Cell surface display of functional macromolecule fusions on Escherichia coli for development of an autofluorescent whole-cell biocatalyst

被引:28
|
作者
Yang, Chao [1 ,2 ]
Zhao, Qiao [4 ,5 ]
Liu, Zheng [1 ,2 ]
Li, Qiyun [3 ]
Qiao, Chuanling [1 ]
Mulchandani, Ashok [6 ]
Chen, Wilfred [6 ]
机构
[1] Chinese Acad Sci, State Key Lab Integrated Management Pest Insects, Inst Zool, Beijing 100101, Peoples R China
[2] Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
[3] Jilin Acad Agr Sci, Inst Plant Protect, Gongzhuling 136100, Peoples R China
[4] Ohio State Univ, Ctr Plant Biotechnol, Columbus, OH 43210 USA
[5] Ohio State Univ, Dept Plant Cellular & Mol Biol, Columbus, OH 43210 USA
[6] Univ Calif Riverside, Dept Chem & Environm Engn, Riverside, CA 92521 USA
关键词
D O I
10.1021/es800441t
中图分类号
X [环境科学、安全科学];
学科分类号
08 ; 0830 ;
摘要
At present, Lpp-OmpA-mediated surface display has opened a new dimension in the development of whole-cell factories. Here, we report the surface display of methyl parathion hydrolase (MPH) and enhanced green fluorescent protein (EGFP) fusions (60 kDa) by employing the Lpp-OmpA chimera as an anchoring motif. A broad-host-range vector, pLOMG33, coding for Lpp-OmpA-MPH-GFP fusion protein was constructed for targeting the fusion protein onto the surface of Escherichia coli. The surface localization of fusion protein was demonstrated by Western blot analysis, immunofluorescence microscopy, and a protease accessibility experiment. The surface-exposed fusion protein retains the MPH activity and GFP fluorescence. Anchorage of macromolecule fusions on the outer membrane neither inhibits cell growth nor affects cell viability, as shown by growth kinetics of cells and stability of resting cultures. The engineered E coli with surface-expressed MPH-GFP has two major advantages over the same strain expressing cytosolic MPH-GFP, including 7-fold higher whole-cell activity and 2-fold stronger fluorescence. Moreover, the construct pLOMG33 can potentially be applied to various bacterial species for enhancing field use. This is the first report on the presentation of GFP fusions on the cell surface by Lpp-OmpA. Our results suggest that Lpp-OmpA is a useful tool for the functional display of macromolecule passenger proteins on the cell surface.
引用
收藏
页码:6105 / 6110
页数:6
相关论文
共 50 条
  • [41] Oxidation of carbonyl compounds by whole-cell biocatalyst
    K. R. Gawai
    P. D. Lokhande
    K. M. Kodam
    I. Soojhawon
    World Journal of Microbiology and Biotechnology, 2005, 21 : 457 - 461
  • [42] A synergetic whole-cell biocatalyst for biodiesel production
    Yan, Yunjun
    Xu, Li
    Dai, Min
    RSC ADVANCES, 2012, 2 (15) : 6170 - 6173
  • [43] Oxidation of carbonyl compounds by whole-cell biocatalyst
    Gawai, KR
    Lokhande, PD
    Kodam, KM
    Soojhawon, I
    WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, 2005, 21 (04): : 457 - 461
  • [44] Production of biodiesel by immobilized whole-cell biocatalyst
    Chen, Jyh-Ping
    Chang, Shu-Chin
    JOURNAL OF BIOTECHNOLOGY, 2008, 136 : S385 - S386
  • [45] Development of a Continuous Bioconversion System Using a Thermophilic Whole-Cell Biocatalyst
    Pham Huynh Ninh
    Honda, Kohsuke
    Yokohigashi, Yukako
    Okano, Kenji
    Omasa, Takeshi
    Ohtake, Hisao
    APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2013, 79 (06) : 1996 - 2001
  • [46] Whole-cell catalysis by surface display of fluorinase on Escherichia coli using N-terminal domain of ice nucleation protein
    Xinming Feng
    Miaomiao Jin
    Wei Huang
    Wei Liu
    Mo Xian
    Microbial Cell Factories, 20
  • [47] Whole-cell catalysis by surface display of fluorinase on Escherichia coli using N-terminal domain of ice nucleation protein
    Feng, Xinming
    Jin, Miaomiao
    Huang, Wei
    Liu, Wei
    Xian, Mo
    MICROBIAL CELL FACTORIES, 2021, 20 (01)
  • [48] Biosynthesis of 4-hydroxybenzylideneacetone by Whole-Cell Escherichia coli
    Zhu, Xingmiao
    Chen, Pengcheng
    Zheng, Pu
    CATALYSTS, 2022, 12 (09)
  • [49] Construction and characterization of a recombinant whole-cell biocatalyst of Escherichia coli expressing styrene monooxygenase under the control of arabinose promoter
    Jong Wan Bae
    Seunghee Shin
    S. Mohan Raj
    Song Eun Lee
    Sun-Gu Lee
    Yong-Joo Jeong
    Sunghoon Park
    Biotechnology and Bioprocess Engineering, 2008, 13 : 69 - 76
  • [50] Whole-Cell Display of Phospholipase D in Escherichia coli for High-Efficiency Extracellular Phosphatidylserine Production
    Sun, Baotong
    Li, Zhongchen
    Peng, Yanhong
    Wang, Fei
    Cheng, Yibin
    Liu, Yang
    Ma, Lixin
    BIOMOLECULES, 2024, 14 (04)