16th Brazilian Meeting on Organic Synthesis – 16th BMOS – November 15-18, 2015 - Búzios, Brazil

Endophytic fungi: Lipase Activity and Biotechnological

Applications

Karla S. C. da Rocha1*, Maria S. R. Queiroz1, Guilherme Maxwell1, Denise O.

Guimarães1, Ivaldo I. Junior2 e Ivana C. R. Leal1.

1Laboratório de Produtos Naturais e Ensaios Biológicos (LaProNEB)- CEP: 21941-902, Federal University of

Rio de Janeiro, Brazil. 2Department of Bioengineering, School of Chemistry - CEP: 21941-909 – Federal

University of Rio de Janeiro, Brazil.

* karlacotta@gmail.com

Keywords: endophytic fungi, lipases, monoacyglycerols

INTRODUCTION

Nowadays microbial lipases have great industrial importance. However, a few papers have explored the potential of endophytic fungi, micro-organisms that apparently not cause any damage to their host1, as a source of these enzymes. Our research group identified and isolated many endophytic fungi associated with some plant species, which may represent important sources of promising lipases. Monoacylglycerols (MAGs) are amphiphilic molecules that have applications such as emulsifiers, solubility agents and chiral building blocks. They are currently produced by alkaline glycerolysis oil under high P and T, resulting in low yields and numerous by-products. The use of enzymatic processes catalyzed by lipase, may lead to a more ecological approach2. This work aimed to investigate lipase activity of endophytic fungi as well as their application as biocatalysts in esterification reactions for esters production.

RESULTS AND DISCUSSION

An initial screening of lipase activity was initially held by the Rhodamine B (0,001%) method, at 365 nm of absorbance. The HB13 strain presented fluorescence halo after 24h of inoculation, confirming the lipase activity. Lipolytic fungi were inoculated into a pre-fermentation medium and incubated for 24 h at 30 °C, and then transferred into a fermentation medium to stimulate the production of lipase, incubated for 48h, 30°C and 180rpm of stirring. From the fermentation medium was performed palmitate hydrolysis of p-nitrophenyl method (pPNN). The enzyme activity was defined as the concentration of p-nitrophenol in mol/L, based on the molar absorption coefficient of p-nitrophenol obtained from a calibration curve (R2 = 0.97) at 410

nm. The best result was obtained in 48h of incubation with the HB13 and TB1fungus (Graphic 1).

Figure 1. Medium with HB13 fungus containing Rhodamine B fluorescing halo.

Graphic1. p-nitrophenol concentration: HB13 (92U/mL);

TB1 (85U/ml); TB2 (5U/mL).

CONCLUSION

For the assays Rhodamine B and pPNN TB1 and HB13 strain were promising for lipase activity. Promising species will be investigated for the esterification of activity, and will later be submitted to lipase and MAGs production reactions.

ACKNOWLEDGEMENTS

CAPES, CNPq, FINEP and FAPERJ.

REFERENCES

1. SCHULZ, B.; BOYLE, C. (2005). Mycol Res 109:661–686; 2. JUNIOR, I.I.;

FLORES, M.C.; SUTILI, F.K.; LEITE, S.G.F.; MIRANDA, L.S.; LEAL, I.C.R. DE SOUZA, R.O.M.A. (2012). Journal of Mol Cat. B, Enz., 77:53– 58.

85 U/ml

5 U/ml

92 U/ml