Milena Andreotti Minetto

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Áreas de atuação

Participação em eventos

Produções bibliográficas

• FREITAS, M. A. ; FERREIRA, C. V. ; ABRANTES, J. L. F. . Butyrate Efficiently Diminishes Resistant Human Leukemia Cells Proliferation Rate Through Downregulation of Cdc25 and PP2A Activation. In: XLI Annual Meeting of SBBq, 2012, Foz do Iguaçu. Annual Meeting of the Brazilian Biochemistry and Molecular Biology Society (SBBq), 2012.

Projetos de pesquisa

• 2021 - Atual

  Pathway Improvement for Ethanol Production by Thermophilic Bacteria Involving Pyruvate Decarboxylase: Library Screening and Reintroduction into C. thermocellum, Descrição: Several lines of evidence suggesting that conversion of pyruvate to acetyl-CoA limits ethanol titer. In many thermophilic bacteria, including C. thermocellum, this conversion is mediated by the pyruvate ferredoxin oxidoreductase (PFOR) enzyme. To identify metabolic bottlenecks in enzymes from C. thermocellum, we replaced the T. saccharolyticum enzymes with C. thermocellum enzymes. Of the three enzymes we tested, AdhE (which provides both acetaldehyde dehydrogenase and alcohol dehydrogenase activity) and NfnAB (which transfers electrons from ferredoxin and NADH to NADP+) did not limit ethanol titer in T. saccharolyticum (ethanol production was > 60 g/L). However, Pfor reduced ethanol production by 7-fold, from 60 g/L to 9 g/L (Cui et al. in review). In a second line of evidence, expressing the T. saccharolyticum Pfor enzyme in C. thermocellum increased ethanol titer from 10 to 20 g/L (Hon et al. in review). Pyruvate decarboxylase (PDC) is an enzyme used by Saccharomyces cerevisiae and Zymomonas mobilis, two organisms known for their ability to produce ethanol at high titer (Olson et al., 2015). Although this enzyme has been successfully transferred to several mesophilic organisms, it does not seem to function well above 45°C in vivo. There have been several attempts to introduce pdc genes into C. thermocellum, however the maximum titer we achieved was 21 g/L, and the enzyme activity was 100-fold lower than what we observed in E. coli (0.3 U/mg vs 31 U/mg) (Tian et al., 2017). We think this problem is due to folding (unpublished data) and therefore the challenge is to increase the thermostability of the Pdc protein. We propose to build a single-site saturation mutagenesis library of the Acetobacter pasteurianus Pdc protein (10,621 members) using the CREATE system (Garst et al., 2016) to ensure even coverage of the experimental space. We will then screen this library in C. thermocellum for increased ethanol production. Promising mutations will also be used to develop computational models of protein stability. Milestones ? Generate CREATE library of Pdc protein mutants ? Improve system for screening Pdc mutants in C. thermocellum ? Screen 10,621 member Pdc mutant library in C. thermocellum for improved ethanol production ? Transfer 15 best candidate enzymes to high-ethanol-producing strain of C. thermocellum and measure ethanol yield and titer The chosen trainee will have some involvement with work pursuant to all of these milestones, with an anticipated lead role in library development and screening. The trainee will work as part of a 3-person team along with a Post Doc as well as a PhD student, both at CBMEG. The primary responsibility of the trainee will be to leverage the time of the Post Docs by attending to important but repetitive tasks such as running gels, preparing recombinant constructs, and sample analysis. As described in the proposal, this team will work be involved with Project i), Pathway Improvement, within Focus Area 2, Biotechnology, led by Dr, Dan Olson of Dartmouth. Situated within the multi-disciplinary A2G lab at Unicamp and working in close coordination with the research group led Professor Lynd at Dartmouth, the position offers outstanding opportunities for interdisciplinary and international research collaboration.. , Situação: Em andamento; Natureza: Pesquisa. , Integrantes: Milena Andreotti Minetto - Coordenador / Daniel G. Olson - Integrante / Sindelia Azzoni - Integrante / Lee Lynd - Integrante.

