黄蓝状菌几丁质酶E2纯化、性质及抗菌活性研究

黄蓝状菌在SMCS液体培养基中置于恒温摇床（200r／min，27．5C）上培养13天，诱导产生大量的几丁质酶。培养滤液经硫酸铵分级沉淀，DEAE--Sepharose FastFlow阴离子交换层析、Phenyl—Sepharose Fast Flow疏水层析、Sephacryl S-100分子筛层析得到了凝胶电泳（SDS—PAGE）谱带单一的几丁质酶E2，其分子量为32KD。其最适反应温度为50℃，最适pH值是5。Mn^2＋对其有明显的激活作用，Cu^2＋、FeCu^2＋对其有强烈的抑制作用。抗菌活性显示．其对供试病原菌具有明显的抑菌作用。

Purification,properties and antifungal activity of one chitinase produeced by Talaromyces flavus

One extracellular chitinase E, was purified to SDS--PAGE homogeneous by ammonium sulfate fraction, DEAE Sepharose Fast Flow Chromatography, Phenyl-Sepharose Fast Flow Chromatography and Sephacryl S- 100 charomatogra phy when Talaromyces fiavus was grown in SMCS broth at 27.5℃ on a rotary shaker at 200r/min for 13 days. The molecular weight of the chitinase was about 32kD. The property and antifungal activity of chitinase were tested in tbis study,the results were as the followings：The optimal pH and temperature of the enzyme for hydrolysis of chitin are 5 and at 50℃ respectively. Different metal ions showed different effects on the chitinase activities, Mn2＋ evidently increased activity of the chitinases, Na＋,Ca2＋ and K＋ increased partially activitiy of the chitinase , whereasCu2-, FeCu2＋ caused inhibition heavily. Thechitinases appeared to be antifungal activity against tested fungi and more effective against fungi of Fusarium oxysporum Schlecht, Botryosphaeria berengeriana f. sp. Piricola Alternaria logipes and Verticillium dahliae.