Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Production, Purification, and Characterization of Phaseolus vulgaris Leghemoglobin a in Pichia pastoris

Pichia pastoris에서 강낭콩 Leghemoglobin a의 생산, 정제 및 특징

  • Jun-Young, Kim (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering,Dong-Eui University) ;
  • Da-Hee, Han (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering,Dong-Eui University) ;
  • Geun-o, Park (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering,Dong-Eui University) ;
  • Soo-Wan, Nam (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering,Dong-Eui University) ;
  • Yeon-Hee, Kim (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering,Dong-Eui University) ;
  • Han-Woo, Kim (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Sung-Jong, Jeon (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering,Dong-Eui University)
  • 김준영 (동의대학교 바이오응용공학부 의생명공학전공) ;
  • 한다희 (동의대학교 바이오응용공학부 의생명공학전공) ;
  • 박근오 (동의대학교 바이오응용공학부 의생명공학전공) ;
  • 남수완 (동의대학교 바이오응용공학부 의생명공학전공) ;
  • 김연희 (동의대학교 바이오응용공학부 의생명공학전공) ;
  • 김한우 (극지연구소 생명과학연구본부) ;
  • 전숭종 (동의대학교 바이오응용공학부 의생명공학전공)
  • Received : 2022.11.02
  • Accepted : 2022.11.21
  • Published : 2022.12.28
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Abstract

In this study, Phaseolus vulgaris (kidney bean) leghemoglobin a (PhLba) gene was cloned into pPICZαA and expressed in Pichia pastoris to sustainably produce a heme-carrying protein for organoleptic use in plant-based meat. The recombinant PhLba protein was secreted into the culture medium in a solubilized form, and the molecular weight of the purified PhLba was estimated to be 16.5 kDa using SDS-PAGE. In addtion, the yield of recombinant PhLba holoprotein was enhanced by supplementation of the cultivation medium with hemin. This result indicates that the apo-forms of PhLba can be effectively saturated with cofactor.

식물성 육류에서 미각적인 자극을 얻기 위해 사용하는 heme 함유 단백질의 지속적인 생산을 위해 강낭콩 유래 leghemoglobin a (PhLba) 유전자를 pPICZαA에 클로닝하고 Pichia pastoris에서 발현하였다. 재조합 PhLba 단백질은 가용화 형태로 배양 배지속으로 분비되었다. 정제된 PhLba의 분자량은 SDS-PAGE 상에서 16.5 kDa으로 나타났다. 재조합 PhLba holoprotein의 수율은 배양 배지에 hemin을 첨가함으로써 향상되었다. 이것은 PhLba의 apo 형태가 보조인자와 함께 효과적으로 포화된다는 것을 나타낸다.

Keywords

Acknowledgement

This work was supported by Dong-Eui University Grant(202201710001).

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