Banca de DEFESA: ANA AMÉLIA MAIA SILVA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : ANA AMÉLIA MAIA SILVA
DATE: 27/08/2021
TIME: 08:30
LOCAL: https://meet.google.com/iqq-gwfi-xxa
TITLE:

Expression of Cryptococcus flavus amylase in Pichia pastoris (Komagataella phaffii) cells for industrial applications

KEY WORDS:

Pichia pastoris; Komagataella phaffii; Cryptococcus flavus; Amylase; Heterologous Expression.

PAGES: 100
BIG AREA: Ciências Biológicas
AREA: Bioquímica
SUMMARY:

The expression of recombinant proteins through heterologous systems is an alternative for the production of recombinant enzymes. The available models have adequate machinery to increase the production levels of a certain protein, allow a faster production and with production levels higher than native systems, and provide biological safety to the product produced. This work refers to the construction of a strain of the yeast Pichia pastoris (Komagataella phaffii) for the overproduction of the enzyme amylase from Cryptococcus flavus, from methanol as a carbon source. The C. flavus α-amylase enzyme has interesting characteristics from a biotechnological point of view, such as thermal stability and maintaining its functional activity over a wide pH range. The rpAMY1 gene was synthesized in vitro with codons optimized for expression in K. phaffii in the pPICZα-B vector. From that, the genetic material was amplified and the yeast was transformed with the recombinant plasmid. Then, the starch hydrolysis potential of the pre-selected recombinant clones was assessed qualitatively and quantitatively. Preliminary results showed that the α-amylase gene was successfully expressed in K. phaffii cells and two recombinant clones are strong candidates for biotechnological applications of the expressed enzyme, as they showed higher enzyme activity values than those found in the literature, when expressed in their native host and Saccharomyces cerevisiae

BANKING MEMBERS:
Presidente - 1367304 - ALEXSANDRO SOBREIRA GALDINO
Interno - 2981680 - LEONARDO HENRIQUE FRANCA DE LIMA
Externa à Instituição - MARIA DE LOURDES BORBA MAGALHÃES - UDESC