Penghasilan enzim tanase oleh penicillium sp. pencilan tempatan secara fermentasi kultur tenggelam
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Date
2007-03
Authors
Sandal, Doblin Anak
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Abstract
The isolate that was used in the production of tannase was identified as Penicillium
sp. This isolate was isolated from a R. apiculata barks dumping area, at the
mangrove area in Larut-Matang, Perak. Optimization of physical parameters and
medium compositions showed an increment of 578% or 4.983 U/ml compared with
the activity before optimization which was 0. 735 U/ml. The optimal physical
parameters were initial pH 6.0, temperature of 30°C, agitation speed of 170 rpm
and inoculum size of 1.0% (v/v) of 1.53 x 106 spores per mi. Meanwhile, optimal
medium compositions were {%; w/v) NHCI4, 0.25%; KH2P04, 0.1 %; MgS04.yH20,
0.05%; KCI, 0.05% and tannic acid, 4%. Cell immobilization of Penicillium sp. on
sodium alginate beads and nylon sponge cubes produced tannase activity as much
as 7.066 U/ml and 5.124 U/ml respectively. The results showed an increment of
44.2% for calcium alginate beads and 4.57% for nylon sponge cubes. Production
of tannase by free cells in a tubular air-lift fermenter showed an increment of 71.7%
compared to Erlenmeyer shake flask volume 500 ml and 73.5% compared to
Erlenmeyer shake flask volume 250 mi. Maximal production of tannase in a tubular
air-lift fermenter was 11.76 U/ml with increment of 38.4% before optimization. The
optimized production parameters used in the fermenter were 400 of sodium
alginate beads, inoculum size at 1.0% (v/v) of 1.53 x 106 spores per ml and
aeration of 2.0 wm. Tannase was purified by ammonium sulphate precipitation and
gel filtration chromatography using Sephadex G-200, twice. The peak obtained
was purified about 44.35 fold with a yield of 0.69% and specific activity of 6.12
U/mg protein. Its molecular weight was estimated to be around 76,1000 Dalton by
SDS-PAGE. Crude and purified tannase had the optimum temperature of 40°C and
stable at that temperature for 90 minutes and 75 minutes, respectively.
Description
Keywords
Enzim tanase , Penicillium sp. pencilan , Fermentasi kultur tenggelam