• 제목/요약/키워드: YebF

검색결과 12건 처리시간 0.021초

Co-Expression of a Chimeric Protease Inhibitor Secreted by a Tumor-Targeted Salmonella Protects Therapeutic Proteins from Proteolytic Degradation

  • Quintero, David;Carrafa, Jamie;Vincent, Lena;Kim, Hee Jong;Wohlschlegel, James;Bermudes, David
    • Journal of Microbiology and Biotechnology
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    • 제28권12호
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    • pp.2079-2094
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    • 2018
  • Sunflower trypsin inhibitor (SFTI) is a 14-amino-acid bicyclic peptide that contains a single internal disulfide bond. We initially constructed chimeras of SFTI with N-terminal secretion signals from the Escherichia coli OmpA and Pseudomonas aeruginosa ToxA, but only detected small amounts of protease inhibition resulting from these constructs. A substantially higher degree of protease inhibition was detected from a C-terminal SFTI fusion with E. coli YebF, which radiated more than a centimeter from an individual colony of E. coli using a culture-based inhibitor assay. Inhibitory activity was further improved in YebF-SFTI fusions by the addition of a trypsin cleavage signal immediately upstream of SFTI, and resulted in production of a 14-amino-acid, disulfide-bonded SFTI free in the culture supernatant. To assess the potential of the secreted SFTI to protect the ability of a cytotoxic protein to kill tumor cells, we utilized a tumor-selective form of the Pseudomonas ToxA (OTG-PE38K) alone and expressed as a polycistronic construct with YebF-SFTI in the tumor-targeted Salmonella VNP20009. When we assessed the ability of toxin-containing culture supernatants to kill MDA-MB-468 breast cancer cells, the untreated OTG-PE38K was able to eliminate all detectable tumor cells, while pretreatment with trypsin resulted in the complete loss of anticancer cytotoxicity. However, when OTG-PE38K was co-expressed with YebF-SFTI, cytotoxicity was completely retained in the presence of trypsin. These data demonstrate SFTI chimeras are secreted in a functional form and that co-expression of protease inhibitors with therapeutic proteins by tumor-targeted bacteria has the potential to enhance the activity of therapeutic proteins by suppressing their degradation within a proteolytic environment.

CT형 점용접 이음재의 피로설계에 관한 연구 (A Study on Fatigue Design of CT-Type Spot Welded Lap Joint)

  • 백승엽
    • Journal of Welding and Joining
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    • 제28권2호
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    • pp.91-95
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    • 2010
  • Stress distribution and deformation on the CT-type(Cross Tension type) spot welded lap joint subjected to out of plane tensile load were investigated by finite element method. Using the maximum principal stresses at the nugget edge obtained by FEM analysis, evaluated the fatigue strength of the CT-type spot welded lap joints having various dimensions and materials. and also, the influence of the geometrical parameters of CT-type spot welded lap joints on stress distribution and fatigue strength must be evaluated. thus, in this paper, ${\Delta}P-N_f$ curve were obtained by fatigue tests. Using these results, ${\Delta}P-N_f$ curve were systematically rearranged in the $\Delta\sigma-N_f$ relation with the hot spot stresses at the CT-type spot welded lab joints. It was found that the proposed $\Delta\sigma-N_f$ relation could provide a more reasonable fatigue design criterion for the CT-type spot welded lap joints.

STS301L 용접종류별 이음재의 피로설계에 관한 연구 (A Study on Fatigue Design for Welded Joint of STS301L)

  • 백승엽
    • Journal of Welding and Joining
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    • 제28권3호
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    • pp.86-91
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    • 2010
  • Stainless steel sheets are widely used as the structural material for the railroad cars and the commercial vehicles. These kinds structures used stainless steel sheets are commonly fabricated by using the gas welding. For fatigue design of gas welded joints such as fillet and plug type joint, it is necessary to obtain design information on stress distribution at the weldment as well as fatigue strength of gas welded joints. And also, the influence of the geometrical parameters of gas welded joints on stress distribution and fatigue strength must be evaluated. the ${\Delta}P-N_f$ curves were obtained by fatigue tests. Using these results, ${\Delta}P-N_f$ curves were rearranged in the ${\Delta}{\sigma}-N_f$ relation with the maximum stress at the edge of fillet welded joint.

