000			02114nam a2200205Ia 4500
003			MX-MdCICY
005			20250625140615.0
040			_cCICY
090			_aB-9597
245	1	0	_aA new method for screening and isolation of hypersecretion mutants in Aspergillus niger
490	0		_vAppl Microbiol Biotechnol, 69(6), p.711-717, 2006
520	3		_aAlthough filamentous fungi have a unique property of secreting a large amount of homologous extracellular proteins, the use of filamentous fungi as hosts for the production of heterologous proteins is limited because of the low production levels that are generally reached. Here, we report a general screening method for the isolation of mutants with increased protein production levels. The screening method makes use of an Aspergillus niger strain that lacks the two major amylolytic enzymes, glucoamylase (GlaA)and acid amylase (AamA). The double-mutant strain grows poorly on starch and its growth is restored after reintroducing the catalytic part of the glucoamylase gene (GlaA512). We show that the fusion of a heterologous protein, a laccase from Pleurotus ostreatus (Pox2), to the catalytic part of glucoamylase (GlaA512-Pox2)severely hampers efficient production of the glucoamylase protein, resulting in a slow-growth phenotype on starch. Laccasehypersecreting mutants were obtained by isolating mutants that displayed improved growth on starch plates. The mutant with the highest growth rate on starch displayed the highest laccase activity, indicating that increased glucoamylase protein levels are correlated with higher laccase production levels. In principle, our method can be applied to any low-produced heterologous protein that is secreted as a fusion with the glucoamylase protein.
700	1	2	_aWeenink, X.O.
700	1	2	_aPunt, P.J.
700	1	2	_aVan Den Hondel, C.A.M.J.J.
700	1	2	_aRam, A.F.J.
856	4	0	_uhttps://drive.google.com/file/d/11-5AO2mWfhKjuaZPDsVmxqkqLmHsWSY4/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
942			_2Loc _cREF1
008			250602s9999 xx |||||s2 |||| ||und|d
999			_c43844 _d43844