PEMURNIAN DAN KARAKTERISASI ENZIM PGA INTRASEL DARI BACILLUS SP. BAC4
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...
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| Format: | Theses |
| Language: | Indonesia |
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| Online Access: | https://digilib.itb.ac.id/gdl/view/1776 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | 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. |
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