BGN16.3, a novel acidic β-1,6-glucanase from mycoparasitic fungus Trichoderma harzianum CECT 2413

Abstract

A new component of the β-1,6-glucanase (EC 3.2.1.75) multienzymatic complex secreted by Trichoderma harzianum has been identified and fully characterized. The protein, namely BGN16.3, is the third isozyme displaying endo-β-1,6-glucanase activity described up to now in T. harzianum CECT 2413. BGN16.3 is an acidic β-1,6-glucanase that is specifically induced by the presence of fungal cell walls in T. harzianum growth media. The protein was purified to electrophoretical homogenity using its affinity to β-1,6-glucan as first purification step, followed by chomatofocusing and gel filtration. BGN16.3 has a molecular mass of 46 kDa in SDS/PAGE and a pi of 4.5. The enzyme only showed activity against substrates with β-1,6-glycosidic linkages, and it has an endohydrolytic mode of action as shown by HPLC analysis of the products of pustulan hydrolysis. The expression profile analysis of BGN16.3 showed a carbon source control of the accumulation of the enzyme, which is fast and strongly induced by fungal cell walls, a condition often regarded as mycoparasitic simulation. The likely involvement β-1,6-glucanases in this process is discussed. © 2005 FEBS.