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接骨草蔗糖酶同工酶的分离纯化及其部分性质研究 被引量:4

Isolation, Purification and Some Properties of the Isozymes of Invertase in Sambucus chinensis Lindl
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摘要 从接骨草幼叶中提取的蔗糖酶粗酶液经硫酸铵分级沉淀、DEAE-Sepharose离子交换层析、SephacrylS-200 凝胶过滤层析纯化,得到3种同工酶.对3种同工酶进行SDS-PAGE和IEF电泳均显示一条蛋白带,其亚基分子量分别为45.8 KD,44.3 KD,43.2 KD;等电点分别为pH 4.0,pH 3.8,pH 3.75.凝胶过滤测得3种同工酶全酶分子量分别为91.2 KD,89.1 KD,87.4 KD,表明3种同工酶均由2个相同亚基组成,其表观Km分别为30 mmol/L, 57.1 mmol/L,65 mmol/L;Vmax分别为98μg/min,48 μg/min,35μg/min.同工酶Ⅰ和同工酶Ⅲ在pH4.4-5.0 时有最大活性,同工酶Ⅱ则在pH3.9-4.2时有最大活性.3种同工酶均在pH3-6范围内比较稳定.同工酶Ⅰ和同工酶Ⅱ在48-52℃有最大活性,同工酶Ⅲ在50-55℃有最大活性.3种同工酶均在50℃以下有较好的稳定性. Invertase solution extracted from the leaves of Sambucus chinensis Lindl has been isolated and purified by ammonium sulphate precipitation, DEAE-Sepharose column chromatography, and SephacrylS200 gel filtration, and three isozyme of invertase were obtained. The purified isozymes were homogeneous as shown in SDS-PAGE and IEF electrophoresis. The molecular weight of the isozymes determined by SDS-PAGE were 45.7 KD, 44. 9 KD, 44. 1 KD and gel filtration were 91.2 KD, 89.1 KD, 88. 1 KD respectively. The PI determined by IEFelectrophoresis were pH4.0, pH3.8, pH3.75 respectively. The Km of the isozymes were 30 mmol/L, 57.1 mmol/L, 65 mmol/L and Vmax were 98 μg/min, 48 μg/min, 35 μg/min respectively. The optimum pH range of isozyme Ⅰ and isozymeⅢ was pH4. 4 - pH5. 0, and isozymeⅡ was pH3. 9 - pH4. 2. The optimum temperature range of isozymeIand isozymeⅡwas 48 ℃- 52 ℃, and isozymeⅢ was 50℃- 55℃. Three isozymes were stable at the range of pH3-pH6 and under 50℃.
出处 《西南师范大学学报(自然科学版)》 CAS CSCD 北大核心 2005年第6期1116-1121,共6页 Journal of Southwest China Normal University(Natural Science Edition)
基金 重庆市科委资助项目CSTC 2004AC1012
关键词 接骨草叶片 蔗糖酶 同工酶 分离纯化 性质 leaves of Sambucus chinensis Lindl invertase isozyme isolation and purification properties
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参考文献10

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二级参考文献5

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