medric medric
[닫기]
잠시만 기다려 주세요. 로딩중입니다.

Expression and Purification of Extracellular Solute-Binding Protein (ESBP) in Escherichia coli, the Extracellular Protein Derived from Bifidobacterium longum KACC 91563

한국축산식품학회지 2019년 39권 4호 p.601 ~ 609
 ( Song Min-Yu ) - National Institute of Animal Science Animal Products Development and Processing Division

 ( Kim Hyae-Kang ) - Seoul National University Department of Agriculture and Biotechnology
 ( Kwak Woo-Ri ) - C&K Genomics
 ( Park Won-Seo ) - National Institute of Animal Science Animal Products Development and Processing Division
 ( Yoo Ja-Yeon ) - National Institute of Animal Science Animal Products Development and Processing Division
 ( Kang Han-Byul ) - National Institute of Animal Science Animal Products Development and Processing Division
 ( Kim Jin-Hyoung ) - National Institute of Animal Science Animal Products Development and Processing Division
 ( Kang Sun-Moon ) - National Institute of Animal Science Animal Products Development and Processing Division
 ( Ba Hoa-Van ) - National Institute of Animal Science Animal Products Development and Processing Division
 ( Kim Bu-Min ) - National Institute of Animal Science Animal Products Development and Processing Division
 ( Oh Mi-Hwa ) - National Institute of Animal Science Animal Products Development and Processing Division
 ( Kim Hee-Bal ) - Seoul National University Department of Agricultural Biotechnology
 ( Ham Jun-Sang ) - National Institute of Animal Science Animal Products Development and Processing Division

Abstract


Bifidobacterium longum KACC 91563 secretes family 5 extracellular solute-binding protein via extracellular vesicle. In our previous work, it was demonstrated that the protein effectively alleviated food allergy symptoms via mast cell specific apoptosis, and it has revealed a therapeutic potential of this protein in allergy treatment. In the present study, we cloned the gene encoding extracellular solute-binding protein of the strain into the histidine-tagged pET-28a(+) vector and transformed the resulting plasmid into the Escherichia coli strain BL21 (DE3). The histidine-tagged extracellular solute-binding protein expressed in the transformed cells was purified using Ni-NTA affinity column. To enhance the efficiency of the protein purification, three parameters were optimized; the host bacterial strain, the culturing and induction temperature, and the purification protocol. After the process, two liters of transformed culture produced 7.15 mg of the recombinant proteins. This is the first study describing the production of extracellular solute-binding protein of probiotic bacteria. Establishment of large-scale production strategy for the protein will further contribute to the development of functional foods and potential alternative treatments for allergies.

키워드

Bifidobacterium longum; probiotics; extracellular vesicle; extracellular solute-binding protein (ESBP); gene cloning
원문 및 링크아웃 정보
등재저널 정보
KCI