STS301L 용접이음재의 피로설계에 관한 연구 (A Study on Fatigue Design for Welded Joint of STS301L)

  • 백승엽
    • 한국자동차공학회논문집
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    • 제18권4호
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    • pp.127-131
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    • 2010
  • Stainless steel sheets are widely used as the structural material for the railroad cars and the commercial vehicles. These kinds of structures used stainless steel sheets are commonly fabricated by using the gas welding. For fatigue design of gas welded joints such as fillet and plug type joint, it is necessary to obtain design information on stress distribution at the weldment as well as fatigue strength of gas welded joints. And also, the influence of the geometrical parameters of gas welded joints on stress distribution and fatigue strength must be evaluated. Thus, in this paper, ${\Delta}P-N_f$ curves were obtained by fatigue tests. Using these results, ${\Delta}P-N_f$ curves were rearranged in the. ${\Delta}{\sigma}-N_f$ relation with the hot spot stresses at the gas welded joints.

STS 301L 필렛 용접이음재의 피로설계에 관한 연구 (A Study on Fatigue Design of STS301L Fillet Welded Joint)

  • 백승엽
    • 대한기계학회논문집A
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    • 제34권5호
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    • pp.561-565
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    • 2010
  • 용접부는 일반적으로 잘 알려져 있는 바와 같이, 외력에 의한 응력 집중원(stress concentration source)이 되는 것은 물론 용접과정에서의 입열(heat input)로 인한 성분과 조직의 변화에 의해 반복하중에 대한 피로강도(fatigue strength)가 모재의 그것에 비해 훨씬 떨어져서 피로균열(fatigue crack)의 발단이 되고 있다. 따라서, 본 연구에서는 실제로 철도차량에 적용되는 가스용접 이음재 가운데 대표적인 형상시편들을 재질별로 피로실험을 수행하여 피로하중범위-피로수명(${\Delta}P-N_f$) 관계를 도출하여 비교 평가하였다. 또한, 수치 해석적 유한요소법을 이용하여 용접이음재의 응력분포를 해석한 후, 용접이음부에 발생하는 최대주응력으로 (${\Delta}P-N_f$) 관계를 $\Delta\sigma-N_f$ 관계로 재정리하였다. 이상의 결과들을 바탕으로, 가스용접이 적용된 철도차량 차체의 경제적이고 합리적인 피로설계를 위한 기초정보로 사용하고자 하였다.

유한요소법을 이용한 파괴 역학적 방법의 신뢰성설계기술에 관한 연구 (A Study on Reliability Design of Fracture Mechanics Method Using FEM)

  • 백승엽;이봉구
    • 한국산학기술학회논문지
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    • 제16권7호
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    • pp.4398-4404
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    • 2015
  • 스테인리스 강판은 동적기계구조물을 제작하기 위한 구조용 재료로 널리 사용된다. 또한 이들을 일체화 시키는 방법으로 가스용접을 많이 사용하게 되는데 가스용접에는 다양한 종류에 의해 구조물을 일체화 시킨다. 따라서 부재와 부재를 연결하는 용접부에 대한 응력분포 및 피로강도평가는 구조물의 건전성 및 수명을 연구하는데 매우 중요한 요소가 된다. 그래서 본 연구에서는 피로시험에 의해서 얻어지는 ${\Delta}P-N_f$ 관계를 유한요소해석법에 의해서 최대주응력으로 ${\Delta}{\sigma}-N_f$ 관계로 나타내어 피로설계기준을 정하였고, 이 결과를 이용해서 확률론적 통계해석기법을 적용해서 가속식을 추정하여 임의의 목표수명을 예측할 수 있는 신뢰성설계기술기법을 제시하고자 하였다.

STS301L 가스용접 이음재의 가속수명예측 (I) - Fillet Type - (Accelerated Life Prediction for STS301L Gas Welded Joint (I) - Fillet Type -)

  • 백승엽
    • 대한기계학회논문집A
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    • 제34권4호
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    • pp.467-474
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    • 2010
  • 용접이음부의 신뢰성 확보는 구조물의 건전성과 내구성에 직접적인 영향을 미치기 때문에 가스용접 이음재의 피로설계기준(fatigue design criterion)을 정하기 위해서는 피로시험을 수행하여 ${\Delta}P$-Nf 관계를 이용하는 것이 일반적이다. 그러나 피로데이터를 장시간 획득하는 과정에서 여러 가지 변동인자에 의해서 피로데이터가 영향을 받기 때문에 피로데이터의 신뢰도가 떨어진다. 또한, 이음재의 재질 및 접합형태가 달라질 때마다, 각각의 경우에 대해서 새로운 피로시험이 요구됨으로 많은 시간과 비용이 소모된다. 따라서 이러한 문제점들을 개선하고 신뢰성 있는 설계를 하기 위해서 필렛 가스용접 이음재를 적용, 반복피로시험을 통한 데이터를 통계적으로 분석하여, 다양한 가스용접 이음재의 피로수명을 예측함과 동시에 실제 피로시험데이터와 비교 분석하여 예측된 수명의 신뢰도(reliability)와 신뢰구간(confidence interval)을 추정함으로서 새로운 피로설계기준 방법을 제시하고자 하였다.

