Cell Surface Display of Functional Macromolecule Fusions on Escherichia coli for Development of an Autofluorescent Whole-Cell Biocatalyst

In this research, xylan was utilized by a recombinant whole cell biocatalyst, which was developed by expressing three xylanases — β-xylosidase, endoxylanase, and α-arabinofuranosidase — on the surface of the E. coli BL21 (DE3). The xylanases were displayed on the surface of the cells by fusing with anchor proteins, Blc. The assimilation of xylan by cell surface display was the first step in the consolidated bioprocessing (CBP). This result shows that the engineering strains could be endowed with the ability to assimilate xylan. The co-display engineering strains utilized xylan and expressed less metabolic burden than the engineering strains secreting extracellular xylanases.

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Cell Surface Display of Yarrowia lipolytica Lipase Lip2p Using the Cell Wall Protein YlPir1p, Its Characterization, and Application as a Whole-Cell Biocatalyst

Applied Biochemistry and Biotechnology ◽

10.1007/s12010-015-1557-7 ◽

2015 ◽

Vol 175 (8) ◽

pp. 3888-3900 ◽

Cited By ~ 18

Author(s):

Evgeniya Y. Yuzbasheva ◽

Tigran V. Yuzbashev ◽

Natalia I. Perkovskaya ◽

Elizaveta B. Mostova ◽

Tatiana V. Vybornaya ◽

...

Keyword(s):

Cell Wall ◽

Cell Surface ◽

Yarrowia Lipolytica ◽

Surface Display ◽

Cell Surface Display ◽

Cell Wall Protein ◽

Whole Cell ◽

Whole Cell Biocatalyst

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Research on Construction of a Whole-Cell Biocatalyst for Xylan Degradation

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.347-353.2599 ◽

2011 ◽

Vol 347-353 ◽

pp. 2599-2603

Author(s):

Jian Zhang Lu ◽

Mei Lin Cui ◽

Shan Shan Du ◽

Lu Yang ◽

Qin Guo ◽

...

Keyword(s):

Cell Surface ◽

Surface Display ◽

Cell Surface Display ◽

Whole Cell ◽

Secretion Signal ◽

Gal1 Promoter ◽

Dry Cell Weight ◽

Whole Cell Biocatalyst ◽

Genes Encoding ◽

Secretion Expression

Endo-1,4-β-xylanase (E.C.3.2.1.8) is a family of glycoside hydrolase. It is capable of hydrolyzing the backbone of substituted xylan polymers into fragments of random size. Due to this ability, xylanase can serve the degradation of lignocellulose, and facilitate the application of xylan. Cell-surface display of enzymes is one of the most attractive applications in yeast. It is a promising utilization in constructing the whole-cell biocatalyst of xylanase. For this purpose, a cDNA sequence of endo-1,4-β-xylanase B (XylB) from Aspergillus niger BCC14405 was optimized and synthesized according to the codon bias of Saccharomyces cerevisiae. The genes encoding galactokinase (GAL1) promoter, α-mating factor 1 (MFα1) pre-pro secretion signal, fully codon-optimized XylB, the 320 amino acids of C terminal of α-agglutinin, alcohol dehydrogenase (ADH1) terminator and kanMX cassette were amplified and cloned into YEplac181 to construct a cell-surface display vector called pGMAAK-XylB with α-agglutinin as an anchor. Then pGMAAK-XylB was transformed into S. cerevisiae. The results show XylB was immobilized and actively expressed on S. cerevisiae. Meanwhile, a secretion expression plasmid was also constructed using the above elements except α-agglutinin as a control strain in the study of characteristic of XylB. After an induction of 48 h by 2% galactose, the activity of displayed XylB reached 63 U/g dry-cell weight. The optimal pH of displayed XylB has changed from 5 to 6 and the optimal temperature has changed from 50 °C to 60 °C, comparing to the recombinant secretion XylB.

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