Construction of Xylose Dehydrogenase Displayed on the Surface of Bacteria Using Ice Nucleation Protein for Sensitive d-Xylose Detection
- 13 December 2011
- journal article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 84 (1), 275-282
- https://doi.org/10.1021/ac202513u
Abstract
A novel method was developed to detect d-xylose (INS 967) sensitively and selectively, which is based on a xylose dehydrogenase (XDH) cell-surface displaying system using a newly identified ice nucleation protein from Pseudomonas borealis DL7 as an anchoring motif. With coenzyme NAD(+), the XDH-displayed bacteria facilitates the catalysis of the oxidization of xylose and the resultant NADH can be detected spectrometrically at 340 nm. The fusion protein was characterized by proteinase accessibility, Western blot, and enzyme activity assays. The established XDH surface displaying system did not inhibit the growth of the recombinant Escherichia coli strain. The XDH was mainly displayed on the surface of host cells, which is of high XDH activity and high D-xylose specificity. The optimal temperature and pH of cell displayed XDH were found at 30 °C and pH 8.0, respectively. The XDH-displayed bacteria can be used directly without further enzyme extraction and purification, and it improved the stability of the enzyme. Moreover, the cell-surface-displayed-protein-based approach showed a wide linear range (5-900 μM) and a low detection limit of 2 μM of d-xylose. More importantly, the recombinant cells could be used for precise detection of D-xylose from the real samples such as foods and degradation products of lignocellulose. The method shown here provides a simple, rapid, and low-cost strategy for the sensitive and selective measurement of D-xylose. In addition, the XDH-displayed bacteria showed an interesting response in developing electrochemical biosensors. Thus, the genetically engineered cells may find broad application in such biosensors and biocatalysts. Similarly, this type of genetic approach may be used for the expression of other intracellular enzymes of interest for certain purposes.Keywords
This publication has 42 references indexed in Scilit:
- Near Infrared Spectroscopy As High-Throughput Technology for Screening of Xylose-Fermenting Recombinant Saccharomyces cerevisiae StrainsAnalytical Chemistry, 2011
- Decorating microbes: surface display of proteins on Escherichia coliTrends in Biotechnology, 2010
- Surface Display of Heme- and Diflavin-Containing Cytochrome P450 BM3 in Escherichia coli: A Whole-Cell Biocatalyst for OxidationJournal of Microbiology and Biotechnology, 2010
- Quantification of Xylitol in Foods by an Indirect Competitive ImmunoassayJournal of Agricultural and Food Chemistry, 2010
- Cotranslocation of Methyl Parathion Hydrolase to the Periplasm and of Organophosphorus Hydrolase to the Cell Surface of Escherichia coli by the Tat Pathway and Ice Nucleation Protein Display SystemApplied and Environmental Microbiology, 2010
- d-Xylose Degradation Pathway in the Halophilic Archaeon Haloferax volcaniiJournal of Biological Chemistry, 2009
- Bioethanol production performance of five recombinant strains of laboratory and industrial xylose-fermenting Saccharomyces cerevisiaeBioresource Technology, 2009
- Genetic Analysis of a Novel Pathway for d -Xylose Metabolism in Caulobacter crescentusJournal of Bacteriology, 2007
- Cell surface display of Chi92 onEscherichia coliusing ice nucleation protein for improved catalytic and antifungal activityFEMS Microbiology Letters, 2006
- Erwinia herbicola: A Bacterial Ice Nucleus Active in Increasing Frost Injury to CornPhytopathology®, 1978