Deteksi dan Analisis Gen NRPS-PKS Pada Aktinomiset Penghasil Biopigmen

Abstract

Biopigmen yang dihasilkan mikrob terutama aktinomisetes memiliki beragam fungsi seperti antioksidan, antimikrob, dan antikanker. Biopigmen merupakan metabolit sekunder, beragam metabolit sekunder dihasilkan melalui jalur sintesis protein non ribosomal dan poliketida sintetase (NRPS-PKS). Penelitian ini bertujuan untuk menganalisis domain adenilasi (A) dari NRPS dan domain ketosintase (KS) dari PKS pada aktinomiset penghasil biopigmen. Terdapat lima isolat yang digunakan dalam penelitian ini yaitu HV 4, HV 11, JSN 1.9, JSN 28, dan NTB 42. Genom dari kelima isolat diekstrasi, domain A diamplifikasi dan diklon, kemudian hasil klon di sekuen dan dilakukan analisis bioinformatika. JSN 1.9, JSN 28, dan NTB 42 memiliki amplikon domain A yang memiliki ukuran ±750 pb. Hasil alignment domain A dari JSN 1.9, JSN 28, dan NTB 42 menunjukkan bahwa terdapat 208 nukleotida yang termasuk ke dalam daerah terkonservasi. Model protein dibuat dari hasil sekuen ketiga isolat dan menunjukkan kualitas model yang baik. Dari lima isolat domain A hanya ditemukan pada tiga isolat.

Bio-pigments produced by microbes especially actinomycetes have shown various function such as antioxidant, antimicrobe, and anticancer. Bio-pigments are secondary metabolite, a wide variety of them are produced through non ribosomal protein and polyketide synthase (NRPS-PKS) pathway. This study aims to detect and analyze adenylation (A) domain from NRPS and ketosynthase (KS) domain PKS in biopigment producing bacteria. There are five isolates used in this research, which are HV 4, HV 11, JSN 1.9, JSN 28, and NTB 42. Genome was extracted from the isolates, A domain gene was amplified and cloned. Furthermore, recombinant plasmids were sequenced and bioinformatic analysis was done. A domain amplicon was found in JSN 1.9, JSN 28, and NTB 42, with the size of ±750 bp. JSN 1.9, JSN 28, and NTB 42 adenylation domain alignment result shows that there are 208 nucleotides belong to conserved region. Protein models constructed from three sequenced adenylation domain are predicted to have good qualities and displayed behavior similar to that in nature. From five isolates A domain was only found in three isolates.