A new source of bacterial myrosinase isolated from endophytic Bacillus sp. NGB-B10, and its relevance in biological control activity.

Plant metabolism interacts strongly with the plant microbiome. Glucosinolates, secondary metabolites synthesized by Brassica plants, are hydrolyzed by myrosinase into bioactive compounds of great importance in human health and plant protection. Compared with myrosinase from plant sources, myrosinase enzymes of microbial origin have not been extensively investigated. Therefore, seven endophytic strains corresponding to Bacillus sp. were isolated from Eruca vesicaria ssp. sativa plants that could hydrolyse glucosinolates (sinigrin) in the culture medium and showed myrosinase activity (0.08-19.92 U mL-1). The bglA myrosinase-related gene encoding the 6-phospho-ß-glucosidase (GH 1) from Bacillus sp. NGB-B10, the most active myrosinase-producing bacterium, was successfully identified. Response surface methodology (RSM) was applied to statistically optimize culture conditions for myrosinase production from Bacillus sp. strain NGB-B10. The Plackett-Burman design indicated that nitrogen concentration, incubation period, and agitation speed were the significant parameters in myrosinase production. The application of the Box-Behnken design of RSM resulted in a 10.03-fold increase in enzyme activity as compared to the non-optimized culture conditions. The myrosinase was partially purified by 40% fractionation followed by SDS-PAGE analysis which yielded two subunits that had a molecular weight of 38.6 and 35.0 KDa. The purified enzyme was stable under a broad range of pH (5.5-10) and temperatures (10-65 °C). The hydrolysis products released by bacterial myrosinase from some glucosinolate extracts had higher and/or equivalent in vitro antagonistic activity against several phytopathogenic fungi compared to the nystatin (a broad-spectrum antifungal agent). This study provides original information about a new source of bacterial myrosinase and affords an optimized method to enhance myrosinase production.

Simultaneous biodegradation of λ-cyhalothrin pesticide and Vicia faba growth promotion under greenhouse conditions.

λ-cyhalothrin is a widely used synthetic pyrethroid insecticide and its persistence in plant, soil and water exerts a detrimental effect on humans as well as the environment. There are many studies regarding isolated bacteria capable of degrading λ-cyhalothrin in vitro. However, limited work has been done examining the microbial degradation of λ-cyhalothrin together with plant growth promotion under greenhouse conditions. In this study, 43 bacterial strains were isolated from heavily polluted soil with λ-cyhalothrin by the enrichment technique. The plant growth promotion characteristics of all isolates were evaluated. The results revealed that five isolates were potential in λ-cyhalothrin biodegradation at high concentration (1200 mg/L) within only 24 h together with their high plant growth promotion abilities. The morphological, biochemical and 16S rDNA sequence analyses identified the isolates as Bacillus subtilis strains. The GC/MS analysis revealed that the selected isolates reached high levels of degradation after only two days, the degradation percentage ranged from 95.72 to 99.52% after 48 h of incubation. Furthermore, the degradation pathway for complete detoxification and metabolism of λ-cyhalothrin was established. Moreover, greenhouse experiment was conducted, the results indicate that the application of seed coat significantly enhanced Vicia faba seedling growth and caused an increase from 38.4 to 40.2% percentage of fresh and dry weight, respectively compared to untreated control. All isolates were effective to remove the pesticide residues in Vicia faba seedlings and recorded the highest degradation percentage of 83.79 under greenhouse conditions. Therefore, it can be concluded that the Bacillus subtilis strains isolated in this study have a dual potential role in complete mineralization of λ-cyhalothrin residues in vivo as well as effective biofertilization for future use in sustainable agriculture.