STS301L 가스용접이음재의 가속수명에측에 관한 연구 (1. Plug and Ring type) (A study on Accelerated Life Prediction of Gas Welded joint of STS301L (1. Plug and Ring type))

  • 백승엽;배동호
    • 대한기계학회:학술대회논문집
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    • 대한기계학회 2008년도 추계학술대회A
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    • pp.1355-1360
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    • 2008
  • Stainless steel sheets are widely used as the structure material for the railroad cars and the commercial vehicles. These kinds structures used stainless steel sheets are commonly fabricated by using the gas welding. Gas welding is very important and useful technology in fabrication of an railroad car and vehicles structure. However fatigue strength of the gas welded joints is considerably lower than parent metal due to stress concentration at the weldment, fatigue strength evaluation of gas welded joints are very important to evaluate the reliability and durability of railroad cars and to establish a criterion of long life fatigue design. In this paper, ${\Delta}P-N_f$ curve were obtained by fatigue tests. Using these results, the accelerated life test (ALT) is conducted. From the experimental results, an acceleration model is derived and acceleration factors are estimated. So it is intended to obtain the useful information for the fatigue lifetime of plug and ring gas welded joints and data analysis by statistic reliability method, to save time and cost, and to develop optimum accelerated life prediction plans.

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철도차량 차체 가스용접 이음재의 가속수명예측과 신뢰도 평가 (Reliability Assessment and Accelerated Life Prediction of Gas Welded Joint in the Rail Road Car Body (1. Plug and Ring Type))

  • 백승엽
    • Journal of Welding and Joining
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    • 제28권1호
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    • pp.77-85
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    • 2010
  • Stainless steel sheets are widely used as the structure material for the railroad cars and the commercial vehicles. These kinds structures used stainless steel sheets are commonly fabricated by using the gas welding. Gas welding is very important and useful technology in fabrication of a railroad car and vehicles structure.However fatigue strength of the gas welded joints is considerably lower than parent metal due to stress concentration at the weld, fatigue strength evaluation of gas welded joints are very important to evaluate the reliability and durability of railroad cars and to establish a criterion of long life fatigue design. In this paper, $({\Delta}{\sigma}_a)_R-N_f$ curve were obtained by fatigue tests. Using these results, the accelerated life test(ALT) was conducted. From the experimental results, an acceleration model was derived and acceleration factors are estimated. So it is intended to obtain the useful information for the fatigue lifetime of plug and ring gas welded joints and data analysis by statistic reliability method, to save time and cost, and to develop optimum accelerated life prediction plans.

STS301L 가스용접이음재의 가속수명예측 자동화에 관한 연구 (Plug and Ring Type) (A Study on Accelerated Life Prediction Automation of Gas Welded Joint of STS301L (Plug and Ring Type))

  • 백승엽;손일선
    • 한국자동차공학회논문집
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    • 제19권3호
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    • pp.1-8
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    • 2011
  • Stainless steel sheets are widely used as the structure material for the railroad cars and the commercial vehicles. These kinds structures used stainless steel sheets are commonly fabricated by using the gas welding. Gas welding is very important and useful technology in fabrication of an railroad car and vehicles structure. However fatigue strength of the gas welded joints is considerably lower than parent metal due to stress concentration at the weldment, fatigue strength evaluation of gas welded joints are very important to evaluate the reliability and durability of railroad cars and to establish a criterion of long life fatigue design. In this paper, ${\Delta}-N_f$ curve were obtained by fatigue tests. Using these results, the accelerated life test (ALT) is conducted. From the experimental results, an acceleration model is derived and acceleration factors are estimated. So it is intended to obtain the useful information for the fatigue lifetime of plug and ring gas welded joints and data analysis by statistical reliability method, to save time and cost, and to develop optimum accelerated life prediction plans.