Key Laboratory of Industrial Microbiology & Fermentation Technology of Yunnan Province, Yunnan University, Kunming 650091, Yunnan, P.R. of China.
Mycologia. 2004 Jan-Feb;96(1):16-22. doi: 10.1080/15572536.2005.11832991.
A neutral serine protease (designated Aoz1) was purified to homogeneity from a strain of Arthrobotrys oligospora, obtained from soil in Yunnan Province. The purified protein showed a molecular mass of approximately 38 000 Dalton, pI 4.9 and displayed optimal activity at 45 C and pH 6-8. The protein could hydrolyze gelatin, casein and the chromogenic substrate azocoll, and it could immobilize nematodes in vitro (Panagrellus redivivus L. [Goodey]). The level of activity in culture medium was found to increase with increasing gelatin concentration. Scanning electron micrographs demonstrated dramatic structural changes in nematode cuticle treated with the purified protease. A partial peptide sequence obtained by N-terminal sequence analysis was used to design degenerate primers for the isolation of a cDNA gene encoding the mature protease. Analysis of the cDNA and corresponding genomic sequence revealed 97% identity with PII, a gene previously described from A. oligospora, and we conclude that this gene is likely a PII ortholog.
一种中性丝氨酸蛋白酶（命名为 Aoz1）从云南省土壤中分离得到的少孢节丛孢菌（Arthrobotrys oligospora）菌株中被纯化为均质。纯化的蛋白质显示约 38000 道尔顿的分子量、pI 4.9，并在 45°C 和 pH 6-8 时显示最佳活性。该蛋白可水解明胶、酪蛋白和显色底物偶氮胶原，并可在体外固定线虫（潘格尔勒斯雷迪维乌斯 L. [古迪]）。在培养基中发现活性水平随明胶浓度的增加而增加。扫描电子显微镜显示，用纯化的蛋白酶处理的线虫表皮发生了剧烈的结构变化。通过 N 端序列分析获得的部分肽序列用于设计用于分离编码成熟蛋白酶的 cDNA 基因的简并引物。对 cDNA 和相应基因组序列的分析表明，该基因与先前从少孢节丛孢菌描述的 PII 具有 97%的同一性，我们得出结论，该基因可能是 PII 的同源物。
