Biological safety assessment of mutant variant of Allium sativum leaf agglutinin (mASAL), a novel antifungal protein for future transgenic application.
Genetic engineering has established itself to be an important tool for crop improvement. Despite the success, there is always a risk of food allergy induced by alien gene products. The present study assessed the biosafety of mutant Allium sativum leaf agglutinin (mASAL), a potent antifungal protein generated by site directed mutagenesis of Allium sativum leaf agglutinin (ASAL). mASAL was cloned in pET28a+ and expressed in E. coli, and the safety assessment was carried out according to the FAO/WHO guideline (2001). Bioinformatics analysis, pepsin digestion, and thermal stability assay showed the protein to be nonallergenic. Targeted sera screening revealed no significant IgE affinity of mASAL. Furthermore, mASAL sensitized Balb/c mice showed normal histopathology of lung and gut tissue. All results indicated the least possibility of mASAL being an allergen. Thus, mASAL appears to be a promising antifungal candidate protein suitable for agronomical biotechnology.
Division of Plant Biology, Bose Institute, Centenary Campus , P1/12, CIT Scheme, VIIM, Kankurgachi, Kolkata 700054, West Bengal, India.
Prithwi
P
Ghosh
Amit
A
Roy
Joydeep
J
Chakraborty
Sampa
S
Das
D000373:Agglutinins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D000935:Antifungal Agents / Q000737:chemistry / Q000276:immunology*; D005260:Female; D005737:Garlic / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D018515:Plant Leaves / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D055550:Protein Stability
D000373:Agglutinins; D000935:Antifungal Agents