The Suitability of Orthogonal Hosts to Study Plant Cell Wall Biosynthesis

Abstract

Plant cells are surrounded by an extracellular matrix that consists mainly of polysaccharides. Many molecular components involved in plant cell wall polymer synthesis have been identified, but it remains largely unknown how these molecular players function together to define the length and decoration pattern of a polysaccharide. Synthetic biology can be applied to answer questions beyond individual glycosyltransferases by reconstructing entire biosynthetic machineries required to produce a complete wall polysaccharide. Recently, this approach was successful in establishing the production of heteromannan from several plant species in an orthogonal host—a yeast—illuminating the role of an auxiliary protein in the biosynthetic process. In this review we evaluate to what extent a selection of organisms from three kingdoms of life (Bacteria, Fungi and Animalia) might be suitable for the synthesis of plant cell wall polysaccharides. By identifying their key attributes for glycoengineering as well as analyzing the glycosidic linkages of their native polymers, we present a valuable comparison of their key advantages and limitations for the production of different classes of plant polysaccharides.

Modular biosynthesis of plant hemicellulose and its impact on yeast cells

“…Of these five samples, only AkCSLA3, 2232 and 3332 also increased 4-Glc to ≥4.5% (Fig. 3b) from the small amounts (1.6–3.3%) that are natively part of yeast compounds such as glycogen [3]. Glucomannan production by AkCSLA3 and 3332 was supported by both glycosidic linkage (Fig.…”

“…Synthetic biology efforts in this area have been largely restricted to bacterial cellulose due to its well-characterized production [1,2]. Several microbial organisms have been identified as suitable hosts for orthogonal polysaccharide production [3]. Chief among them is the industrial yeast Pichia pastoris , which was recently engineered into an autotroph capable of growing on the greenhouse gas CO 2 [4].…”

“…We optimized the cultivation of Pichia pastoris for the orthogonal production of plant hemicelluloses, and enhanced HM biosynthesis by assembling modular CSLA enzymes. Pichia cells were promising hosts to study CSLC and CSLA activities [3,18,24,25], but the impact of the plant polymers was not previously investigated. Here, we extend their utility by improving the speed of chimeric enzyme expression and the hemicellulose yields.…”

Modular biosynthesis of plant hemicellulose and its impact on yeast cells

“…Synthetic biology efforts in this area have been largely restricted to bacterial cellulose due to its well-characterized production [ 1 , 2 ]. Several microbial organisms have been identified as suitable hosts for orthogonal polysaccharide production [ 3 ]. Chief among them is the industrial yeast Pichia pastoris , which was recently engineered into an autotroph capable of growing on the greenhouse gas CO 2 [ 4 ].…”

“…We optimized the cultivation of Pichia pastoris for the orthogonal production of plant hemicelluloses, and enhanced HM biosynthesis by assembling modular CSLA enzymes. Pichia cells were promising hosts to study CSLC and CSLA activities [ 3 , 18 , 24 , 25 ], but the impact of the plant polymers was not previously investigated. Here, we extend their utility by improving the speed of chimeric enzyme expression and the hemicellulose yields.…”