de Freitas Soares Filippe Elias, de Queiroz José Humberto, Braga Fabio Ribeiro, Dos Santos Lima Walter, Zamprogno Tatiana Tonini, de Araújo Jackson Victor
Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, MG, Brasil.
BMC Res Notes. 2014 Nov 18;7:811. doi: 10.1186/1756-0500-7-811.
The predatory nematophagous fungus Arthrobotrys sinensis (SF53) produces three proteases with nematicidal activity when grown on solid media culture. However, the proteolytic profile produced by this fungus, when grown in liquid culture medium remains unknown.
Thus, the objective of this work was to evaluate the production of proteases from nematophagous fungus Arthrobotrys sinensis in liquid medium and its nematicidal activity on first stage larvae of A. vasorum. Proteases were obtained in its crude form, using Whatman no.1 filter paper, followed by centrifugation for 5 min at 10 × g and 4°C. A zymogram was performed with co-polymerized casein in an acrylamide gel as substrate. An in vitro assay to evaluate the nematicidal action of the proteases of A. sinensis (SF53) produced in liquid medium on A. vasorum L1 was conducted. By the analysis of the zymogram, it was observed a single halo at the beginning of digestion of the gel, suggesting that the three proteases of SF53 are produced in an enzymatic complex of large molecular weight. Regarding nematicidal activity, within 24 hours, the proteases produced in liquid medium of A. sinensis (SF53) showed a percentage reduction of 64% on the number of L1 of A. vasorum.
In the present work, it is suggested that the three proteases of SF53 are produced in an enzymatic complex and was also demonstrated that these enzymes were effective in destroying A. vasorum L1.
捕食性线虫真菌中华节丛孢（SF53）在固体培养基上生长时会产生三种具有杀线虫活性的蛋白酶。然而，该真菌在液体培养基中生长时产生的蛋白水解谱仍然未知。
因此，本研究的目的是评估线虫真菌中华节丛孢在液体培养基中蛋白酶的产生及其对血管圆线虫一期幼虫的杀线虫活性。使用Whatman 1号滤纸以粗提形式获得蛋白酶，然后在10×g和4°C下离心5分钟。以丙烯酰胺凝胶中共聚的酪蛋白为底物进行酶谱分析。进行了一项体外试验，以评估在液体培养基中产生的中华节丛孢（SF53）蛋白酶对血管圆线虫L1的杀线虫作用。通过酶谱分析，在凝胶消化开始时观察到一个单一的晕圈，表明SF53的三种蛋白酶是在大分子量的酶复合物中产生的。关于杀线虫活性，在24小时内，中华节丛孢（SF53）液体培养基中产生的蛋白酶使血管圆线虫L1的数量减少了64%。
在本研究中，表明SF53的三种蛋白酶是在酶复合物中产生的，并且还证明这些酶对破坏血管圆线虫L1有效。
