Studies on the structure and function cellulases and hemicellulases.

Author(s):  
H.J. Gilbert ◽  
G.P. Hazlewood

Plant structural polysaccharides provide a major source of nutrient for ruminant livestock. These carbohydrates are not degraded by mammalian-derived enzymes, but are hydrolysed by rumen microbial plant cell wall hydrolases. In view of the pivotal role of microbial cellulases and xylanases in ruminant nutrition, there has been considerable interest in these enzymes. In this paper we wish to illustrate how recombinant DNA (rDNA) technology can be utilised to dissect the biochemistry and molecular architecture of these enzymes, and provides us with the opportunity of generating novel cellulases and xylanases with increased capacity to hydrolyse the plant cell wall.In general cellulases and xylanases from anaerobic microbes associate into large molecular weight complexes, whose integral structures are responsible for the efficient hydrolysis of plant structural polysaccharides. The feasibility of altering these complexes, or transferring them to other organisms represents a significant challenge.

2016 ◽  
Vol 44 (1) ◽  
pp. 74-78 ◽  
Author(s):  
Marta Busse-Wicher ◽  
Nicholas J. Grantham ◽  
Jan J. Lyczakowski ◽  
Nino Nikolovski ◽  
Paul Dupree

The molecular architecture of plant secondary cell walls is still not resolved. There are several proposed structures for cellulose fibrils, the main component of plant cell walls and the conformation of other molecules is even less well known. Glucuronic acid (GlcA) substitution of xylan (GUX) enzymes, in CAZy family glycosyl transferase (GT)8, decorate the xylan backbone with various specific patterns of GlcA. It was recently discovered that dicot xylan has a domain with the side chain decorations distributed on every second unit of the backbone (xylose). If the xylan backbone folds in a similar way to glucan chains in cellulose (2-fold helix), this kind of arrangement may allow the undecorated side of the xylan chain to hydrogen bond with the hydrophilic surface of cellulose microfibrils. MD simulations suggest that such interactions are energetically stable. We discuss the possible role of this xylan decoration pattern in building of the plant cell wall.


1991 ◽  
Vol 279 (2) ◽  
pp. 343-350 ◽  
Author(s):  
J Nari ◽  
G Noat ◽  
J Ricard

The hydrolysis of p-nitrophenyl acetate catalysed by pectin methylesterase is competitively inhibited by pectin and does not require metal ions to occur. The results suggest that the activastion by metal ions may be explained by assuming that they interact with the substrate rather than with the enzyme. With pectin used as substrate, metal ions are required in order to allow the hydrolysis to occur in the presence of pectin methylesterase. This is explained by the existence of ‘blocks’ of carboxy groups on pectin that may trap enzyme molecules and thus prevent the enzyme reaction occurring. Metal ions may interact with these negatively charged groups, thus allowing the enzyme to interact with the ester bonds to be cleaved. At high concentrations, however, metal ions inhibit the enzyme reaction. This is again understandable on the basis of the view that some carboxy groups must be adjacent to the ester bond to be cleaved in order to allow the reaction to proceed. Indeed, if these groups are blocked by metal ions, the enzyme reaction cannot occur, and this is the reason for the apparent inhibition of the reaction by high concentrations of metal ions. Methylene Blue, which may be bound to pectin, may replace metal ions in the ‘activation’ and ‘inhibition’ of the enzyme reaction. A kinetic model based on these results has been proposed and fits the kinetic data very well. All the available results favour the view that metal ions do not affect the reaction through a direct interaction with enzyme, but rather with pectin.


2020 ◽  
Vol 50 (2) ◽  
pp. 176-186
Author(s):  
Yi MAN ◽  
RuiLi LI ◽  
YuFen BU ◽  
Na SUN ◽  
YanPing JING ◽  
...  

Author(s):  
Samir Medjekal ◽  
Mouloud Ghadbane

Sheep have a gastrointestinal tract similar to that of other ruminants. Their stomach is made up of four digestive organs: the rumen, the reticulum, the omasum and the abomasum. The rumen plays a role in storing ingested foods, which are fermented by a complex anaerobic rumen microbiota population with different types of interactions, positive or negative, that can occur between their microbial populations. Sheep feeding is largely based on the use of natural or cultivated fodder, which is exploited in green by grazing during the growth period of the grass and in the form of fodder preserved during the winter period. Ruminant foods are essentially of plant origin, and their constituents belong to two types of structures: intracellular constituents and cell wall components. Cellular carbohydrates play a role of metabolites or energy reserves; soluble carbohydrates account for less than 10% dry matter (DM) of foods. The plant cell wall is multi-layered and consists of primary wall and secondary wall. Fundamentally, the walls are deposited at an early stage of growth. A central blade forms the common boundary layer between two adjacent cells and occupies the location of the cell plate. Most of the plant cell walls consist of polysaccharides (cellulose, hemicellulose and pectic substances) and lignin, these constituents being highly polymerized, as well as proteins and tannins.


