Purification and functional characterization of copper mine microbial oxidoreductases enzymes

• Grant number: 11/10378-5
• Support type: Scholarships in Brazil - Doctorate
• Effective date (Start): June 01, 2011
• Effective date (End): February 28, 2015
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Organic Chemistry
• Cooperation agreement: Vale-FAPEMIG-FAPESPA
• Principal Investigator: Anita Jocelyne Marsaioli
• Grantee: Bruna Zucoloto da Costa
• Home Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Company: Universidade Estadual de Campinas (UNICAMP). Instituto de Química (IQ)

Abstract

Biocatalytic processes have been addressed in this thesis from microorganism screening for the selection of suitable biocatalysts to the use of isolated enzymes in biotechnological reactions. The first chapter presents the isolation, identification and enzymatic screening of heterotrophic bacteria isolated from copper mine drainages. From these samples, 189 bacteria were isolated, that showed diverse catalytic activities. The isolated bacteria were identified by MALDI-TOF and 16S rRNA gene sequencing and several genera were found: Bacillus, Acinetobacter, Pseudomonas, Delftia, Stenotrophomonas, Hydrogenophaga, Rhodococcus, among others. The second chapter presents the catalytic potential of a new recombinant epoxide hydrolase from Aspergillus brasiliensis CCT1435 (AbEH), expressed in E. coli. This EH is active in a wide pH and temperature range, with a great performance at pH 6.0 and 30 °C. This work also presents the AbEH biocatalytic application for the styrene oxide hydrolysis in aqueous and biphasic medium. Finally, the third chapter presents preliminary results involving a chemo-enzymatic method for the production of pyrrolidine alkaloids from the corresponding 1,4-diketones. This approach is promising since several 1,4-diketones were converted to their respective cyclic imines, and the subsequent reduction with NaBH3CN promoted the formation of pyrrolidines. Therefore, this thesis presents a broad and diverse study concerning biocatalytic processes applied in a number of interesting biotechnological reactions. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SIGRIST, RENATA; DA COSTA, BRUNA ZUCOLOTO; MARSAIOLI, ANITA JOCELYNE; DE OLIVEIRA, LUCIANA GONZAGA. Nature-inspired enzymatic cascades to build valuable compounds. BIOTECHNOLOGY ADVANCES, v. 33, n. 5, SI, p. 394-411, SEP-OCT 2015. Web of Science Citations: 18.

BELOTI, LILIAN L.; COSTA, BRUNA Z.; TOLEDO, MARCELO A. S.; SANTOS, CLELTON A.; CRUCELLO, ALINE; FAVARO, MARIANNA T. P.; SANTIAGO, ANDRE S.; MENDES, JULIANO S.; MARSAIOLI, ANITA J.; SOUZA, ANETE P. A novel and enantioselective epoxide hydrolase from Aspergillus brasiliensis CCT 1435: Purification and characterization. Protein Expression and Purification, v. 91, n. 2, p. 175-183, OCT 2013. Web of Science Citations: 10.

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

COSTA, Bruna Zucoloto da. Biocatalytic processes applying epoxide hydrolases, oxidoreductases and transaminases. 2015. Doctoral Thesis - Universidade Estadual de Campinas. Instituto de Química.