Exploring of the inulin synthesis gene of endophytic bacteria by the new degenerated primers

DOI: 10.5176/2251-2489_BioTech16.13

Authors: Sumolnat Khamwan and Wiyada Mongkolthanaruk

Abstract: This study aims to isolate and identify inulin synthesis of endophytic bacteria from Jerusalem artichoke. Forty-six isolates of endophytic bacteria were studied for characterization of inulin synthesis gene by polymerase chain reaction technique with the new primers designed on conserved amino acids of bacteria produced inulosucrase gene. The degenerated primers were Ins_F (5’ CAAGAATGGTCDGGRTCWGCK 3’) and Ins_R (5’ NGGRACWGCRTAATAWGAATAWGT 3’). The endophytic bacteria isolate 3.5, identified as Bacillus megaterium, contained the gene fragment of 800 base pair which was cloned and sequenced to verify inulin synthesis gene. The results showed that the gene fragment encoded 278 amino acids were levansucrase with 96{6e6090cdd558c53a8bc18225ef4499fead9160abd3419ad4f137e902b483c465} similarity. The levansucrase is inulin synthesis enzyme in a glycoside hydrolase family 68 (GH68). The new primers were suitable to explore inulin synthesis of bacteria.

Keywords: Fructan; inulin; inulosucrase; levansucrase