Figure 2. Purification of lacasses produced by submerged culture of T . pubescens using coffee husk as substrate.

All samples were analyzed by SDS-PAGE under non-reducing conditions and stained with ABTS. No heat denaturation of the samples was conducted. (a) The crude extract (1) was centrifuged and filtered through a Whatman No. 1 filter (2), 0.45 µM membrane (3) 0.22 µm membrane (4), and ultrafiltrated through a 10 kDa cut-off membrane, permeate (5) and retentate (6). (b) UF retentate was purified by anionic exchange chromatography. Lac1 represent the unbound fraction, while Lac2 represent the eluted fraction. The arrow shows the point when protein elution was started. (c) Purified fractions (1) crude extract, (2) Lac1 and (3) Lac2. MW: Molecular Weight. Similar results were observed for crude extracts produced by using soybean pod husk and cedar sawdust.