Recombinant Hypocrea rufa Endo-beta-1,6-galactanase (6GAL)

The gene fragment corresponding to the 21-479aa of the Hypocrea rufa 6GAL protein was synthesized, with appropriate restriction sites suitable for in-frame cloning into an expression vector, with N-terminal 10xHis tag. The E.coli was transformed with the expression vector, and the clone was expressed upon certain induction. After the induced cell centrifugation, the recombinant protein was purified from the cell extract and presented as N-terminal 10xHis-tagged fusion. Its purity is greater than 85% assayed by SDS-PAGE. The 6GAL protein ran to a band of about 55 kDa molecular weight on the gel.

To date, the research in terms of 6GAL is quite less. There are two references about 6GAL on Uniport: one is "Molecular cloning and expression in Escherichia coli of a Trichoderma viride endo-beta-(1-->6)-galactanase gene" The other is "Purification and characterization of an endo-beta-(1-->6)-galactanase from Trichoderma viride". Besides, the research of 6GAL from the NCBI data also provde some information about 6GAL. In certain article, the putative endo-β-1,6-galactanase gene from Streptomyces avermitilis was cloned and expressed in Escherichia coli. The enzymatic properties of the recombinant enzyme were characterized. The enzyme produced galactose and a range of β-1,6-linked galacto-oligosaccharides, predominantly β-1,6-galactobiose, from β-1,6-galactan chains. In a early resport, it has been found that treatment of larch wood arabinogalactan with the recombinant enzyme indicated that FoGal1 is a β-1,6-galactanase that preferentially debranches β-1,6-galactobiose from the substrate.