【作者】 刘朋虎；

【导师】 梁勤；

【作者基本信息】 福建农林大学 ， 特种经济动物饲养， 2007， 硕士

【摘要】 蜜蜂白垩病（bee chalkbrood disease）是由蜜蜂球囊菌（Ascosphaera apis Olive & Spiitoir）引起的蜜蜂幼虫的真菌传染病。目前为止，还没有治疗该病的有效方法。在无法控制的条件下，该病仍呈自然流行状态。几丁质酶在昆虫病原真菌的入侵中有重要作用。我们从10株蜜蜂球囊菌中筛选出一株高产几丁质酶的菌株，并且对其几丁质酶进行了诱导、分离、纯化和酶学性质的研究。经过硫酸铵盐析、SephadexG-75凝胶层析、DE-52离子交换层析得到纯化倍数为6.7的几丁质酶。该几丁质酶的最适反应温度为40℃，在35℃～45℃之间能保持40％以上的活力，但是这种几丁质酶对高温比较敏感；在pH7.5时，蜜蜂球囊菌几丁质酶的活性最高，在pH4.5以下几乎检测不到活力，在pH9.0以上酶活力在20％以下，在pH6～9范围内几丁质酶活力都在40％以上；在0～100mmol／L范围内，一价金属离子Na⁺、K⁺对几丁质酶活力影响均表现为先促进后抑制；在浓度为1～10mmol／L范围内，二价金属离子Mg²⁺、Mn²⁺对其活力起抑制作用，Zn²⁺对几丁质酶活力基本没有影响，Ca²⁺、Fe²⁺对几丁质酶活力起促进作用；在浓度1～10mmol／L范围内，三价金属离子Al³⁺对蜜蜂球囊菌几丁质酶活力起促进作用，Fe³⁺对几丁质酶活力基本没有影响。更多还原

【Abstract】 Bee chalkbrood disease was a fungal epidemic disease of apian larva which was caused by Ascosphaera apis Olive&Spiltoir. Up to now , there were no effective methods to cure this disease. Under this conditions, this disease was still natural popular. Chitinase has a important role when pathogenetic fungi infected insects. One strain was selected from a total of ten stains Ascosphaera apis Olive & Spiltoir that could produce chitinase. The chitinase was purified , and its Properities was studied. After salt out of （NH4）₂SO₄, ion-exchange chromat-ography on DE-52 and Gel-filtration chromatography on Sephadex G-75, the fold was 6. 7. The chitinase exhibited optimum catalytic activity 40℃. It was retained above 40% activity between 35℃～45℃, but it was sensitive to high temperature; Its optimal pH was 7. 5, its activity could not be detected when pH below 4. 5 and above pH9. 0 its activity below 20%, and its activity was above 40% when pH between pH 6～9. When concentration between 1～100mmol/L, metal metal ions Na⁺、K⁺ enhanced the enzyme activity at first and then inhibited the enzyme activity. When concentration between 1～10mmol/L, Ca²⁺、Fe²⁺、Al³⁺ enhanced the enzyme activity, Mg²⁺、Mn²⁺ inhibited the enzyme activity, Zn²⁺、Fe³⁺ has no effect to the enzyme activity.更多还原