Record E, Asther M, Sigoillot C, Pagès S, Punt P J, Delattre M, Haon M, van den Hondel C A M J J, Sigoillot J-C, Lesage-Meessen L, Asther Marcel
UMR 1163 INRA/Université de Provence de Biotechnologie des Champignons Filamenteux, IFR-BAIM, Universités de Provence et de la Méditerranée, ESIL, 163 avenue de Luminy, Case Postale 925, 13288, Marseille Cedex 09, France.
Appl Microbiol Biotechnol. 2003 Sep;62(4):349-55. doi: 10.1007/s00253-003-1325-4. Epub 2003 May 13.
A well-known industrial fungus for enzyme production, Aspergillus niger, was selected to produce the feruloyl esterase FAEA by homologous overexpression for pulp bleaching application. The gpd gene promoter was used to drive FAEA expression. Changing the nature and concentration of the carbon source nature (maltose to glucose; from 2.5 to 60 g l(-1)), improved FAEA activity 24.5-fold and a yield of 1 g l(-1) of the corresponding protein in the culture medium was achieved. The secreted FAEA was purified 3.5-fold to homogeneity in a two-step purification procedure with a recovery of 69%. The overproduced protein was characterised and presented properties in good agreement with those of native FAEA. The recombinant FAEA was tested for wheat straw pulp bleaching, with or without a laccase mediator system and xylanase. Best results were obtained using a bi-sequential process with a sequence including xylanase, FAEA and laccase, and yielded very efficient delignification--close to 75%--and a kappa number of 3.9. This is the first report on the potential application of recombinant FAEA in the pulp and paper sector.
黑曲霉是一种著名的用于酶生产的工业真菌，通过同源过表达来生产阿魏酸酯酶FAEA，用于纸浆漂白。gpd基因启动子用于驱动FAEA的表达。改变碳源的性质和浓度（从麦芽糖改为葡萄糖；从2.5 g/L至60 g/L），FAEA活性提高了24.5倍，并在培养基中实现了1 g/L相应蛋白质的产量。分泌的FAEA通过两步纯化程序纯化至同质，纯化倍数为3.5倍，回收率为69%。对过量生产的蛋白质进行了表征，其特性与天然FAEA的特性高度一致。对重组FAEA进行了麦草浆漂白测试，分别测试了有无漆酶介导系统和木聚糖酶的情况。采用包括木聚糖酶、FAEA和漆酶的两步连续工艺获得了最佳结果，实现了非常高效的脱木质素——接近75%——以及卡伯值为3.9。这是关于重组FAEA在制浆造纸行业潜在应用的首次报道。
