Purification and Some Properties of an Endo-β,1,6-glucanase fromNeurospora crassa

Purification and characterization of an endo-beta-1,6-glucanase from Trichoderma harzianum that is related to its mycoparasitism

“…A subtilisin-type serine proteinase induced by chitin has already been described in a mycoparasitic strain of Trichoderma harzianum (19). Also, ␤-1,6-glucanases (EC 3.2.1.75) have been shown to lyse yeast and fungal cell walls in filamentous fungi (41,51) and bacteria (39), but unfortunately there is almost no information about the purification and characterization of ␤-1,6-glucanases (20,39,41,51), and to the best of our knowledge, neither antibodies nor a sequence of a ␤-1,6-glucanase has ever been reported.…”

Cloning and Expression of an Endo-1,6-β-D-glucanase Gene (neg1) fromNeurospora crassa

“…3. The speciˆc activity of His-tagged recombinant Neg1 was 11.1 mmol W min W mg, that was nealy equal to that of native Neg1 (13.4 mmol W min W mg) 4) . When the Neg1 sequence is compared with those available in the databases, several glycosyl hydrolases show acceptable similarity scores (20-30z identity at the amino acid level).…”

“…We have previously puriˆed and characterized an endo-1,6-b-D-glucanase from Neurospora crassa, and the enzyme partially degraded N. crassa cell wall b-D-glucan, liberating small amounts of oligosaccharides 4) . In this paper, we report the cloning and expression of the endo-1,6-b-D-glucanase gene from N. crassa.…”

“…The endo-1,6-b-D-glucanase was puriˆed from the cultureˆltrate of Neurospora crassa IFO-6068 (grown for 48 h at 259 C in N-Vogel medium as described previously 4) ) by chromatographies on CM celluloˆne, Butyl toyopearl 650s, and a 2nd CMcelluloˆne. The enzyme was puriˆed 184-fold with a recovery of 24z.…”

“…Each fraction was incubated with 1,6-b-D-glucan for 30 min at 409 C in 0.1 M sodium acetate buŠer, pH 5.0, as described previously 4) . The reaction mixture was assayed for reducing sugar by the SomogyiNelson method 6) .…”