Section Molecular Genetics, Department of Genetics, Agricultural University Wageningen, Dreyenlaan 2, 6703 HA Wageningen, and Laboratory for Electron Microscopy, Biological Center, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands.
Appl Environ Microbiol. 1992 Apr;58(4):1190-4. doi: 10.1128/aem.58.4.1190-1194.1992.
The subcellular localization of glucose oxidase (EC 1.1.3.4) in Aspergillus niger N400 (CBS 120.49) was investigated by (immuno)cytochemical methods. By these methods, the bulk of the enzyme was found to be localized in the cell wall. In addition, four different catalases (EC 1.11.1.6) were demonstrated by nondenaturing polyacrylamide gel electrophoresis of crude extracts of induced and noninduced cells. Comparison of both protoplast and mycelial extracts indicated that, of two constitutive catalases, one is located outside the cell membrane whereas the other is intracellular. Parallel with the induction of glucose oxidase, two other catalases are also induced, one located intracellularly and one located extracellularly. Furthermore, lactonase (EC 3.1.1.17) activity, catalyzing the hydrolysis of glucono-delta-lactone to gluconic acid, was found to be exclusively located outside the cell membrane, indicating that gluconate formation in A. niger occurs extracellularly.
通过(免疫)细胞化学方法研究了黑曲霉 N400(CBS 120.49)中葡萄糖氧化酶(EC 1.1.3.4)的亚细胞定位。通过这些方法,发现大部分酶定位于细胞壁中。此外,通过诱导和非诱导细胞的粗提物的非变性聚丙烯酰胺凝胶电泳,还证明了四种不同的过氧化氢酶(EC 1.11.1.6)。比较原生质体和菌丝体提取物表明,两种组成型过氧化氢酶之一位于细胞膜外,另一种位于细胞内。与葡萄糖氧化酶的诱导平行,还诱导了另外两种过氧化氢酶,一种位于细胞内,另一种位于细胞外。此外,还发现了内酯酶(EC 3.1.1.17)的活性,该酶催化葡萄糖酸-delta-内酯水解生成葡萄糖酸,该酶仅位于细胞膜外,表明黑曲霉中葡萄糖酸盐的形成发生在细胞外。