光谱学与光谱分析, 2017, 37 (6): 1896, 网络出版: 2017-07-10  

微波场中卵清蛋白糖基化产物不均匀性的光谱分析

Inhomogeneity of Ovalbumin Glycosylation Products in Microwave Field with Spectral Analysis
作者单位
1 南昌大学食品科学与技术国家重点实验室, 江西 南昌 330047
2 江西师范大学生命科学学院, 江西 南昌 330022
摘要
以卵清蛋白与葡萄糖混合体系为研究对象, 构建微波场中二者糖基化反应模型, 采用红外、 荧光、 紫外等方法研究微波场中卵清蛋白糖基化产物不均匀性。 将卵清蛋白与葡萄糖按照1∶1的比例混合成5%的溶液, 经过48 h冷冻干燥形成5 mm厚的糖基化干样模型, 在微波炉中反应, 按五个反应区将样品一分为五进行实验分析, 1号区域为微波炉中心位置, 2号区域靠近磁控管, 3, 4和5号区域在远离磁控管一端。 结果表明, 蛋白与糖混合物在560 W微波下9 min时发生不均匀的糖基化反应; SDS-PAGE电泳图和自由氨基含量表明微波作用下的糖基化确实存在较大不均匀性; 通过荧光和三维荧光光谱可知糖基化反应产生了新的荧光性物质, 其中3, 4和5号位置的样品产生了新的较大的荧光峰, 而荧光性物质是进行到糖基化高级阶段的反应产物, 说明这些样品产生了高级糖基化产物; 红外光谱分析表明有新的N—H键生成; 紫外分析表明在糖基化反应模型平面中, 3号位置的反应程度最高。
Abstract
In this experiment, the mixed system of ovalbumin and glucose were constructed as the research object of glycosylation reaction model. Then the infrared, fluorescence and UV were used to analyze the product of different position in microwave field. Ovalbumin and glucose were mixed into 5% solution with the proportion of 1∶1. Then it formed into a plane model with a thickness of 5mm after freeze-drying for 48h. The reaction was taken in the microwave oven, the reacted samples were divided into five regions, including No.1 in the center of the microwave oven, No. 2 near the magnetron, No. 3, No.4 and No.5, away from the magnetron. The results showed that the mixture of protein and sugar reacted unevenly under the condition of 560 W for 9 minutes in microwave. SDS-PAGE electrophoresis diagram and the content of free amino showed that with the action of microwave glycosylation does exist larger nonuniformity; by analysing fluorescence and three-dimensional fluorescence spectrum, it was found that glycosylation reaction created new fluorescent substances, including 3, 4, and 5 location created new larger fluorescence peaks which were from the advanced products of the glycosylation; infrared spectrum analysis showed that new N—H key was generated; UV analysis showed that in plane of glycosylation reaction model, location 3 was the highest level of response.

钟比真, 涂宗财, 王辉. 微波场中卵清蛋白糖基化产物不均匀性的光谱分析[J]. 光谱学与光谱分析, 2017, 37(6): 1896. ZHONG Bi-zhen, TU Zong-cai, WANG Hui. Inhomogeneity of Ovalbumin Glycosylation Products in Microwave Field with Spectral Analysis[J]. Spectroscopy and Spectral Analysis, 2017, 37(6): 1896.

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