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- Surface Display Expression of Bacillus licheniformis Lipase in Escherichia coli Using Lpp’OmpA Chimera
- Jae-Hyung Jo , Chan-Wook Han , Seung-Hwan Kim , Hyuk-Jin Kwon , Hyune-Hwan Lee
- J. Microbiol. 2014;52(10):856-862. Published online August 27, 2014
- DOI: https://doi.org/10.1007/s12275-014-4217-7
- 46 View
- 0 Download
- 15 Crossref
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Abstract
- The lipase from Bacillus licheniformis ATCC14580 was displayed on the cell surface of Escherichia coli using Lpp’OmpA as the anchoring protein. The expressed Lpp’OmpA-lipase fusion protein has a molecular weight of approximately 35 kDa, which was confirmed by SDS-PAGE and western blot analysis. The Lpp’OmpA-lipase fusion protein was located on the cell surface, as determined by immunofluorescence confocal microscopy and flow cytometry. The enzyme activity of the surface-displayed lipase showed clear halo around the colony. The cell surface-displayed lipase showed the highest activity of 248.12 ± 9.42 U/g (lyophilized cell) at the optimal temperature of 37°C and pH 8.0. The enzyme exhibited the highest activity toward the substrate p-nitrophenyl caprylate (C8). These results suggest that E. coli, which displayed the lipase on its surface, could be used as a whole cell biocatalyst.
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Citations
Citations to this article as recorded by
- Recent advances in bioinspired multienzyme engineering for food applications
Xianhan Chen, Yujin Chen, Dandan Tang, Mengyu Li, Yuting Lu, Yi Cao, Quanyu Zhao, Shuai Jiang, Wei Liu, Ling Jiang
Trends in Food Science & Technology.2025; 156: 104840. CrossRef - Surface Engineering of Escherichia coli to Display Its Phytase (AppA) and Functional Analysis of Enzyme Activities
Patricia L. A. Muñoz-Muñoz, Celina Terán-Ramírez, Rosa E. Mares-Alejandre, Ariana B. Márquez-González, Pablo A. Madero-Ayala, Samuel G. Meléndez-López, Marco A. Ramos-Ibarra
Current Issues in Molecular Biology.2024; 46(4): 3424. CrossRef - Characterization of a novel subfamily 1.4 lipase from Bacillus licheniformis IBRL-CHS2: Cloning and expression optimization
Ammar Khazaal Kadhim Almansoori, Nidyaletchmy Subba Reddy, Mustafa Abdulfattah, Sarah Solehah Ismail, Rashidah Abdul Rahim, Estibaliz Sansinenea
PLOS ONE.2024; 19(12): e0314556. CrossRef - Surface Display of Multiple Metal-Binding Domains in Deinococcus radiodurans Alleviates Cadmium and Lead Toxicity in Rice
Liangyan Wang, Yudong Wang, Shang Dai, Binqiang Wang
International Journal of Molecular Sciences.2024; 25(23): 12570. CrossRef - A bacterial outer membrane vesicle-based click vaccine elicits potent immune response against Staphylococcus aureus in mice
Jingjing Sun, Xuansheng Lin, Yige He, Baozhong Zhang, Nan Zhou, Jian-dong Huang
Frontiers in Immunology.2023;[Epub] CrossRef - Establishment of a soluble expression and rapid purification system for self-assembling protein nanoparticle and characterization of its physiochemical properties
Dan Wang, Linwei Duan, Min Wei, Baizhu Chen, Zhipeng Li, Qingyou Liu
Biochemical Engineering Journal.2022; 186: 108580. CrossRef - A Modular System for the Rapid Comparison of Different Membrane Anchors for Surface Display on Escherichia coli
Sabrina Gallus, Esther Mittmann, Kersten S. Rabe
ChemBioChem.2022;[Epub] CrossRef - Decorating the surface of Escherichia coli with bacterial lipoproteins: a comparative analysis of different display systems
Sonia Nicchi, Maria Giuliani, Fabiola Giusti, Laura Pancotto, Domenico Maione, Isabel Delany, Cesira L. Galeotti, Cecilia Brettoni
Microbial Cell Factories.2021;[Epub] CrossRef - Recombinant expression and surface display of a zearalenone lactonohydrolase from Trichoderma aggressivum in Escherichia coli
