摘要
采用硫酸铵分段盐析、透析、DEAE-纤维素离子交换层析和葡聚糖Sephadex G-100凝胶层析,从南日鲍内脏中分离纯化得到一种β-葡萄糖苷酶.SDS-PAGE分析表明,所纯化的β-葡萄糖苷酶为单聚体,分子量约为90 kDa,N-端8个氨基酸残基序列为AGMLYPRV.β-葡萄糖苷酶的最适温度为50℃,最适pH值为5.0,Mn2+、Cu2+、Ba2+和Zn2+对酶活力有显著的促进作用,而Al3+对酶有明显的抑制作用,SDS则使酶变性失活;pNPG与β-葡萄糖苷酶之间的亲和程度最高,纤维二糖和京尼平苷次之,水杨苷最差.
Theβ -glucosidase from abalone （Haliotis diversicolor diversicolor） visceral was separated, purified through a combined technique including precipitation with ammonium sulfate, dialysis, ion - exchange chromatography and gel - filtration chromatography. The result of SDS - PAGE showed that the enzyme is a single dimer, which has a molecular weight of about 90 kDa, as well as the 8 N - ter- minal amino acids sequence is AGMLYPRV. The optimum temperature of the β- glucosidase was 50 ℃,, and the pH was 5.0. The enzyme activity was apparently promoted by Mn2＋ , Cu2＋ , Ba2＋ and Zn2＋ but strongly inhibited by Aβ ＋ , and SDS will inactivate the enzyme completely. The experimental data also indicated that pNPG was the best substrate amomg the four substrates, including cellobiose, geniposide and salicin.
出处
《福州大学学报（自然科学版）》
CAS
CSCD
北大核心
2014年第3期480-485,共6页
Journal of Fuzhou University(Natural Science Edition)
基金
国家海洋公益性行业科研专项子课题(201205022-3)
福建省海洋渔业厅重点项目(闽海渔合同2010〗2-27号)
关键词
南日鲍
内脏
Β-葡萄糖苷酶
纯化
酶学性质
Haliotis discus nanri
viscera
β- glucosidase
purification
properties
