该文对一种黑曲霉来源的蛋白酶CpyC进行了生化特征解析。该酶在结构与组成上具有丝氨酸蛋白水解酶SB家族特征性催化活性中心,由Asp188、His226和Ser381氨基酸残基组成,序列特征接近丝氨酸蛋白水解酶SB家族S08类,但与现有家族成员分类特征并不完全相同。重组CpyC的最适作用温度和pH值分别为50 ℃和8.0,Ca2+和Co2+对其分别具有最强的激活作用和抑制作用,丝氨酸蛋白酶特异性抑制剂苯甲基磺酰氟对其具有较强抑制作用,可见,黑曲霉蛋白酶CpyC是一种碱性丝氨酸蛋白酶。CpyC水解大豆分离蛋白结果显示,该酶具有水解蛋白质为多肽和寡肽的活性特征,在蛋白质生物加工中可能具有潜在应用价值。
A proteolytic enzyme, CpyC, from Aspergillus niger was biochemically characterized.It contained a typical catalytic active site belonging to that of serine peptidase SB clan, with corresponding conserved amino acid residues Asp188, His226, and Ser381.Its conserved sequence or regions were similar to that of S08 class of the serine peptidase SB family, however, not completely identical to the existing family members' classification features.The purified recombinant CpyC showed its maximum activity at 50 ℃ and pH 8.0.The metal ion, Ca2+, showed the strongest activation effects on CpyC while Co2+ had the strongest inhibition effects.Its activity was strongly inhibited by phenylmethylsulfonyl fluoride, a specific serine protease inhibitor.The results suggested that protease CpyC from A.niger is an alkaline serine peptidase.CpyC could hydrolyze soybean protein isolate effectively and formed the main hydrolytes with polypeptide and oligopeptide, promoting its potential applications in protein bioprocessing.
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