Suryani Dyah Astuti, NIDN. 0008046902 and Welina Ratnayanti and Puspa Erawati and Djoni Izak Rudyardjo and Suhariningsih, NIDN. 0027065203 and Geoffrey Will (2011) Effect of Exogenous ALA to Increase the Production of Endogenous Porphyrin Staphylococcus aureus Bacteria For Applying Photodynamic Inactivation of Bacteria. In: Proceedings 3rd International Conference and Workshops on Basic and Applied Sciences. Enabling Research Innovation on Sciences and Technology to Meet Global Challenges. Faculty of Science and Technology Universitas Airlangga, Surabaya. ISBN 978-979-19096-1-7

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Abstract

Aminolevulinic acid (ALA) is a naturally occurring metabolite in the synthesis pathway of cellular heme production. In bacteria, addition of ALA may induce porphyrin synthesis. ALA induction leads to an increase in the synthesis of uroporphyrin, coproporphyrin and protoporphyrin IX, which are the immediate precursors of heme. In this research, the suspension of Gram positive Staphylococcus aureus bacteria ATCC 6538P was added with exogenous ALA (Sigma) in order to increase the production of bacteria endogenous porphyrin, with varying concentration (0, 1.5 mM, 2.5 mM, 3.5 mM, and 5 mM). Bacteria suspension was incubated in the dark at 37oC for 3 h for induction and production of endogenous porphyrin. Each extract, containing the produced endogenous porphyrins, characterized by fluorescence, UV-Visible spectroscopy and HPLC using reverse phase chromatography. Porphyrin production was demonstrated by the fluorescence emission peaks that appeared in the extracts of the ALA treated bacteria. The result showed that the excitation of the Staphylococcus aureus endogenous porphyrin at 405 nm yielded a fluorescence peak at 623 nm. The increasing of bacteria porphyrin absorbance comparable with increasing of ALA concentration. A significantly absorbance increased in the incubation of 3.5 mM ALA. The HPLC analyzed showed that the amount of porphyrin produced increased comparable with incubation of ALA concentration. A significantly endogenous porphyrin production increased in the incubation of 3.5 mM ALA. So the incubation of 3.5 mM ALA concentration significantly increased the bacteria endogenous porhyrin production and the absorption of porphyrin for applying the effectiveness photoinactivation of pathogen bacteria.

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