Search Results (5)

Phytases of the PhyD class according to their pH optimum (7.0–7.8) and high thermal stability can claim to be used in the production of feed supplements. However, today they have no practical application in feed production because there are no suitable producers sufficient for its biotechnological production compared to the PhyA and PhyC class ones. Moreover, in most cases, the technologies with the enzymes produced in secretory form are preferable for the production of phytases, though upon microencapsulation in yeast-producing cells, the phytase thermal stability increases significantly compared to the extracellular form, which improves its compatibility with spray drying technology. In this study, we assayed the intracellular heterologous expression of PhyD phytase from Bacillus species in the Yarrowia lipolytica yeast cells. While the technology has been successfully used to synthesize PhyC phytase from Obesumbacterium proteus, PhyD phytase tends to aggregate upon intracellular accumulation. Furthermore, we evaluated the prospects for the production of encapsulated phytase of the PhyD class of high enzymatic activity when it accumulates in the cell cytoplasm of the Y. lipolytica extremophile yeast, a highly effective platform for the production of recombinant proteins. Full article

The widespread use of non-renewable phosphate fertilizers in agriculture poses a significant pollution threat to soil, necessitating the exploration of sustainable alternatives for phosphate fertility. Releasing phytate phosphorus through microbial phytases presents an eco-friendly solution for sustainable phosphate fertility in agriculture. This study directly inoculated dual-domain β-propeller alkaline phytase (phyHT) derived from Bacillus sp. HJB17 into the soil. The study analyzed the impact of inoculated phyHT on the physicochemical properties of the soil, assessed the variations in enzyme activity of phyHT within the soil, and examined the effects of the treated soil on wheat growth. Additionally, the study explored the enhancement of the available phosphorus in the soil through the inoculation of phyHT in both crop residues and organic fertilizer. PhyHT exhibited the highest catalytic activity at 37 °C and pH 8.0. After soil adsorption, phyHT maintained stable enzymatic activity. PhyHT markedly boosted the available phosphorus in the soil while reducing the soil phytate content by about 20%, increasing the phosphorus levels and enhancing soil fertility. PhyHT effectively degraded phytates in an organic fertilizer and crop residues, increasing the available phosphorus. PhyHT supplementation enhanced growth, biomass, and phosphorus content in both the shoot and root weights of Triticum aestivum. This study establishes phyHT as a viable and eco-friendly method to enhance phosphorus fertility in soil. The direct application of microbial phytases can serve as a sustainable source of phosphate fertility in soil. Full article

The growth of the aquafeed sector is highly dependent upon the availability of fish feed with a balanced nutritional composition. The use of prebiotics and probiotics can be an effective solution to increase the bioavailability of feed components. In this study, we evaluated the effect of dietary supplementation with β-propeller phytase (0, 600, 1200, 1800 and 2400 U/kg) from Bacillus and mannan oligosaccharide (MOS) (0, 2, 4 and 8 g/kg) on growth performance and nutrient digestibility of Nile tilapia over 45 days. The findings showed that adding phytase significantly (p < 0.05) increases the growth performance and nutrient digestibility; the 1200 and 1800 U/kg PHY levels showed the maximum weight gain (WG), specific growth rate (SGR) and protein efficiency ratio (PER) and best feed conversion ratio (FCR). Furthermore, phytase increases carcass mineral composition (phosphate and calcium). At the end of the experiment, there were no significant differences among all feeding groups in survival rates (above 90%). Regarding MOS inclusion, insignificant differences were seen in WG, SGR and SR. However, significant effects of MOS were observed on FCR, feed intake (FI) and PER when supplemented at 4 and 8 g/kg of feed. Taken together, our results suggest that supplementation of Nile tilapia feed with adequate amounts of β-propeller phytase from Bacillus and MOS increases growth performance and nutrient digestibility. Full article

Aquaponics are efficient systems that associate aquatic organisms’ production and plants by recirculating water and nutrients between aquaculture and hydroponic tanks. In this study, we characterised the bacterial communities in the freshwater aquaponics system that can mineralise polysaccharides and phytate by producing carbohydrate-degrading enzymes and phytases, by 16S rRNA gene sequencing and in vitro culture techniques. Around 20% of the operational taxonomic units (zOTUs) identified were previously reported to carry fibre-degrading enzyme putative genes, namely β-glucanase (1%), xylanase (5%), or cellulases (17%). Ten % of the zOTUs were previously reported to carry putative genes of phytases with different catalytic mechanisms, namely β-propeller (6%), histidine acid phytases (3%), and protein tyrosine phytase (<1%). Thirty-eight morphologically different bacteria were isolated from biofilms accumulated in fish and plant compartments, and identified to belong to the Bacilli class. Among these, 7 could produce xylanase, 8 produced β-glucanase, 14 produced cellulase, and 11 isolates could secrete amylases. In addition, Staphylococcus sp. and Rossellomorea sp. could produce consistent extracellular phytate-degrading activity. The PCR amplification of β-propeller genes both in environmental samples and in the isolates obtained showed that this is the most ecologically relevant phytase type in the aquaponics systems used. In summary, the aquaponics system is abundant with bacteria carrying enzymes responsible for plant-nutrient mineralisation. Full article

Alkaline phytases from uncultured microorganisms, which hydrolyze phytate to less phosphorylated myo-inositols and inorganic phosphate, have great potential as additives in agricultural industry. The development of metagenomics has stemmed from the ineluctable evidence that as-yet-uncultured microorganisms represent the vast majority of organisms in most environments on earth. In this study, a gene encoding a phytase was cloned from red rice crop residues and castor bean cake using a metagenomics strategy. The amino acid identity between this gene and its closest published counterparts is lower than 60%. The phytase was named PhyRC001 and was biochemically characterized. This recombinant protein showed activity on sodium phytate, indicating that PhyRC001 is a hydrolase enzyme. The enzymatic activity was optimal at a pH of 7.0 and at a temperature of 35 °C. β-propeller phytases possess great potential as feed additives because they are the only type of phytase with high activity at neutral pH. Therefore, to explore and exploit the underlying mechanism for β-propeller phytase functions could be of great benefit to biotechnology. Full article