Shurong Chen, Li Pan, Siying Liu, Lijie Pan, Xuejie Li, Bin Wang
Protein Expression and Purification.2021; 187: 105933. CrossRef - Bacterial Cell Display as a Robust and Versatile Platform for Engineering Low‐Affinity Ligands and Enzymes
Eszter Csibra, Marleen Renders, Vitor B. Pinheiro
ChemBioChem.2020; 21(19): 2844. CrossRef - Surface Display of Complex Enzymes by in Situ SpyCatcher‐SpyTag Interaction
Sabrina Gallus, Theo Peschke, Malte Paulsen, Teresa Burgahn, Christof M. Niemeyer, Kersten S. Rabe
ChemBioChem.2020; 21(15): 2126. CrossRef - Shaking Rate during Production Affects the Activity of Escherichia coli Surface-Displayed Candida antarctica Lipase A
Chen-Fu Chung, Shih-Che Lin, Tzong-Yuan Juang, Yung-Chuan Liu
Catalysts.2020; 10(4): 382. CrossRef - Functional Display of an Amoebic Chitinase in Escherichia coli Expressing the Catalytic Domain of EhCHT1 on the Bacterial Cell Surface
Ricardo Torres-Bañaga, Rosa E. Mares-Alejandre, Celina Terán-Ramírez, Ana L. Estrada-González, Patricia L.A. Muñoz-Muñoz, Samuel G. Meléndez-López, Ignacio A. Rivero, Marco A. Ramos-Ibarra
Applied Biochemistry and Biotechnology.2020; 192(4): 1255. CrossRef - Heterologous expression of antigenic peptides in Bacillus subtilis biofilms
Cédric M. Vogt, Elisabeth M. Schraner, Claudio Aguilar, Catherine Eichwald
Microbial Cell Factories.2016;[Epub] CrossRef - Display of Fungi Xylanase on Escherichia coli Cell Surface and Use of the Enzyme in Xylan Biodegradation
Wei Qu, Yuanxia Xue, Qiang Ding
Current Microbiology.2015; 70(6): 779. CrossRef
- Recent advances in bioinspired multienzyme engineering for food applications
- Cell-Surface Expression of Aspergillus saitoi-Derived Functional α-1,2-Mannosidase on Yarrowia lipolytica for Glycan Remodeling
- Hye Yun Moon , Trinh Luu Van , Seon Ah Cheon , Jinho Choo , Jeong-Yoon Kim , Hyun Ah Kang
- J. Microbiol. 2013;51(4):506-514. Published online August 30, 2013
- DOI: https://doi.org/10.1007/s12275-013-3344-x
- 40 View
- 0 Download
- 13 Crossref
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Abstract
- Expression of proteins on the surface of yeast has a wide range of applications, such as development of live vaccines, screening of antibody libraries, and use as whole-cell biocatalysts. The hemiascomycetes yeast Yarrowia lipolytica has been raised as a potential host for heterologous expression of recombinant proteins. In this study, we report the expression of Aspergillus saitoi α-1,2-mannosidase, encoded by the msdS gene, on the cell surface of Y. lipolytica. As the first step to achieve the secretory expression of msdS protein, four different signal sequences-derived from the endogenous Y. lipolytica Lip2 and Xpr2 prepro regions and the heterologous A. niger α-amylase and rice α-amylase signal sequences-were analyzed for their secretion efficiency. It was shown that the YlLip2 prepro sequence was most efficient in directing the secretory expression of msdS in fully N-glycosylated forms. The surface display of msdS was subsequently directed by fusing GPI anchoring motifs derived from Y. lipolytica cell wall proteins, YlCwp1p and YlYwp1p, respectively, to the C-terminus of the Lip2 prepro-msdS protein. The expression of actively functional msdS protein on the cell surface was confirmed by western blot, flow cytometry analysis, along with the α-1,2-mannosidase activity assay using intact Y. lipolytica cells as the enzyme source. Furthermore, the glycoengineered Y. lipolytica Δoch1Δmpo1 strains displaying α-1,2-mannosidase were able to convert Man8GlcNAc2 to Man5GlcNAc2 efficiently on their cell-wall mannoproteins, demonstrating its potential used for glycoengineering in vitro or in vivo.
