RT info:eu-repo/semantics/article
T1 Thaumatin Production in Aspergillus awamori by Use of Expression Cassettes with Strong Fungal Promoters and High Gene Dosage
A1 Moralejo, Francisco J.
A1 Cardoza, Rosa E.
A1 Gutiérrez Martín, Santiago
A1 Martín Martín, Juan Francisco
A2 Microbiologia
K1 Biología
K1 Heterologous expression
K1 Filamentous fungi
K1 Thaumatin
K1 2414 Microbiología
K1 2414.01 Antibióticos
K1 2414.06 Hongos
K1 2414.07 Metabolismo Microbiano
K1 2415.01 Biología Molecular de Microorganismos
AB [EN] Four expression cassettes containing strong fungal promoters, a signal sequence for protein translocation, a KEX protease cleavage site, and a synthetic gene (tha) encoding the sweet protein thaumatin II were used to overexpress this protein in Aspergillus awamori lpr66, a PepA protease-deficient strain. The best expression results were obtained with the gdhA promoter ofA. awamori or with the gpdA promoter ofAspergillus nidulans. There was good correlation oftha gene dosage, transcript levels, and thaumatin secretion. The thaumatin gene was expressed as a transcript of the expected size in each construction (1.9 or 1.4 kb), and the transcript levels and thaumatin production rate decayed at the end of the growth phase, except in the double transformant TB2b1-44-GD5, in which secretion of thaumatin continued until 96 h. The recombinant thaumatin secreted by a high-production transformant was purified to homogeneity, giving one major component and two minor components. In all cases, cleavage of the fused protein occurred at the KEX recognition sequence. This work provides new expression systems inA. awamori that result in very high levels of thaumatin production.
PB American Society for Microbiology ASM Journals
SN 0099-2240
LK https://hdl.handle.net/10612/17796
UL https://hdl.handle.net/10612/17796
NO Moralejo, F., Cardoza, R. E., Gutiérrez, S. y Martı́n, J. F. (1999). Thaumatin production in Aspergillus awamori by use of expression cassettes with strong fungal promoters and high gene dosage. Applied and Environmental Microbiology, 65(3), 1168-1174. https://doi.org/10.1128/aem.65.3.1168-1174.1999
NO https://journals.asm.org/doi/10.1128/aem.65.3.1168-1174.1999
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 03-jun-2024