Baratto César Milton, da Silva Marcia Vanusa, Santi Lucélia, Passaglia Luciane, Schrank Irene Silveira, Vainstein Marilene Henning, Schrank Augusto
Centro de Biotechnologia, Universidade Federal do Rio Grande do Sul, UFRGS, Porto Alegre, RS, Brazil.
Can J Microbiol. 2003 Nov;49(11):723-6. doi: 10.1139/w03-085.
Albeit Metarhizium anisopliae is the best-characterized entomopathogenic fungus, the role of some hydrolytic enzymes during host cuticle penetration has not yet been established. Three chitinase genes (chit1, chi2, chi3) from Metarhizium have already been isolated. To characterize the chitinase coded by the chit1 gene, we expressed the active protein (CHIT42) in Escherichia coli using a T7-based promoter expression vector. The recombinant protein, CHIT42, is active against glycol chitin and synthetic N-acetylglucosamine (GlcNAc) dimer and tetramer substrates. These activities suggest that the recombinant CHIT42 acts as an endochitinase.
尽管绿僵菌是特征最明确的昆虫病原真菌，但一些水解酶在穿透寄主表皮过程中的作用尚未明确。已从绿僵菌中分离出三个几丁质酶基因（chit1、chi2、chi3）。为了表征chit1基因编码的几丁质酶，我们使用基于T7的启动子表达载体在大肠杆菌中表达了活性蛋白（CHIT42）。重组蛋白CHIT42对乙二醇几丁质以及合成的N-乙酰葡糖胺（GlcNAc）二聚体和四聚体底物具有活性。这些活性表明重组CHIT42作为一种内切几丁质酶发挥作用。
