rd
23 Congress of the International Union for Biochemistry and Molecular Biology
th
44 Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology
th
th
Foz do Iguaçu, PR, Brazil, August 24 to 28 , 2015
PROTEIN PROFILE OF Fusarium oxysporum AND Phanerochaete chrysosporium
DURING THE LAPACHOL BIOTRANSFORMATION PROCESS.
Coitinho, L.B.1; Siqueira A.C.R1; Emery, F.S.1; Cabral, H.1
1
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo.
Biotransformation is an important process in enzyme related chemical transformations.
Fungi can be considered as a promising source of new compounds, mainly for chiral
reactions. Chemo-, regio-, and stereoselective processes are very important in the
synthesis of many chemical and pharmaceutical intermediates. Lapachol is a
naphthoquinone that presents many interesting biological activities, which have
potential uses in the pharmaceutical industry. The aims of this project are to analyze the
protein profile of Fusarium oxysporum and Phanerochaete chrysosporium during the
lapachol biotransformation process. The microorganism was cultivated in 250 mL
Erlenmeyer flasks containing 50 mL of soy-glucose medium and incubated on a rotary
shaker at 120 rpm at 30°C. After 24 hours, the mycelial mass was transferred to a
Czapeck medium containing lapachol solution in DMSO (final concentration of 50
µg/mL) and fermentation was continued for 24 and 48 hours. The secretome and
proteome were obtained through precipitation followed by SDS-Page 12%. In addition,
the proteolytic and lipase activities were determined using fluorescenceresonance
energy transfer (FRET) substrate and 4-nitrophenyl palmitate, respectively. The
obtained results suggest that lapachol modifies the profile of the proteins production
compared to the control. Both proteomes of P. chrysosporium and F. oxysporum
presented protease activity, although both controls showed a higher production in
comparison to the fermentation with lapachol. Moreover, both secretome during the
biotransformation process did not show any protease activity. The proteome and
secretome of P. chrysosporium in the presence of lapachol did not show lipase activity
whereas F. oxysporum showed lipase activity. According to the results, the fungi modify
their protein profile when exposed to lapachol during the biotransformation process.
Acknowledgements: Fundação de Amparo à Pesquisa do Estado de São Paulo
(FAPESP). Processo Nº 2014/01497-9 e 2012/24703-8
Key Words: Biotransformation; Lapachol; Proteins.
Brazilian Society for Biochemistry and
Molecular Biology (SBBq)