Isolation of β-1,3-Glucanase-Producing Microorganisms from Poria cocos Cultivation Soil via Molecular Biology

Abstract

β-1,3-Glucanase is considered as a useful enzymatic tool for β-1,3-glucan degradation to produce (1→3)-linked β-glucan oligosaccharides with pharmacological activity properties. To validly isolate β-1,3-glucanase-producing microorganisms, the soil of Wolfiporia extensa, considered an environment rich in β-1,3-glucan-degrading microorganisms, was subjected to high throughput sequencing. The results demonstrated that the genera Streptomyces (1.90%) and Arthrobacter (0.78%) belonging to the order Actinomycetales (8.64%) in the phylum Actinobacteria (18.64%) were observed in soil for P. cocos cultivation (FTL1). Actinomycetes were considered as the candidates for isolation of glucan-degrading microorganisms. Out of 58 isolates, only 11 exhibited β-1,3-glucan-degrading activity. The isolate SYBCQL belonging to the genus Kitasatospora with β-1,3-glucan-degrading activity was found and reported for the first time and the isolate SYBC17 displayed the highest yield (1.02 U/mg) among the isolates. To check the β-1,3-glucanase contribution to β-1,3-glucan-degrading activity, two genes, 17-W and 17-Q, encoding β-1,3-glucanase in SYBC17 and one gene QLK1 in SYBCQL were cloned and expressed for verification at the molecular level. Our findings collectively showed that the isolates able to secrete β-1,3-glucanase could be obtained with the assistance of high-throughput sequencing and genes expression analysis. These methods provided technical support for isolating β-1,3-glucanase-producing microorganisms.

Calcium regulates the mycophagous ability of Burkholderia gladioli strain NGJ1 in a type III secretion system-dependent manner

“…The phylogenetic analysis revealed that the endo-β-1, 3glucanase of NGJ1 has similarity with that of various Burkholderia sp. as well as some of the previously reported glucanases [50] of other bacteria (Additional file: Fig. S9).…”

“…Neighbor-Joining clustering and BioNJ algorithms were applied to a matrix of pairwise distances estimated using the maximum composite Likelihood (MCL) approach to construct an initial tree for the heuristic search. Similarly, phylogenetic tree reflecting the similarity of endo-β-1, 3-glucanase of NGJ1 with that of previously reported glucanases of other bacteria [50] was constructed.…”

A Comparative Analysis of Ash Leaf-Colonizing Bacterial Communities Identifies Putative Antagonists of Hymenoscyphus fraxineus

“…Bacteria directly attack fungal pathogens by secreting antibiotic compounds, including heterocyclic phenazines (Mavrodi et al, 2006) and cyclic lipopeptides (Ongena et al, 2007) or hydrogen cyanide as a volatile compound that acts directly on cells by blocking cytochrome c oxidase. The production of extracellular hydrolytic enzymes has an important role in degrading the cell wall polymers of fungal plant pathogens (Wu et al, 2018). However, the fungal pathogen is also suppressed through competitions for resources such as nutrients or space (e.g., by siderophores), niche colonization, blockage of the potential entry points of the pathogen, or prevention of the germination of its propagules, as well as indirectly through the induction of systemic resistance (Terhonen et al, 2018;Schlechter et al, 2019).…”

The Cysteine-Rich Repeat Protein TaCRR1 Participates in Defense against Both Rhizoctonia cerealis and Bipolaris sorokiniana in Wheat

“…Chitinases also have the secondary effect of releasing chitin, which functions as a PAMP [ 9 ]. β-1,3-Glucanase is able to hydrolyze β-1,3-glucan linker, a major structural cell wall component of the fungal cell wall, and to suppress fungal growth; thus, the enzyme acts as a defense molecule against fungal pathogens in plants [ 10 ]. Defensin can permeabilize fungal plasma membranes, induce Ca 2+ influx, and disrupt a Ca 2+ gradient essential for polar growth of hyphal tips [ 11 ].…”