Expression of xylanase on Escherichia coli using a truncated ice nucleation protein of Erwinia ananas (InaA)

Mei Yuin Joanne Wee, Abdul Munir Abd. Murad, Farah Diba Abu Bakar, Kheng Oon Low, Rosli Md Illias

Research output: Contribution to journalArticle

Abstract

Cell surface display involves the anchoring of enzymes on the surface of cells to be used as whole-cell biocatalysts. In this study, an ice nucleation protein (INP) anchor from Erwinia ananas IN-10, InaA, was used to display xylanase. The ability of InaA to be targeted to the outer membrane was compared to cells displaying xylanase using two other INPs, InaK and InaZ. SDS-PAGE and western blot of the outer membrane fraction proved that surface targeting was successful. Whole-cell xylanase activity showed that InaA anchored xylanase gave an activity of 92.2 U/g dry cell weight which was up to three times higher than the other two display constructs used. Surface anchoring of InaA was up to four times better compared to the other two INP anchors as was confirmed by flow cytometry. Expression of InaA on the surface was optimized by one-factor-at-a-time (OFAT) to obtain the optimum parameter conditions for highest surface expression. The results presented showed that InaA is an excellent INP for surface display for xylanase and has great potential in the degradation of xylan.

Original languageEnglish
JournalProcess Biochemistry
DOIs
Publication statusAccepted/In press - 1 Jan 2019

Fingerprint

Erwinia
Ananas
Escherichia coli
Ice
Nucleation
Proteins
Display devices
Anchors
Xylans
Membranes
Enzymes
Biocatalysts
Flow cytometry
ice nucleation protein
Polyacrylamide Gel Electrophoresis
Flow Cytometry
Western Blotting
Weights and Measures
Cells
Degradation

Keywords

  • Ice nucleation protein
  • Surface display
  • Xylanase

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

Cite this

Expression of xylanase on Escherichia coli using a truncated ice nucleation protein of Erwinia ananas (InaA). / Wee, Mei Yuin Joanne; Abd. Murad, Abdul Munir; Abu Bakar, Farah Diba; Low, Kheng Oon; Md Illias, Rosli.

In: Process Biochemistry, 01.01.2019.

Research output: Contribution to journalArticle

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AU - Low, Kheng Oon

AU - Md Illias, Rosli

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