PEMURNIAN DAN KARAKTERISASI ENZIM PGA INTRASEL DARI BACILLUS SP. BAC4

• Main Author: BAHRI, SYAIFUL
• Format: Theses
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/1776
• Institution: Institut Teknologi Bandung

A l ocal strain of Bacillus sp. BAC4 has been identified to produce <br /> penicillin G acylase (PGA). This enzyme hydrolyses benzylpenicillin to <br /> 6-aminopencillanic acid and phenylacetic acid. The 6-aminopenicillanic <br /> acid is a valuable starting material for the production of various <br /> derivatives of a-laktam antibiotics. Previous research had shown that <br /> Bacillus sp. BAC4 able to produce both extracellular and intracellular <br /> PGA. The extracellular PGA of Bacillus sp. BAC4 has been studied <br /> extensively, but the intracellular PGA has not been characterized. This <br /> research was aimed to study the properties of intracellular PGA, through <br /> purification and characterization. Intracellular PGA purification was <br /> done by sonication, followed by centifugation to sparated the filtrate <br /> from cell debris. The filtrate was than fractionated by 80 % (w/v) <br /> ammonium sulphate, dialysed in selophan tube, and fractionated by <br /> affinity chromatography using phenylacetate ligands. <br /> Three fractions <br /> were obtained, two of which showed protease activity whilst the other <br /> one has acylase activity. <br /> Acylase fraction contained a protein of 32.5 <br /> mg with a specific activity of 0.29 unit/mg. Further characterization by <br /> SDS-PAGE showed two distinct bands at 47 kD and 35 kD. This data is <br /> in in good agreement with the intracellular PGA of L coli, K. <br /> citrophilia, A. viscocus and P. rettgeri which also contained of two subunit <br /> moleculs with a relative molecular mass of 80 kD. <br /> This low <br /> molecular weight was unexpected, as the extracellular PGA of Bacillus <br /> sp. BAC4 having a larger molecular mass of 130 M This result <br /> suggest that the intracellular and the extracellular PGA of Bacillus sp. <br /> BAC4 might be of two different uncorrelated molecules.