Expression of β-1,3-1,4-glucanase gene of Bacillus sp. SJ-10 in Lactococcus lactis using nisin-controlled expression [NICE] system

Abstract

β-1,3-1,4-glucanase from Bacillus sp. SJ-10 in Lactococcus lactis using a nisin-controlled expression NICE system was observed. Enzyme solubilization using E. coli has been studied, but the solubilization method of β-1,3-1,4-glucanase has a problem in that it takes a long time to purify through refolding process steps. In this study, β-1,3-1,4-glucanase gene isolated from Bacillus sp. SJ-10 was identified, and it was confirmed that the protein expressed by L. lactis NZ3900 was secreted out of the cell. Protein expression using the NICE system was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. The enzyme was purified from recombinant L. lactis and its activity was compared with E. coli. In addition, the degradation of barley (1,3)(1,4)-β-D-glucan was analyzed by Thin layer chromatography(TLC) using cell culture supernatant of L. lactis. To use β-oligosaccharides as a prebiotic, the food grade was constructed using the host and plasmid. Therefore, we confirmed the possibility of using it as a feed additive and food industry.