2011 ◽  
Vol 43 (7) ◽  
pp. 1544-1552 ◽  
Author(s):  
Gaylord Erwan Machinet ◽  
Isabelle Bertrand ◽  
Yves Barrière ◽  
Brigitte Chabbert ◽  
Sylvie Recous

2020 ◽  
Vol 117 (11) ◽  
pp. 6003-6013 ◽  
Author(s):  
Vincent W. Wu ◽  
Nils Thieme ◽  
Lori B. Huberman ◽  
Axel Dietschmann ◽  
David J. Kowbel ◽  
...  

Filamentous fungi, such asNeurospora crassa, are very efficient in deconstructing plant biomass by the secretion of an arsenal of plant cell wall-degrading enzymes, by remodeling metabolism to accommodate production of secreted enzymes, and by enabling transport and intracellular utilization of plant biomass components. Although a number of enzymes and transcriptional regulators involved in plant biomass utilization have been identified, how filamentous fungi sense and integrate nutritional information encoded in the plant cell wall into a regulatory hierarchy for optimal utilization of complex carbon sources is not understood. Here, we performed transcriptional profiling ofN. crassaon 40 different carbon sources, including plant biomass, to provide data on how fungi sense simple to complex carbohydrates. From these data, we identified regulatory factors inN. crassaand characterized one (PDR-2) associated with pectin utilization and one with pectin/hemicellulose utilization (ARA-1). Using in vitro DNA affinity purification sequencing (DAP-seq), we identified direct targets of transcription factors involved in regulating genes encoding plant cell wall-degrading enzymes. In particular, our data clarified the role of the transcription factor VIB-1 in the regulation of genes encoding plant cell wall-degrading enzymes and nutrient scavenging and revealed a major role of the carbon catabolite repressor CRE-1 in regulating the expression of major facilitator transporter genes. These data contribute to a more complete understanding of cross talk between transcription factors and their target genes, which are involved in regulating nutrient sensing and plant biomass utilization on a global level.


2020 ◽  
Vol 11 ◽  
Author(s):  
Yuan Zhang ◽  
Allan M. Showalter

For the past 5 years, clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) technology has appeared in the molecular biology research spotlight. As a game-changing player in genome editing, CRISPR/Cas9 technology has revolutionized animal research, including medical research and human gene therapy as well as plant science research, particularly for crop improvement. One of the most common applications of CRISPR/Cas9 is to generate genetic knock-out mutants. Recently, several multiplex genome editing approaches utilizing CRISPR/Cas9 were developed and applied in various aspects of plant research. Here we summarize these approaches as they relate to plants, particularly with respect to understanding the biosynthesis and function of the plant cell wall. The plant cell wall is a polysaccharide-rich cell structure that is vital to plant cell formation, growth, and development. Humans are heavily dependent on the byproducts of the plant cell wall such as shelter, food, clothes, and fuel. Genes involved in the assembly of the plant cell wall are often highly redundant. To identify these redundant genes, higher-order knock-out mutants need to be generated, which is conventionally done by genetic crossing. Compared with genetic crossing, CRISPR/Cas9 multi-gene targeting can greatly shorten the process of higher-order mutant generation and screening, which is especially useful to characterize cell wall related genes in plant species that require longer growth time. Moreover, CRISPR/Cas9 makes it possible to knock out genes when null T-DNA mutants are not available or are genetically linked. Because of these advantages, CRISPR/Cas9 is becoming an ideal and indispensable tool to perform functional studies in plant cell wall research. In this review, we provide perspectives on how to design CRISPR/Cas9 to achieve efficient gene editing and multi-gene targeting in plants. We also discuss the recent development of the virus-based CRISPR/Cas9 system and the application of CRISPR/Cas9 to knock in genes. Lastly, we summarized current progress on using CRISPR/Cas9 for the characterization of plant cell wall-related genes.


1994 ◽  
Vol 22 (2) ◽  
pp. 374-378 ◽  
Author(s):  
Peter Albersheim ◽  
Jinhua An ◽  
Glenn Freshour ◽  
Melvin S. Fuller ◽  
Rafael Guillen ◽  
...  

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