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Citations
Citations to this article as recorded by- Engineering novel Yarrowia lipolytica whole-cell biocatalysts by cell surface display of the native Lip2 lipase for biodiesel production
Maria Orfanidou, Eleftheria Panagiotidou, Antonios M. Makris, Eleni Theodosiou
Biotechnology for the Environment.2025;[Epub] CrossRef - Yeast Surface Display System: Strategies for Improvement and Biotechnological Applications
Karla V. Teymennet-Ramírez, Fernando Martínez-Morales, María R. Trejo-Hernández
Frontiers in Bioengineering and Biotechnology.2022;[Epub] CrossRef - Dietary intake of GDF11 delays the onset of several biomarkers of aging in male mice through anti-oxidant system via Smad2/3 pathway
Lili Song, Fei Wu, Congjun Li, Shicui Zhang
Biogerontology.2022; 23(3): 341. CrossRef - Hydrolytic secretome engineering in Yarrowia lipolytica for consolidated bioprocessing on polysaccharide resources: review on starch, cellulose, xylan, and inulin
Ewelina Celińska, Jean-Marc Nicaud, Wojciech Białas
Applied Microbiology and Biotechnology.2021; 105(3): 975. CrossRef - Yarrowia lipolytica Strains and Their Biotechnological Applications: How Natural Biodiversity and Metabolic Engineering Could Contribute to Cell Factories Improvement
Catherine Madzak
Journal of Fungi.2021; 7(7): 548. CrossRef - Engineering Yarrowia lipolytica for Use in Biotechnological Applications: A Review of Major Achievements and Recent Innovations
Catherine Madzak
Molecular Biotechnology.2018; 60(8): 621. CrossRef - Synthetic biology tools for engineering Yarrowia lipolytica
M. Larroude, T. Rossignol, J.-M. Nicaud, R. Ledesma-Amaro
Biotechnology Advances.2018; 36(8): 2150. CrossRef - Development of recombinant Yarrowia lipolytica producing virus-like particles of a fish nervous necrosis virus
Van-Trinh Luu, Hye Yun Moon, Jee Youn Hwang, Bo-Kyu Kang, Hyun Ah Kang
Journal of Microbiology.2017; 55(8): 655. CrossRef - Using a vector pool containing variable-strength promoters to optimize protein production in Yarrowia lipolytica
Rémi Dulermo, François Brunel, Thierry Dulermo, Rodrigo Ledesma-Amaro, Jérémy Vion, Marion Trassaert, Stéphane Thomas, Jean-Marc Nicaud, Christophe Leplat
Microbial Cell Factories.2017;[Epub] CrossRef -
Yarrowia lipolytica: recent achievements in heterologous protein expression and pathway engineering
Catherine Madzak
Applied Microbiology and Biotechnology.2015; 99(11): 4559. CrossRef - Biotechnological applications of Yarrowia lipolytica: Past, present and future
Hu-Hu Liu, Xiao-Jun Ji, He Huang
Biotechnology Advances.2015; 33(8): 1522. CrossRef - Functional characterization of extracellular chitinase encoded by the YlCTS1 gene in a dimorphic yeast Yarrowia lipolytica
Jeong-Nam Park, Chang Pyo Han, Dong-Jik Lee, Seon Ah Cheon, Hyun Ah Kang
Journal of Microbiology.2014; 52(4): 284. CrossRef - Characterization of putative glycosylphosphatidylinositol-anchoring motifs for surface display in the methylotrophic yeast Hansenula polymorpha
Seon Ah Cheon, Jinhee Jung, Jin Ho Choo, Doo-Byoung Oh, Hyun Ah Kang
Biotechnology Letters.2014; 36(10): 2085. CrossRef
- Engineering novel Yarrowia lipolytica whole-cell biocatalysts by cell surface display of the native Lip2 lipase for biodiesel production
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