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Applied and Environmental Microbiology, September 2004, p. 5145-5152, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5145-5152.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Modulation of Aspergillus awamori Thaumatin Secretion by Modification of bipA Gene Expression

Marta Lombraña,1 Francisco J. Moralejo,1 Rosa Pinto,1 and Juan F. Martín1,2*

Instituto de Biotecnología de León (INBIOTEC),1 Área de Microbiología, Facultad de Ciencias Biológicas y Ambientales, Universidad de León, León, Spain2

Received 9 March 2004/ Accepted 6 May 2004

Two different strains, Aspergillus awamori TGDTh-4 and A. awamori TGP-3 overexpressing a synthetic gene encoding the plant sweet protein thaumatin, showed an unfolded protein response. To facilitate protein secretion, the chaperone BiPA gene was expressed in A. awamori under control of the strong constitutive promoter of the gpdA gene. A good correlation was observed between the level of the bipA transcript in different strains and the amount of thaumatin secreted. Thaumatin secretion was increased 2- to 2.5-fold in transformants overexpressing the bipA gene compared with the parental strain. Secretion of the homologous proteins {alpha}-amylase and glucoamylase was not affected by the bipA gene overexpression. The requirement for BiPA for secretion of thaumatin was confirmed by attenuation of the endogenous bipA gene expression with an antisense RNA cassette. The decrease in bipA expression reduced the amount of secreted thaumatin up to 80% without affecting the secretion of the homologous {alpha}-amylase and glucoamylase proteins. The BiPA protein is, therefore, very important for secretion of some heterologous proteins, such as thaumatin in A. awamori.

* Corresponding author. Mailing address: Área de Microbiología, Faculad de Ciencias Biológicas y Ambientales, Universidad de León, Campus de Vegazana s/n, 24071 León, Spain. Phone: 34 987 291 505. Fax: 34 987 291 506. E-mail: degjmm{at}unileon.es.

Applied and Environmental Microbiology, September 2004, p. 5145-5152, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5145-5152.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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