Conserved regions and molecular cloning of Acid and Alkaline phosphatases in Streptomyces sp. MMA-NRC

Abstract Phosphorus is one of the most important nutrients for the growth and development of plants. Although chemical fertilizers supply phosphorus, much of it remains in insoluble forms inaccessible to plants. Numerous applications exist for phosphate-solubilizing bacteria as biological fertilizers in agriculture. By employing sustainable agricultural practices, the use of biofertilizers rather than artificial fertilizers can reduce environmental contamination and increase crop productivity. Here, we explored conserved regions and the gene encoding acid phosphatase ( ACPase ) and alkaline phosphatase ( ALPase ) sequences from Streptomyces sp. MMA-NRC , the specimen was separated; genes were sequenced, annotated, and officially recorded in the NCBI GenBank database with unique identification numbers PV646716 and PV646717. Ramachandran’s plot was used to validate the 3D structure of the simulated ACPase and ALPase proteins. The results showed that a significant portion of the amino acid residues were located in the most favored region for the strain Streptomyces NM-NRC , which revealed that 488 and 560 residues, accounting for 94.37 and 93.08% overall quality factor for ACPase and ALPase; respectively. strain 4 F and strain Streptomyces sp. MMA-NRC. Docking studies revealed optimal binding affinities of the ACPase and ALPase proteins with rock phosphate, with high affinity scores for the strain Streptomyces sp. MMA-NRC . Which recorded an affinity score of -110.86 kcal/mol, a confidence score of 0.3137, a ligand RSMD (Å) of 69.31, for wild type strain Streptomyces sp. MMA-NRC ACPase . In addition, recorded affinity score of − 108.55 kcal/mol, confidence score of 0.3039, ligand RSMD (Å) of 33.68, for the wild type strain Streptomyces sp. MMA-NRC ALPase strain. Molecular cloning and expression of the Streptomyces sp. MMA-NRC ACPase and ALPase genes in E. coli DH5α resulted in a recombinant E. coli DH5α pGEM- T-ACPase and E. coli DH5α pGEM- T-ALPase significantly higher available phosphore using ascorbic acid colorimetric method of 52.64 and 57.22 mg/L − 1 ; respectively after 7 days of incubation, compared to the wild type Streptomyces sp. MMA-NRC , which exhibited a value 35.44 mg/L − 1 after 7 days of incubation. These results suggest that the use of Streptomyces sp. MMA-NRC for phosphatase production seems to be very promising since it has a diverse array of agricultural uses, including plant biofertilizers.