Versatile microbial surface-display for environmental remediation and biofuels production

Wu, Cindy H.; Mulchandani, Ashok; Chen, Wilfred

doi:10.1016/j.tim.2008.01.003

Cited by 101 publications

(66 citation statements)

References 80 publications

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“…Other uses include the display of enzymes (35) as whole-cell biocatalysts, display of binding molecules such as antibodies or affibodies (35) (in diagnostics as biosensors, in therapy for pathogen or toxin removal, and in bioremediation for heavy metal binding [36]), and display of proteins for protein engineering by using combinatorial libraries and in vitro selection (8).…”

mentioning

confidence: 99%

Identification of Candidate Carrier Proteins for Surface Display on Lactococcus lactis by Theoretical and Experimental Analyses of the Surface Proteome

Berlec

Zadravec

Jevnikar

et al. 2011

Appl Environ Microbiol

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Lactococcus lactis is a lactic acid bacterium of proven safety for use in human oral applications. For this purpose, surface display of recombinant proteins is important, and new approaches for it are being sought. Analysis of the bacterial surface proteome is essential in identifying new candidate carrier proteins for surface display. We have made two different predictions of surface-associated proteins of L. lactis MG1363 by using Augur and LocateP software, which yielded 666 and 648 proteins, respectively. Surface proteins of L. lactis NZ9000, a derivative of MG1363, were identified by using a proteomics approach. The surface proteins were cleaved from intact bacteria, and the resulting peptides were identified by mass spectrometry. The latter approach yielded 80 proteins, 34 of which were not predicted by either software. Of the 80 proteins, 7 were selected for further study. These were cloned in frame with a C-terminal hexahistidine tag and overexpressed in L. lactis NZ9000 using nisin-controlled expression. Proteins of correct molecular weight carrying a hexahistidine tag were detected. Their surface localization was confirmed with flow cytometry. Basic membrane protein A (BmpA) was exposed at the highest level. To test BmpA as a candidate carrier protein, the hexahistidine tag was replaced by the B domain of staphylococcal protein A in the genetic construct. The B domain was displayed on the surface with BmpA as a carrier. The advantage of covalent BmpA binding was demonstrated. BmpA was thus shown to be a suitable candidate for a carrier protein in lactococcal surface display.Lactic acid bacteria (LAB) are among the most intensively exploited microorganisms in the dairy industry. They are used in the fermentation of foodstuffs and as probiotics on account of their health benefits (11). Lactococcus lactis is a model LAB with industrially important applications. The genomes of several L. lactis strains have been sequenced, but strains MG1363 (33) and IL1403 (4) are most commonly used in laboratories.Various biotechnological applications of surface display of recombinant proteins in bacteria have been suggested, with vaccine delivery being the most popular in LAB (20,34). Other uses include the display of enzymes (35) as whole-cell biocatalysts, display of binding molecules such as antibodies or affibodies (35) (in diagnostics as biosensors, in therapy for pathogen or toxin removal, and in bioremediation for heavy metal binding [36]), and display of proteins for protein engineering by using combinatorial libraries and in vitro selection (8).The majority of surface display techniques have been developed for Gram-negative bacteria, with autodisplay being probably the most efficacious technique (16). However, several other approaches for surface display in Gram-positive bacteria have also been described. Transmembrane proteins, lipoproteins, LPXTG-like proteins, and cell wall-binding proteins have been displayed on the surfaces of Gram-positive bacteria (10). Two of these display types have been commonly explo...

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confidence: 99%

Identification of Candidate Carrier Proteins for Surface Display on Lactococcus lactis by Theoretical and Experimental Analyses of the Surface Proteome

Berlec

Zadravec

Jevnikar

et al. 2011

Appl Environ Microbiol

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“…Yeast and bacterial cell systems where proteins or peptides are expected on the cell exterior wer e r e p o r t e d t o b e u s e d a s b i o c a t a l y s t s , biosorbents and biostimulants (Wu et al, 2008).…”

Section: Protein Engineering 44mentioning

confidence: 99%

Protein Engineering Methods and Applications

Turanlı-Yıldız¹,

Alkım²,

Petek³

2012

Protein Engineering

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“…1 The yeast Saccharomyces cerevisiae is particularly suited for these applications because it has a rigid cell wall, and heterologous proteins can be efficiently targeted to the glycoprotein layer. For example, heterologous proteins have been fused to the glycosylphosphatidylinositol (GPI)-anchor signaling domain of yeast mannoprotein, and the fusion products were expressed as yeast cell-surface proteins.…”

Section: Introductionmentioning

confidence: 99%

Estimation of Enzyme Kinetic Parameters of Cell Surface-displayed Organophosphorus Hydrolase and Construction of a Biosensing System for Organophosphorus Compounds

et al. 2011

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Versatile microbial surface-display for environmental remediation and biofuels production

Cited by 101 publications

References 80 publications

Identification of Candidate Carrier Proteins for Surface Display on Lactococcus lactis by Theoretical and Experimental Analyses of the Surface Proteome

Identification of Candidate Carrier Proteins for Surface Display on Lactococcus lactis by Theoretical and Experimental Analyses of the Surface Proteome

Protein Engineering Methods and Applications

Estimation of Enzyme Kinetic Parameters of Cell Surface-displayed Organophosphorus Hydrolase and Construction of a Biosensing System for Organophosphorus Compounds

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