Doblin Anak, Sandal
(2017)
Penghasilan enzim tanase oleh penicillium sp. pencilan tempatan secara fermentasi kultur tenggelam.
PhD thesis, Universiti Sains Malaysia.
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.
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