Purification and Properties of Isocitrate Lyase from Saccharomycopsis lipolytica

Saccharomycopsis lipolytica Isocitrate Lyase의 정제와 성질

Isocitrate lyase from crude extract of Saccharomycopsis lipolytica ATCC44601 and MX9-11RX8 temperature-sensitive mutant was purified about 54 times and 87 times, respectively by ammonium sulfate fractionation, Toyo peal HW-55F gel filtration and DEAE-Cellulose ion exchange chromatography, The molecular weight of the purified isocitrate lyase from this yeast was estimated to be 230, 000 by gel filtration on Sephadex G-200, and SDS-polyacrylamide Eel electrophoresis showed that the enzyme consisted of four identical or similar subunits with a molecular weight of 59, 000 and the enzyme showed optimum activity at pH 6.9.

Saccharomycopsis lipolytica ATCC 44601 과 MX9-11RX8 온도감수성 변이균주의 isocitrate lyase는 조추출액을 ammonium sulfate 분획, Toyo Peal HW-55F gel filtration, DEAE-Cellulose ion exchange chromatography 등의 방법에 의하여 각각 54배, 87배 분리 정제되었다. 정제효소의 subunit 분자량은 59,000이고 Sephadex G-200 gel filtration에 의한 native enzyme 은 230,000이므로 이 효모의 isocitrate lyase는 같거나 비슷한 subunit 4개로 구성된 tetramer이며, 최적 pH는 6.9이었다.