Screening for genomic sequences which codify lipolytic enzymes specialized in petroleum hydrocarbons degradation.

Author(s)

Thais Carvalho Maester

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI)
• Defense date: 2011-05-30

Examining board members

Eliana Gertrudes Macedo Lemos; Luiz Juliano Neto; Tsai Siu Mui

• Advisor: Eliana Gertrudes Macedo Lemos

Abstract

Lipolytic enzymes have show enormous biotechnological potential. The work was done to find genes which codify lipolytic enzymes in a metagenomic library with 4224 clones. Clones were selected according to lipolytic activity and were assessed by cultivation in medium supplemented with tributyrin. Assessment was done by observation of halos formed around the colonies, with 30 clones producing halos. Of these, two were selected. DNA from the sub libraries was sequenced, generating a complete contig for clone PL28.F10 that was compared to sequences from the NCBI. An ORF of 303 amino acids with 61% of identity with uncunturable microorganism were found. The clone presented the ORF15 similar to that of lipolytic enzyme family IV. The alignments made possible the identification of active sites which represent the family, confirming the results obtained with the construction of the cladograms. The ORF15 showed similarities to patented BASF esterase/lipase and an unnamed protein of CAMBIA. (AU)

• FAPESP's process: 09/03772-9 - Characterization of agarase activity from Burkholderia sp.
• Grantee: Thaís Carvalho Maester Casanova
• Support Opportunities: Scholarships in Brazil - Master