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Study of Protoplast Transformation Technique in Bacillus megaterium for Recombinant Penicillin G Acylase Production.

Grant number: 11/15864-5
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): October 01, 2011
Effective date (End): September 30, 2012
Field of knowledge:Engineering - Chemical Engineering
Principal researcher:Rosineide Gomes da Silva Cruz
Grantee:Fabio dos Santos Machado Zanucoli
Home Institution: Centro de Ciências Exatas e de Tecnologia (CCET). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil

Abstract

Penicillin G acylase (PGA) is a very important catalyst for the pharmaceutical industry. Its main application is in the hydrolysis of penicillin G, generating 6-aminopenicillanic acid, a precursor of semi-synthetic antibiotics. The focus of this project is the application of molecular biology techniques to the penicillin G acylase production process by Bacillus megaterium. The project aims at study of protoplast transformation technique in B. megaterium, which unlike E. coli is Gram-positive microorganism. The microorganism transformation is a critical step in recombinant DNA technology. Depending on the microorganism, one technique can be more appropriate than another. In this project, the B. megaterium cells growth conditions and their preparations to protoplasts transformation will be investigated for PGA expression.The proponent is member of a research group and with access to a well-equipped molecular biology laboratory. One of the main research in this laboratory includes recombinant PGA production. Nowadays, the group has one post-doctoral expert in this area, two PhD students in progress and contributes with different groups in Brazil and in the world. This undergraduate research project is part of researches involving the recombinant PGA production that has been done in Chemical Engineering Department at UFSCar. In the FAPESP project recently approved ("Study of transformation techniques of the different hosts for the recombinant penicillin G acylase production", # 2010/15916-2) two undergraduate research themes, one of these is been requested in this proposal, was foreseen. In addition to the progress in the technology focusing an enzyme of great interest for the pharmaceutical industry, this project will enable the undergraduate research student to work in biochemistry and molecular biology in order to enrich and complement their academic formation.

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