jeolla Province. Methods: In order to study seasonal patterns of air concentration of VOCs, measurements were taken at five sampling sites around Yeosu from June 2013 to June 2014. Urinary metabolite excretionsfrom 671 subjects, exposure and comparison area were analyzed. Results: The average concentration of VOCs in the air was 1.53ppb for benzene, 0.73ppb for toluene, 0.22ppb for ethylbenzene, 0.52ppb for xylene and 0.12ppb for styrene. The concentration of benzene was somewhat higher than the year-average standard ($5{\mu}g/m^3$, about 1.5ppb) of the domestic air-environment criteria newly established in 2010.The metabolic concentration of VOCs in the urine of the entire sample was analyzed at $47.76{\mu}g/g\;cr.$, 213.07mg/g cr., and $290.09{\mu}g/g\;cr.$ for t,t-MA, HA, and MA, respectively. Compared with the average values for Korea as presented in the first basic survey of national environmental conservation ( $49.8{\mu}g/g\;cr.$ for t,t-MA, 0.17g/g cr. for HA, and 0.26mg/g cr. for MA), the metabolic concentrations of HA and MA in urine were higher than the average values. Conclusions: The concentration of VOCs in the air and urinary metabolites of the exposed and control areas showed that the concentrations of all substances were higher in the exposed area than in the control area.

Genenation of structural diversity in polyketides by combinatorial biosynthesis of polyketides: Part I. Generation of multiple bioactive macrolides by hybrid modular polyketide synthases in Streptomyces venezuelae, Part II. Production of novel rifamycins by combinatorial biosynthesis

  • Yoon, Yeo-Joon
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2002년도 학술발표대회
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    • pp.18-25
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    • 2002
  • The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

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간호교육에서의 유전학 교육과정 현황과 요구 (The Current Status and Educational Requirements for Genetics Curriculum at Nursing Institutions)

  • 홍해숙;변영순;나연경
    • Journal of Korean Biological Nursing Science
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    • 제5권1호
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    • pp.13-22
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    • 2003
  • The purpose of this study was to investigate and analyze current educational requirements related to genetics curriculum(from June 2002 to September 2002) established at nursing institutions and to provide the basic data for the development of genetics science program at the undergraduate. Subjects of this study were comprised of twenty-three colleges of nursing in 4-year baccalaureate and thirty colleges in 3-year diploma programs. The results of this study were as follows : 1) 32 colleges offer courses related to genetics. 29 among 32 colleges have that integrated. Three schools have established completely independent courses of genetics. 21 colleges do not have any courses dealing with genetics. 2) The contents of courses related to genetics include: Congenital abnormalities, chromosomal aberrations, congenital metabolic disease, prenatal diagnosis and genetic counseling, genes and chromosomes, immune genetics, blood type and genetics, rule of genetics, variation in gene expression, the map of the human gene, gene linkage genetics, interaction of genes, single inheritance in order and genetic biochemistry. 3) For course credit, 14colleges(48.3%) offered at most 1 credit per course. The grade of student who can take the course, 51.7% were in their second year while 37.9% were in their third year. The majors of nursing faculty who taught the course were nursing(51.7%) and basic nursing science(17.2%). 4) As far as the need of opening the courses related to genetics, 36 colleges(67.0%) have made a 'need', 12 schools(22.6%) state 'dose not need'. 711e reason for need were the following development of bio engineering, increase number of patients who are related to genetics, recognition of the need in clinical nursing. 7 schools(13.2%) agreed to offer independent course in genetics but 39 schools(73.6%) are in disagreement with that. When the school offers the course with other courses, 27 schools(50.0%) are opening basic nursing science and 14 schools(26.4%) are opening nursing as an integrated courses. If the name of course was either genetic nursing(34.0%) or genetics(28.3%), the credits for the course was one or 2 credits. 33 schools(62.3%) students were in the first or second years. 41 schools(84.9%), the majors of the faculty who had taught the course were either basic nursing science(35.8%), nursing(28.3%) or basic medicine(24.5%). The contents of the course should include in that order: Chromosome aberrations, prenatal diagnosis and genetic counseling, congenital metabolic disease, congenital abnormalities, genes and chromosomes, the rules of genetics, immune genetics, interaction of genes, variation in gene expression, etc. The results and discussions of the study indicate that the entire curriculums need to be investigated with respect to contents of education, nursing curriculums and name of courses because of the increasing need of knowledge related to genetics in the clinical practice.

