ECI Digital Archives

Title

Critical role of metals in biochemical properties of xylose isomerase

Authors

Misun Lee, University of Groningen, NetherlandsFollow
Dick B. Janssen, University of Groningen, Netherlands
Henriette J. Rozeboom, University of Groningen, Netherlands
Maarten D. Verhoeven, Delft University of Technology, The Netherlands
Jack T. Pronk, Delft University of Technology, The Netherlands
Paul de Waal, DSM Biotechnology Center, Alexander Fleminglaan 1, 2613 AX Delft, The Netherlands

Conference Dates

September 24-28, 2017

Abstract

Improving the activity of xylose isomerase (XI) is highly desired for achieving efficient fermentation of xylose in lignocellulosic biomass using XI-expressing S. cerevisiae. XI is a metalloenzyme which requires two bivalent metals for its catalytic activity. The enzyme from Piromyces sp. E2 (PirXI)^[1],[2] is activated with various metal ions including Mg²⁺, Mn²⁺, Ca²⁺, Co²⁺, Zn²⁺ and Fe²⁺. The biochemical properties of PirXI are dependent on the types of its metal cofactors. Moreover, the enzyme shows different affinities towards these metals. Characterization of these properties is critical for understanding the enzyme behavior in vivo and to further adapt the enzyme to the cytosolic metal environment. Recently, we have shown that altered intracellular metal composition can improve anaerobic growth of a xylose-fermenting strain by enhancing the activity of PirXI^[3]. Furthermore, our current study on PirXI and other studies on different XIs have shown that it is also possible to change the metal preferences of the enzyme^[4]. A PirXI variant with a single amino acid substitution in the proximity of the metal binding residues showed significant changes in metal preference compared to the wild-type PirXI. Further exploration on metal specificity of PirXI is necessary to optimize the in vivo enzyme activity.

References:

1.Kuyper M, Harhangi HR, Stave AK, Winkler AA, Jetten MS, de Laat WT, den Ridder JJ, Op den Camp HJ, van Dijken JP, Pronk JT, FEMS Yeast Research 4 (2003) 69-78

2.van Maris AJ, Winkler AA, Kuyper M, de Laat WT, van Dijken JP, Pronk JT, Adv Biochem Engin/Biotechnol (2007) 108: 179–204

3.Verhoeven MD, Lee M, Kamoen L, van den Broek M, Janssen DB, Daran JG, van Maris AJA, Pronk JT, Sci. Rep (2017) 7, 46155

4.van Tilbeurgh H, Jenkins J, Chiadmi M, Janin J, Wodak SJ, Mrabet NT, Lambeir AM, Biochemistry (1992) 31: 5467-5471

Recommended Citation

Misun Lee, Dick B. Janssen, Henriette J. Rozeboom, Maarten D. Verhoeven, Jack T. Pronk, and Paul de Waal, "Critical role of metals in biochemical properties of xylose isomerase" in "Enzyme Engineering XXIV", Pierre Monsan, Toulouse White Biotechnology, France Magali Remaud-Simeon, LISBP-INSA, University of Toulouse, France Eds, ECI Symposium Series, (2017). https://dc.engconfintl.org/enzyme_xxiv/43