Banca de DEFESA: Igor Patrick Vasconcelos Vieira

Uma banca de DEFESA de DOUTORADO foi cadastrada pelo programa.
STUDENT : Igor Patrick Vasconcelos Vieira
DATE: 16/05/2024
TIME: 10:00
LOCAL: Online
TITLE:

Development of new methanol-free regulation systems for protein production in Komagataella phaffii

KEY WORDS:

Komagataella phaffii; riboswitch; tetracycline; aptamer; translational control of gene expression. 

PAGES: 88
BIG AREA: Ciências Biológicas
AREA: Biologia Geral
SUMMARY:

 Komagataella phaffi (formerly known as Pichia pastoris) is considered one of the most important platforms available for large-scale production of proteins and metabolites today, due to its high productivity and easy control of fermentative parameters. Its most distinctive physiological feature is the ability to use methanol as a sole carbon source, achieved through the hyperexpression of the AOX1 gene encoding the peroxisomal alcohol oxidase enzyme. The PAOX1 promoter allows high expression with finely regulated control by methanol concentration, making it the most widely used recombinant expression system in this organism. However, the main limitation of this system in the context of industrial bioprocesses is the highly toxic and flammable nature of methanol. The development of a gene expression induction system based on a non-toxic inducer agent, preferably not consumed by the cell, is necessary to reduce production costs on a large scale. In this work, two expression control mechanisms were tested: at the transcriptional level we used the GAL system from S. cerevisiae to induce the production of green fluorescent protein (GFP) under the control of the PGAL1 promoter regulated by galactose and; at the translational level, expression control was achieved using a tetracycline-controlled riboswitch aiming to inhibit the production of β-galactosidase and Ku70p proteins. The results demonstrated success in using the riboswitch as a mechanism for controlling protein production in K. phaf with easy application and significant potential impact on the application of this system on a large scale. 

COMMITTEE MEMBERS:
Externa ao Programa - 2518326 - CINTIA MARQUES COELHO - nullPresidente - 1193623 - FERNANDO ARARIPE GONCALVES TORRES
Externo à Instituição - JOÃO RICARDO MOREIRA DE ALMEIDA - EMBRAPA
Externa ao Programa - 2221665 - LIDIA MARIA PEPE DE MORAES - nullExterna à Instituição - Milca Rachel da Costa Ribeiro Lins - UFABC