Yum D Y, Chung H C, Bai D H, Oh D H, Yu J H
Department of Food and Biotechnology, College of Engineering, Yonsei University, Seoul, Korea.
Biosci Biotechnol Biochem. 1994 Mar;58(3):470-4. doi: 10.1271/bbb.58.470.
SAP, an extracellular alkaline serine protease produced by Streptomyces sp. YSA-130, was purified to homogeneity by CM-Sephadex column chromatography and crystallization. The enzyme was a monomeric protein with a molecular weight of 19,000 as estimated by SDS-PAGE and gel filtration. The amino acid composition and amino-terminal sequence of SAP were similar to those of other bacterial serine proteases, i.e., Streptomyces griseus proteases A and B, Lysobacter enzymogenes alpha-lytic protease and Nocardiopsis dassonvillei subsp. prasina OPC-210 alkaline serine protease NDP-I. The optimum temperature and pH for the enzyme activity were 60 degrees C and 11.5. The enzyme was stable up 50 degrees C, and between pHs 4 and 12. The activity was inhibited by Ag+, Hg2+, Co2+, sodium dodecyl sulfate. N-bromosuccinimide, diisopropyl phosphorofluoridate (DFP), 2,3-butanedione, 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), iodoacetate, N-ethylmaleimide (NEM), phenylmethanesulfonyl fluoride (PMSF), and phenylglyoxal.
链霉菌属YSA - 130产生的细胞外碱性丝氨酸蛋白酶SAP，通过CM - 葡聚糖凝胶柱色谱法和结晶法纯化至同质。经SDS - PAGE和凝胶过滤估计，该酶为单体蛋白，分子量为19,000。SAP的氨基酸组成和氨基末端序列与其他细菌丝氨酸蛋白酶相似，即灰色链霉菌蛋白酶A和B、产酶溶杆菌α - 裂解蛋白酶以及达松维尔诺卡氏菌亚种普拉西纳OPC - 210碱性丝氨酸蛋白酶NDP - I。该酶活性的最适温度和pH分别为60℃和11.5。该酶在50℃以下以及pH值4至12之间稳定。其活性受到Ag +、Hg2 +、Co2 +、十二烷基硫酸钠、N - 溴代琥珀酰亚胺、二异丙基氟磷酸酯（DFP）、2,3 - 丁二酮、5,5'-二硫代双 -（2 - 硝基苯甲酸）（DTNB）、碘乙酸、N - 乙基马来酰亚胺（NEM）、苯甲基磺酰氟（PMSF）和苯乙二醛的抑制。