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인($^{31}$P) 자기공명분광법을 사용하여 사립체 근질병환자와 정상인과의 대사물질 비교조사 (Metabolic Abnormalities in Patients with Mitochondrial Myopathy Evaluated by In Vivo $^{31}$P Magnetic Resonance Spectroscopy)

  • Bo-Young Choe
    • Investigative Magnetic Resonance Imaging
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    • 제2권1호
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    • pp.89-95
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    • 1998
  • 목적 : 인($^{31}P$) 자기공명분광법을 사용하여 사립체 근병(mitochondria myopathy) 환자의 대퇴부 근조직의 대사물질의 변화를 정상인과 비교조사하였다. 대상 및 방법 : 사립체 근병환자 10명과 정상인 10명을 대상으로 1.5T MRI/MRS 장비를 사용하여 인($^{31}P$) 자기공명분광법을 적용하였다. 오른쪽 대퇴부위의 근조직에 $4{\;}{\times}{\;}4{\;}{\times}4{\;}cm^{3}$ 의 관심부위 (volume of interest ; VOI)를 선정하여 image selected in vivo spectroscopy (ISIS)를 저용하였다. 인대사불질의 정\ulcorner분석은 Marquart algorithm을 사용하였다. 결과 : 사립체 근병환자의 특징은 정상인과 비교하여 Pe/PCr 대사비율이 상당히 증가하고 (P=0.003), ATP/PCr 대사비율은 상당히 감소하였다(p=0.004). 특히 ATP 중 ${\beta}-ATP/PCr$ 비율의 변화가 가장 심하게 나타났다. 환자군과 정상군의 pH 차이는 통계학적으로 큰 의의는 없었다. 결론 : 인($^{31}P$) 자기 공명분광법은 사립체 근병환자의 대퇴부 근조직의 ATP/PCr 과 Pi/PCr 대사비율을 토대로 유용한 임상 평가 자료를 제공하고, 따라서 근대사물질의 질병을 이해하는데 도움을 줄 것으로 사료된다.

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Next-generation probiotics, parabiotics 및 postbiotics (Next-generation Probiotics, Parabiotics, and Postbiotics)

  • 조광근;이승호;최인순;이상원
    • 생명과학회지
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    • 제31권6호
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    • pp.595-602
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    • 2021
  • 사람의 장내 미생물은 숙주의 대사 조절에서 중요한 역할을 한다. 장내 미생물 균총의 불균형은 비만, 대사 증후군과 밀접하게 병리학적, 생리학적 상호작용을 한다. A. muciniphila는 최근 인간의 대변에서 분리되었으며, 장내 미생물의 약 1-4%를 차지하는 우점균이다. A. muciniphila 유래 외막(external membran) 단백질 Amuc_1100과 세포외 소포(EVs)의 이용은 비만치료를 위한 새로운 전략이 될 수 있다. A. muciniphila는 비만과 같은 대사 장애치료를 위한 차세대 probiotics로 고려되고 있다. F. prausnitzii는 next-generation probiotic로서 건강한 성인의 경우 장내미생물 균총의 약 5%를 차지하며 성인 장 건강의 지표이다. F. prausnitzii는 butyrate-producing bacterium로서 항염증 효과를 나타내며, 면역 질환과 당뇨병 치료를 위한 next-generation probiotics로 기대된다. Postbiotics는 probiotics에 의해 분비되는 세포 상층액에 포함된 대사산물의 복잡한 혼합물이다. 반면, parabiotics는 probiotics를 불활성화 시킨 미생물 세포이다. Paraprobiotics와 postbiotics는 probiotics에 비해 명확한 화학구조(clear chemical structures)와 안전한 투여 용량(safety dose parameters), 장기간의 유통 기간(longer shelf life) 등 많은 장점을 가지고 있어서 probiotics를 대체할 수 있는 잠재력을 가지고 있다. 장 생태계의 불균형을 정상적으로 복원하기 위해서 장내 공생 박테리아(commensal bacteria) 중 next-generation probiotics (NGPs)를 사용하는 것이 가장 자연스런 방법이다. 따라서 next-generation probiotics 를 대상으로 parabiotics와 postbiotics와 같은 새로운 식품이나 약품으로 개발하는 것이 필요하다.