• 2017 - 2017

  Structural Study of Mur Enzymes Involved in the Biosynthesis of the Bacterial Wall, Descrição: Antibiotic resistance is a worldwide problem that calls for the employment of a variety of techniques for the identification of new antibacterial development targets. The bacterial cell wall is a complex structure that surrounds the entire cell and is essential for its survival, and the study of the macromolecular complexes that participate in its biosynthesis can open new avenues for the development of new antimicrobials. Mur enzymes are sequential proteins involved in cell wall formation, and their interaction is crucial for bacterial growth. To study the existence of a protein complex with the Mur proteins (MurEF, MurG and MurC), we use the bacterial two-hybrid technique (BACTH) and pull-down techniques of the three proteins in the E. coli operon. This project is complementary to the structural and functional studies performed in the group that aim towards the detailed characterization of this essential bacterial complex.. , Situação: Concluído; Natureza: Pesquisa. , Integrantes: Milena Andreotti Minetto - Coordenador / Andrea Dessen - Integrante / Viviana Job - Integrante., Financiador(es): Labex GRAL Master II Research Scholarship - Bolsa.

• 2016 - 2016

  Identification and Characterization of the Interaction Domain of the Factor H from the Complement System with the SdrE Protein of Staphylococcus aureus, Descrição: The pathogenic bacteria Staphylococcus aureus is the agent of respiratory and epithelial infections. During the first stages of infection, S. aureus resides and replicates inside the host cells and uses elaborate strategies to protect itself from the immune system. One of the strategies involves SdrE, a protein found on the bacteria surface that binds to the human factor H (fH), a regulator of the immune system. The project intended to identify and understand the interaction between SdrE from the S. aureus and the human factor H.. , Situação: Concluído; Natureza: Pesquisa. , Integrantes: Milena Andreotti Minetto - Integrante / Andrea Dessen - Coordenador / Daniel Maragno Trindade - Integrante.

• 2014 - 2014

  Kinetic of Proteins and mRNAs Expression of COUP-TFII and MyoD during the Myogenesis in C2C12 Cell Line, Descrição: The process of myoblast differentiation is regulated by hormonal stimulus that are, between others, activated by the MyoD transcription factor, which is, as studies suggest, necessary for the determination process. At the same time, COUP-TFII acts primarily in the differentiation process of myoblasts and osteoblasts, and as shown in previous studies, has an essential part in the commitment and destiny of the cell. The part COUP-TFII plays in the formation of skeletal muscle has not been clarified until the recent moment. The project had the objective to delineate kinetics of the repression of COUP-TFII mRNA and protein during the determination and differentiation of the C2C12 cell line during myogenisis.. , Situação: Concluído; Natureza: Pesquisa. , Integrantes: Milena Andreotti Minetto - Integrante / Henrique Marques-Souza - Coordenador / Carolina de Almeida Araujo - Integrante.

• 2011 - 2012

  Influence of Butyrate on Resistant Leukemia Cells Metabolism, Descrição: Chronic Myeloid Leukemia (CML), characterized mainly by the gene fusion BCR-ABL, still presents shortcomings in the treatment of patients. Butyrate has proven to be very effective in inducing tumor cell death primarily by inhibiting histone deacetylase and thus alter the expression of a variety of genes. However, little is known about the influence of butyrate on signaling pathways. Therefore, the main objective of the project was to evaluate the effect of butyrate on the activity and expression of the low molecular weight protein tyrosine phosphatase (LMWPTP) and P-glycoprotein (P-gp).. , Situação: Concluído; Natureza: Pesquisa. , Integrantes: Milena Andreotti Minetto - Integrante / Julia Laura Fernandes Abrantes - Integrante / Carmen Verissima Ferreira - Coordenador., Financiador(es): Fundação de Amparo à Pesquisa do Estado de São Paulo - Bolsa.

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