Expression of β (1-3)(1-4) glucanase gene in Lactococcus lactis to produce an animal probiotic feed

Probiotics are bacteria with beneficial effects to humans and animals. Gram-positive Lactococcus lactis can be genetically engineered to efficiently produce a large variety of proteins. Glucanase enzymes, are capable to hydrolyze β-glucans (a non-starch polysaccharide). β(1,3)-(1,4) glucan (Lechinan) is a characteristic hemicellulose in primary cell walls of grasses such as barley, rye, sorghum, rice and wheat. The purpose of this study is to express the lechinase enzyme in Lactococcus lactis to achieve a feed supplement for hydrolyzation of barley β-glucan to replace corn by barley in poultry diets. licBM2 gene encoding lechinas enzyme with a secretion signal peptide was cloned in pNZ8149 plasmid. Then recombinant plasmids were transferred to Lactococcus lactis using electroporation method and the transformed colonies were selected via their ability to grow on lactose containing medium. The expression of transgene was induced using of Nisin in the medium. Our results showed that by increasing of the induction time, the expression of lechinase gene and consequently the production of enzyme in the medium were increased.