PET Imaging of Click-engineered PSMA-targeting Immune Cells in Normal Mice

  • Hye Won Kim;Won Chang Lee;In Ho Song;Hyun Soo Park;Sang Eun Kim
    • 대한방사성의약품학회지
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    • 제8권2호
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    • pp.53-61
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    • 2022
  • This study aimed to increase the targeting ability against PSMA in cell therapy using metabolic glycoengineering and biorthogonal chemistry and to visualize cell trafficking using PET imaging. Cellular membranes of THP-1 cells were decorated with azide(-N3) using Ac4ManNAz by metabolic glycoengineering. Engineered THP-1 cells were conjugated with DBCO-bearing fluorophore (ADIBO-Cy5.5) for 1 h at different concentrations and analyzed by confocal fluorescence microscopy and flow cytometry. For PSAM ligand conjugation to THP-1 cells, Ac4ManNAz treated THP-1 cells were incubated with DBCO-PSMA ligand (ADIBO-GUL) at a final concentration with 100 µM for 1 h. To evaluate the effect on cell recognition, PSMA ligand conjugated THP-1 cells(as effectors) were co-cultured with PSMA positive 22RV1 (as target cells) at 3 : 1 a effector-to-target cell (E/T) ratio. The interaction between THP-1 and 22RV1 was monitored by confocal fluorescence microscopy. For preparing the radiolabeled THP-1, the cells were treated at the activity of ~ 740 kBq of [89Zr]Zr(oxinate)4/5 × 106 cells. Radiolabeled cells were analyzed for determination of cell-associated radioactivity by gamma counting and viability using MTS assay. In the cytotoxicity assay, THP-1 cells did not have any cytotoxicity even when the Ac4ManNAz concentration was 100 µM. In confocal microscopy and flow cytometry, THP-1 cells were efficiently labeled ADIBO-Cy5.5 in a dose-dependent manner, and the dose of 100 µM was the optimal concentration for the following experiments. The clusters of PSMA ligand-conjugated THP-1 cells and 22RV1 cells were identified, indicating cell-cell recognition over the cell surface between two types of cells. Cell radiolabeling efficiency was 54.5 ± 17.8%. THP-1 labeled with 0.09 ± 0.03 Bq/cell showed no significant cytotoxicity compared to unlabeled THP-1 up to 7 days. We successfully demonstrated that Ac4ManNAz treated cells were efficiently conjugated with ADIBO-GUL for preparing the PSMA-targeting cells, and [89Zr]Zr(oxinate)4 could be used to label cells without toxicity. It suggested that PSMA-ligand conjugated cell therapy could be improved cell targeting and be monitored by PET imaging.

Transcription Analysis of Recombinant Trichoderma reesei HJ-48 to Compare the Molecular Basis for Fermentation of Glucose and Xylose

  • Huang, Jun;Lin, Mei;Liang, Shijie;Qin, Qiurong;Liao, Siming;Lu, Bo;Wang, Qingyan
    • Journal of Microbiology and Biotechnology
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    • 제30권10호
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    • pp.1467-1479
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    • 2020
  • Profiling the transcriptome changes involved in xylose metabolism by the fungus Trichoderma reesei allows for the identification of potential targets for ethanol production processing. In the present study, the transcriptome of T. reesei HJ-48 grown on xylose versus glucose was analyzed using next-generation sequencing technology. During xylose fermentation, numerous genes related to central metabolic pathways, including xylose reductase (XR) and xylitol dehydrogenase (XDH), were expressed at higher levels in T. reesei HJ-48. Notably, growth on xylose did not fully repress the genes encoding enzymes of the tricarboxylic acid and respiratory pathways. In addition, increased expression of several sugar transporters was observed during xylose fermentation. This study provides a valuable dataset for further investigation of xylose fermentation and provides a deeper insight into the various genes involved in this process.

