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

Wee, Joanne Mei Yuin and Abd. Murad, Abdul Munir and Abu Bakar, Farah Diba and Low, Kheng Oon and Md. Illias, Rosli (2019) Expression of xylanase on Escherichia coli using a truncated ice nucleation protein of Erwinia ananas (InaA). Process Biochemistry, 78 . pp. 25-32. ISSN 1359-5113

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Official URL: http://dx.doi.org/10.1016/j.procbio.2019.01.005

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.

Item Type:Article
Uncontrolled Keywords:ice nucleation protein, surface display, xylanase
Subjects:T Technology > TP Chemical technology
Divisions:Chemical and Energy Engineering
ID Code:88672
Deposited By: Yanti Mohd Shah
Deposited On:15 Dec 2020 18:53
Last Modified:15 Dec 2020 18:53

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