Cloning of levansucrase from Leuconostoc mesenteroides and its expression in Pichia pastoris

A gene (m1ft) encoding levansucrase from Leuconostoc mesenteroides has been previously cloned in Escherichia coli. Presently, m1ft was cloned and secretively expressed in Pichia pastoris. Methanol induction of recombinant M1FT resulted in the production of active levansucrase (PM1FT). PM1FT-5 was expressed as an active form, but the protein accumulated mainly inside the cells, representing about 5% of total cell proteins. M1FT was secreted into the culture supernatant with a maximum yield of 14,400 U/L using fed-batch fermentations. P. pastoris-derived M1FT displayed catalytic activities comparable to those of the E. coli-derived M1FT. PM1FT was glycosylated at its 2 potential N-glycosylation sites.