Electronic Theses and Dissertation

Abstrak pseudomonas aeruginosa merupakan bakteri oportunistik yang mampu membentuk biofilm sehingga berperan penting dalam peningkatan virulensi dan resistensi terhadap antibiotik. pembentukan biofilm dikendalikan oleh berbagai gen regulator dan struktural, di antaranya lasr sebagai regulator quorum sensing serta psla dan pela sebagai gen struktural penyusun matriks ekstraseluler (eps). kopi arabika gayo (coffea arabica l.) diketahui mengandung berbagai senyawa bioaktif yang berpotensi sebagai antibiofilm. penelitian ini bertujuan untuk melihat aktivitas antibiofilm ekstrak kopi arabika gayo terhadap p. aeruginosa atcc 27853 sebagai dasar pendekatan molekuler terhadap ekspresi gen lasr, psla, dan pela yang berperan dalam pembentukan biofilm p. aeruginosa. penelitian ini merupakan penelitian eksperimental laboratorik menggunakan ekstrak biji kopi arabika gayo yang diperoleh melalui metode maserasi dengan pelarut etanol 96%. komposisi bioaktif ekstrak dianalisis dengan uji fitokimia dan gas chromatography-mass spectrometry (gc-ms). aktivitas antibiofilm diamati melalui uji hambat serta degradasi biofilm pada inkubasi 24 dan 48 jam pada konsentrasi 0,25% dan 0,125%. rna hasil kultur diekstraksi dan disintesis menjadi cdna menggunakan kit komersial. optimasi amplifikasi gen lasr, psla, dan pela dilakukan dengan polymerase chain reaction (pcr) konvensional. analisis ekspresi gen lasr, psla, dan pela direncanakan menggunakan metode quantitative pcr, namun belum dapat dilakukan karena keterbatasan teknis. ekstrak kopi arabika gayo mengandung berbagai senyawa bioaktif hasil analisis gc-ms, antara lain caffeine, 2-furanmethanol, desulphosinigrin, dan senyawa bioaktif turunan lainnya. pengujian aktivitas antibiofilm menunjukkan bahwa konsentrasi 0,25% dan 0,125% mampu menghambat dan mendegradasi biofilm p. aeruginosa. pada uji degradasi biofilm, konsentrasi paling efektif pada inkubasi 24 jam adalah 0,25%, sedangkan pada inkubasi 48 jam adalah 0,125%. optimasi pcr gen lasr, psla, dan pela telah berhasil dilakukan dan menghasilkan produk berukuran 130 bp, 149 bp, dan 119 bp. kata kunci: kopi arabika gayo, pseudomonas aeruginosa, biofilm, antibiofilm, gc-ms

ABSTRACT Pseudomonas aeruginosa is an opportunistic bacterium capable of forming biofilms, thus playing an important role in increasing virulence and resistance to antibiotics. Biofilm formation is controlled by various regulatory and structural genes, including lasR as a Quorum Sensing regulator and pslA and pelA as structural genes that compose the extracellular matrix (EPS). Gayo Arabica Coffee (Coffea arabica L.) is known to contain various bioactive compounds that have the potential as antibiofilms. This study aims to observe the antibiofilm activity of Gayo Arabica Coffee extract against P. aeruginosa ATCC 27853 as a basis for a molecular approach to the expression of lasR, pslA, and pelA genes that play a role in the formation of P. aeruginosa biofilms. This study is a laboratory experimental study using Gayo Arabica Coffee bean extract obtained through the maceration method with 96% ethanol solvent. The bioactive composition of the extract was analyzed by phytochemical tests and Gas Chromatography-Mass Spectrometry (GC-MS). Antibiofilm activity was observed through inhibition tests and biofilm degradation at 24 and 48 hours of incubation at concentrations of 0.25% and 0.125%. RNA from the culture was extracted and synthesized into cDNA using a commercial kit. Optimization of lasR, pslA, and pelA gene amplification was carried out using conventional Polymerase Chain Reaction (PCR). Analysis of lasR, pslA, and pelA gene expression was planned to use the Quantitative PCR method, but could not be carried out due to technical limitations. Gayo Arabica Coffee extract contains various bioactive compounds as a result of GC-MS analysis, including caffeine, 2-furanmethanol, desulphosinigrin, and other derivative bioactive compounds. Antibiofilm activity testing showed that concentrations of 0.25% and 0.125% were able to inhibit and degrade P. aeruginosa biofilms. In the biofilm degradation test, the most effective concentration after 24 hours of incubation was 0.25%, while after 48 hours it was 0.125%. PCR optimization of the lasR, pslA, and pelA genes was successful, yielding products measuring 130 bp, 149 bp, and 119 bp, respectively. Keywords: Gayo Arabica coffee, Pseudomonas aeruginosa, biofilm, antibiofilm, GC-MS