Expression of Cell Wall-Modifying Enzymes in Aspen for Improved Lignocellulose Processing

Abstract

Wood is an important source of biomass for materials and chemicals, and a target for genetic engineering of its properties for different applications or for research. Wood properties can be altered by using different enzymes acting on cell wall polymers postsynthetically in cell walls. This approach allows for a precise polymer structure modification thanks to the specificity of enzymes used. Such enzymes can originate from all kinds of organisms, or even be modified in a desired way for novel attributes. Here we present a general strategy for expressing a microbial enzyme in aspen and targeting it to cell wall, using an example of fungal glucuronoyl esterase. We describe methods of vector cloning, plant transformation, transgenic line selection and multiplication, testing for the presence of enzymatic activity in different cell compartments, and finally the method of plant transferring from sterile culture to the greenhouse conditions.

Keywords

Fungal wood degrading enzymes; Protein targeting to cell wall; Gateway (R) cloning; Aspen transformation; Transgenic trees; Populus tremula x tremuloides; Vector design; In vitro culture

Published in

Methods in Molecular Biology
2020, Volume: 2149, pages: 145-164 Title: The Plant Cell Wall : Methods and Protocols
ISBN: 978-1-0716-0619-3, eISBN: 978-1-0716-0621-6Publisher: HUMANA PRESS INC, 999 RIVERVIEW DR, STE 208, TOTOWA, NJ 07512-1165 USA

SLU Authors

• Derba-Maceluch, Marta

  • Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences
    

• Mellerowicz, Ewa

  • Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Wood Science


Publication identifier

DOI: https://doi.org/10.1007/978-1-0716-0621-6_9

Permanent link to this page (URI)

https://res.slu.se/id/publ/113307