Cellulases and related enzymes in biotechnology

Broad spectrum biological control of diseases caused by soilborne plant pathogens such as Pythium, Phytophthora, and Rhizoctonia solani requires the introduction into or presence of edaphic sources of organic nutrients in soil for sustenance of biocontrol agents. The decomposition level of organic matter critically affects the composition of bacterial taxa as well as the populations and activities of biocontrol agents. Competition, antibiosis, parasitism, and systemic induced resistance are all affected. Highly stabilized sources of Sphagnum peat consistently fail to support sustained biological control, even when inoculated with biocontrol agents. Composts, on the other hand, can serve as an ideal food base for biocontrol agents and offer an opportunity to introduce and establish specific biocontrol agents into soils, which in turn leads to sustained biological control based on the activities of microbial communities.

BIOCONTROL WITHIN THE CONTEXT OF SOIL MICROBIAL COMMUNITIES: A Substrate-Dependent Phenomenon.

The contribution of cell wall proteins to the organization of the yeast cell wall

Ligands for the β-glucan receptor, dectin-1, assigned using "designer" microarrays of oligosaccharide probes (neoglycolipids) generated from glucan polysaccharides

State of the art and future prospects of the biological control of postharvest fruit diseases.

The mycoparasitic fungus Trichoderma harzianum CECT 2413 produces at least three extracellular beta-1,3-glucanases. The most basic of these extracellular enzymes, named BGN13.1, was expressed when either fungal cell wall polymers or autoclaved mycelia from different fungi were used as the carbon source. BGN13.1 was purified to electrophoretic homogeneity and was biochemically characterized. The enzyme was specific for beta-1,3 linkages and has an endolytic mode of action. A synthetic oligonucleotide primer based on the sequence of an internal peptide was designed to clone the cDNA corresponding to BGN13.1. The deduced amino acid sequence predicted a molecular mass of 78 kDa for the mature protein. Analysis of the amino acid sequence indicates that the enzyme contains three regions, one N-terminal leader sequence; another, nondefined sequence; and one cysteine-rich C-terminal sequence. Sequence comparison shows that this beta-1,3-glucanase, first described for filamentous fungi, belongs to a family different from that of its previously described bacterial, yeast, and plant counterparts. Enzymatic-activity, protein, and mRNA data indicated that bgn13.1 is repressed by glucose and induced by either fungal cell wall polymers or autoclaved yeast cells and mycelia. Finally, experimental evidence showed that the enzyme hydrolyzes yeast and fungal cell walls.

A novel endo-beta -1,3-glucanase, bgn13.1, involved in the mycoparasitism of trichoderma harzianum

The enzymes from Trichoderma species that degrade fungal cell walls have been suggested to play an important role in mycoparasitic action against fungal plant pathogens. The mycoparasite Trichoderma harzianum produces at least two extracellular beta-1,6-glucanases, among other hydrolases, when it is grown on chitin as the sole carbon source. One of these extracellular enzymes was purified to homogeneity after adsorption to its substrate, pustulan, chromatofocusing, and, finally, gel filtration. The apparent molecular mass was 43,000, and the isoelectric point was 5.8. The first 15 amino acids from the N terminus of the purified protein have been sequenced. The enzyme was specific for beta-1,6 linkages and showed an endolytic mode of action on pustulan. Further characterization indicated that the enzyme by itself releases soluble sugars and produces hydrolytic halli on yeast cell walls. When combined with other T. harzianum cell wall-degrading enzymes such as beta-1,3-glucanases and chitinases, it hydrolyzes filamentous fungal cell walls. The enzyme acts cooperatively with the latter enzymes, inhibiting the growth of the fungi tested. Antibodies against the purified protein also indicated that the two identified beta-1,6-glucanases are not immunologically related and are probably encoded by two different genes.

T Benítez

Papers

A novel endo-beta -1,3-glucanase, bgn13.1, involved in the mycoparasitism of trichoderma harzianum

Purification and characterization of an endo-beta-1,6-glucanase from Trichoderma harzianum that is related to its mycoparasitism.