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Parametric optimization on the sensitive determination of midgut α-amylase in larvae of Mythimna separata Walker
Author(s): 
Pages: 981-988
Year: Issue:  10
Journal: ACTA ENTOMOLOGICA SINICA

Keyword:  粘虫α-淀粉酶参数优化动力学常数;
Abstract: 针对粘虫Mythimna separata中肠α-淀粉酶筛选了11种不同参数组合的3,5-二硝基水杨酸活性测定方法,并对其中最适组合的各个参数进行了优化.结果表明:在离体测定条件下,粘虫中肠α-淀粉酶活性的最优化测定参数为0.03 mol/L磷酸盐缓冲液(pH 8.0,含有55 mmol/L NaCl)、温度45℃、吸收波长480 nm.Ca2+对α-淀粉酶活性具有抑制作用.该优化法能够显著降低粘虫、德国小蠊Blattella germanica、黄粉虫Tenebrio molitor、淡色库蚊Culex pipiens pallens和家蝇Musca domestica等昆虫α-淀粉酶的米氏常数Km值,且粘虫和德国小蠊α-淀粉酶的Vmax值增大,但黄粉虫、淡色库蚊和家蝇α-淀粉酶的Vmax值均明显减小.结果说明,在该优化体系下,粘虫α-淀粉酶与底物的亲和力增强,最大反应速度增大,测定酶活性的准确性和灵敏度显著提高;同时该优化体系也可作为测定德国小蠊α-淀粉酶活性的优化方法,但不适合作为黄粉虫、淡色库蚊和家蝇α-淀粉酶的最优化测定方法.
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