Millson Stefan H, Evans Ivor Howell
Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
Antonie Van Leeuwenhoek. 2007 Nov;92(4):399-404. doi: 10.1007/s10482-007-9168-0. Epub 2007 Jun 9.
The soil yeast Lipomyces starkeyi (NCYC 1436) secretes dextranase activity into the growth medium. Resolution of a dextranase-active protein fraction by SDS-PAGE produced three protein bands, of 66 kDa, 68 kDa and 78 kDa, and isoelectric focusing of the same fraction resulted in seven protein bands, of pIs 3.50, 3.85, 4.20, 4.80, 4.85, 5.00 and 5.30. Dextranase activity was demonstrated for all the isoelectric forms, and for the 78 kDa species in the presence of SDS. Amino acid compositions of the 66 kDa, 68 kDa and 78 kDa protein bands were determined, and the N-termini of the 66 kDa and 78 kDa protein bands were sequenced: the first two amino acids at the N-terminus of each protein were alanine and valine, respectively; an alanine-valine pair is seen early in the N-terminal coding sequences of the dextranases and the isopullulanase produced by the phylogenetically disparate organisms contributing to glycosyl hydrolase family 49.
土壤酵母斯达氏油脂酵母(NCYC 1436)将葡聚糖酶活性分泌到生长培养基中。通过SDS-PAGE对具有葡聚糖酶活性的蛋白组分进行分离,产生了三条蛋白带,分子量分别为66 kDa、68 kDa和78 kDa,对同一组分进行等电聚焦则得到七条蛋白带,其等电点分别为3.50、3.85、4.20、4.80、4.85、5.00和5.30。所有等电形式的蛋白均表现出葡聚糖酶活性,在SDS存在的情况下,78 kDa的蛋白也具有该活性。测定了66 kDa、68 kDa和78 kDa蛋白带的氨基酸组成,并对66 kDa和78 kDa蛋白带的N端进行了测序:每种蛋白N端的前两个氨基酸分别为丙氨酸和缬氨酸;在属于糖基水解酶家族49的系统发育不同的生物体产生的葡聚糖酶和异普鲁兰酶的N端编码序列中,早期可见丙氨酸-缬氨酸对。