Thrichoderma atroviride 배양액의 tyrosinase 억제제에 관한 연구 (Inhibition of Tyrosinase by Metabolites Originating from Thrichoderma atroviride)

  • 강동우;김규민;김예성;서유진;송다영;오다윤;최시온;황주현;김삼웅;방규호;갈상완
    • 생명과학회지
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    • 제31권1호
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    • pp.47-51
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    • 2021
  • 현대인에게 미백화장품은 미를 나타내는 중요한 요소이다. 균류는 미백에 연관된 다양한 물질대사산물을 생산한다. 본 연구에서 미백기능성 화장품에 소재를 탐색하기 위해 Trichoderma atroviride 배양액에 물질대사산물에 의한 tyrosinase inhibitors를 조사하였다. 또한, 억제 효과는 식의약안전처에 미백화장품 원료로 승인된 알부틴과 비교하였다. T. atroviride 배양액에 물질대사산물은 알부틴의 활성보다 높은 tyrosinase 억제 활성을 보였다. T. atroviride 배양액에 포함된 tyrosinase inhibitors 중 일부는 열에 안정한 반면에, 일부는 열에 불안정하였다. 본 물질들은 약산성~중성 pH 영역에서는 안정하였지만, 그 외 영역에서는 매우 불안정하였다. Proteinase K에 의해 활성 영향이 거의 없었기 때문에 inhibitor들은 대부분 물질대사산물로 구성된 것으로 추정된다. 따라서 본 T. atroviride 배양액에 물질대사산물은 미백화장품 소재로써 잠재적 가치가 있는 것으로 제의된다.

키조개(Atrina pectinata)를 이용하여 양식한 흑진주의 특성평가 (The characteristics of black pearl cultured using Pen shell (Atrina pectinata))

  • 이정임;김판채
    • 한국결정성장학회지
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    • 제18권5호
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    • pp.205-210
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    • 2008
  • 진주는 유기질 보석의 일종으로서, 광산에서 발굴하는 보석이 아니라, 연체동물(Mollusc)의 내부에서 생광물화(biomineralization)의 결과로 얻어지는 고부가가치 상품이다. 이러한 진주를 생산해 낼 수 있기 위해서는 연체동물 내부의 패각 안쪽에 진주층(nacre)이 존재해야 하는데, 이에 본 연구에서는 기존의 진주양식에 사용되지는 않지만, 패각 안쪽 진주층이 두텁고 아름다운 색을 지니고 있어 충분히 진주 양식의 가능성을 보이는 키조개(Atrina pectinata)를 이용하여 새로운 양식 진주를 개발하고자 조사, 분석하였다. 주사전자현미경을 통해 키조개(Atrina pectinata)를 모패로 사용하여 양식된 흑진주의 진주층을 관찰한 결과, 이매패 진주층에서 나타나는 Aragonite형 탄산칼슘 층의 특징적인 다각형 판상구조가 관찰되였다. 또한 구체적인 특징을 알아보기 위해 분석한 결과, ED-XRF를 통해 해수 진주의 특징적 원소가 검출되었고, Raman 분석에서는 Aragonite의 특징적 $1083cm^{-1}$, $705cm^{-1}$ peak가 검출되었다. 그러나, UV-Vis 스펙트럼 분석에서는 흑접패(P. margaritifera) 흑진주와는 다른 요인에 기인하는 발색 양상임을 예상하게 했으며 이는 흑진주를 생산하는 모패의 종이 다르기 때문으로 생각된다.