Purification and Properties of Alkaline Lipase from Pseudomonas sp. J-19

Pseudomonas sp. J-19가 생산하는 Alkaline Lipase의 정제와 특성

A strain J-19 was isolated from soil, produced lipase which has resistant against alkali and linear alkylbenzene sulfonate. The strain was identified as Pseudornonns sp.. The enzyme was purified by ammonium sulfate precipitation, DEAE-Sephadex and Sephadex G- 100 column chromatography. The specific activity of the purified enzyme was 35 unit/mg protein and the yield of enzyme activity was 17%. The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis. Mo1ecul;tr weight of the purified enzyme was estimated about 36,000 by Sephadex GI00 gel filtration and SDS-polyacrylarnide gel electrophoresis. The optimum pH and temperature were pH 10.0 and $30^{\circ}C$, respectively. Activity of the purified enzyme was increased 2-fold by the addition of 0.1% linear alkylbenzene sulfonate and 2.5- fold by the addition of 0.05% Tide. This enzyme remained stable from pH 8.0 to 10.0 and stable up to $40^{\circ}C$.

토양으로부터 분리한 알칼리 내성 및 liner alkylbenzene sulfonate 내성인 lipase 생산균주를 동정하여 Pseudomonas sp. J-19로 명명하였다. 호알칼리성 lipase는 ammonium sulfate 침전, DEAE-Sephadex와 Sephadex G-100 column chromatography로 정제하였고, 정제효소의 비활성도는 35 unit/mg protein, 수율은 17이었다. 정제효소는 polyacrylamide disc gel 전기영동에서 단일 band를 나타내었고, Sephadex G-100 gel filtration과 SDS-polyacrylamide gel 전기영동에 의하여 추정된 분자량은 36,000이었다. 정제효소의 최적 pH는 10.0 최적온도는 30'C이었다. 정제 효소의 활성은 0.1 linear alkylbenzene sulfonate 첨가에 의하여 2배 증가되었고, 0.05 Tide에 의하여 2.5배 증가되었다. 정제효소는 p8.0-10.0, 4'C이하에서 안정하였다.