den Herder I F, Rosell A M, van Zuilen C M, Punt P J, van den Hondel C A
TNO Medical Biological Laboratory, AA Rijswijk, The Netherlands.
Mol Gen Genet. 1992 Jun;233(3):404-10. doi: 10.1007/BF00265437.
An enzyme with alpha-galactosidase activity and an apparent molecular weight of 82 kDa was purified from culture medium of Aspergillus niger. The N-terminal amino acid sequence of the purified protein shows similarity to the N-terminal amino acid sequence of alpha-galactosidases from several other organisms. Oligonucleotides, based on the N-terminal amino acid sequence, were used as probes to clone the corresponding gene from a lambda EMBL3 gene library of A. niger. The cloned gene (aglA) was shown to be functional by demonstrating that the 82 kDa alpha-galactosidase is absent from a strain with a disruption of the aglA gene, and is over-produced in strains containing multiple copies of the aglA gene. Enzyme activity assays revealed that the 82 kDa alpha-galactosidase A represents a minor extracellular alpha-galactosidase activity in A. niger.
从黑曲霉培养基中纯化出一种具有α-半乳糖苷酶活性、表观分子量为82 kDa的酶。纯化蛋白的N端氨基酸序列与其他几种生物的α-半乳糖苷酶的N端氨基酸序列相似。基于该N端氨基酸序列的寡核苷酸被用作探针，从黑曲霉的λEMBL3基因文库中克隆相应基因。通过证明aglA基因被破坏的菌株中不存在82 kDa的α-半乳糖苷酶，而在含有多个aglA基因拷贝的菌株中该酶过量产生，表明克隆的基因（aglA）具有功能。酶活性测定表明，82 kDa的α-半乳糖苷酶A在黑曲霉中代表一种次要的细胞外α-半乳糖苷酶活性。
