BIOKONVERSI SEFALOSPORIN C MENJADI ASAM 7-AMINOSEFALOSPORANAT DENGAN SEFALOSPORIN ASILASE
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Abstract
Cephalosporins are the most widely used class of β-lactam antibiotic in the world and clinically active against gram positive and gram negative bacteria. Cephalosporin C (CPC) is naturally produced by fungus Cephalosporiun acremonium. CPC has moderate antibacterial activity with minimum inhibitory concentration values of 25-100 µg/mL and 12-25 µg/mL for gram-positive and for gram-negative bacteria, respectively. CPC can be converted into 7-aminocephalosporonic acid (7-ACA) as intermediate compound for cephalosporin derivatives by two-steps or one-step enzymatic method. Two-step enzymatic method uses D-amino acid oxidase (DAAO) to produce glutaryl-7-amino cephalosporanic acid (GL 7-ACA) for the first step and GL-7-ACA acylase to produce 7-ACA for the second step. One-step enzymatic method uses CPC acylase to convert CPC into 7-ACA directly. Some microorganisms produce CPC acylase, such as Pseudomonas sp., Bacillus megaterium, Aeromonas sp., dan Arthrobacler. A natural CPC acylase has low activity and genetic engineering was used to increase its activity.
Keywords: Cephalosporin, cephalosporin acylase, 7-ACA, genetic engineering, mutation
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ABSTRAK
Sefalosporin merupakan antibiotik golongan β-laktam yang paling banyak digunakan di dunia dan secara klinis aktif terhadap bakteri gram positif dan gram negatif. Sefalosporin C merupakan sefalosporin alami yang dihasilkan oleh kapang Cephalosporium acremonium. Sefalosporin C mempunyai aktivitas antibakteri moderat dengan nilai konsentrasi hambat minimum 25-100 µg/mL untuk bakteri gram positif dan 12-25 µg/mL untuk bakteri gram negatif. Sefalosporin C dapat diubah menjadi asam 7-aminosefalosporanat (7-ACA) sebagai senyawa antara untuk pembuatan turunan sefalosporin dengan metode enzimatik secara dua atau satu tahap. Produksi 7-ACA secara enzimatik dapat menggunakan metode dua tahap dan satu tahap enzimatik. Metode enzimatik secara dua tahap menggunakan enzim asam D-amino oksidase (DAAO) untuk menghasilkan asam glutaril-7-aminosefalosporinat (GL-7-ACA) pada tahap pertama dan menggunakan asam glutaril-7-aminosefalosporinat asilase untuk menghasilkan 7-ACA pada tahap kedua. Metode enzimatik secara satu tahap menggunakan sefalosporin asilase untuk mengubah CPC menjadi 7-ACA secara langsung. Beberapa mikroorganisme penghasil sefalosporin asilase yaitu Pseudomonas sp., Bacillus megaterium, Aeromonas sp., dan Arthrobacter. Aktivitas CPC asilase alami sangat rendah dan rekayasa genetik digunakan untuk meningkatkan aktivitasnya.
Kata kunci : Sefalosporin, sefalosporin asilase, 7-ACA, rekayasa genetik, mutasiArticle Details
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