香蕉果皮中过氧化物酶的提取与纯化研究

On the extraction and purification ofperoxidase from the banana peel

为使香蕉果皮废物再利用,通过表面活性剂沉淀技术对香蕉果皮中的过氧化物酶进行提取,并对提取液和粗酶液的蛋白质含量以及酶活性进行测定。结果显示司盘80(span-80)提取过氧化物酶的最佳回收溶剂为丙酮、p H值为6.0、离子强度为2 mmol/L、中和剂浓度为0.15 mmol/L。该方法为果皮废物利用提供了理论指导。

The given paper is to engage itself in a study of the advantage of the banana peel waste to extract the peroxidase from the said peel by way of surfactant precipitation,which just demands low surfactant concentration rate and simple operation.To fulfill the above said operation,there is only one need,that is,to add the non-ionic surfactant span-80 to the crude enzyme liquid taken from the banana peel to form the protein-surfactant complex,and extract and purify the peroxidase in the crude enzyme solution,so as to analyze the enzyme activity and its content by calculating the enzyme activity recovery rate and the protein recovery rate with the recovery absolvent.At the same time,we have also managed to change the span-80 mass concentration rate via the single factor method,so as to recover the solvent,the pH value,the ionic strength,the neutralizer concentration rate,as well as to explore the impact of the above said factors on the efficiency of the span-80 extraction and peroxidase purification.The results of our study indicate that the increase in the surfactant concentration is beneficial for its combination with enzyme.And,thereby,it is also favorable for improving the protein recovery and enzyme activity recovery,though the pH value may have a significant effect on the peroxidase purification with a certain amount of KCl turning to promote the formation of complexes,whereas the neutralizing agents can be used to neutralize the charge in the extraction.As the concentration rate of the neutralizing agent sodium chloride tends to increase from 0.05 mol/L to 0.3 mol/L,the recovery rate of the protein and enzyme activity may tend first to increase and then decrease,signifying that too high or too low concentration rate of neutralizing agent is not conducive to the extraction and purification of the enzyme.Therefore,the results of our study may show that the optimum recovery solvent,the pH value,the ionic strength,the surfactant mass concentration and neutralizer concentration rate of span-80 to extract peroxidase should be made to promote the extraction and purification,including acetone,the pH value of 6.0,2 mmol/L,14 mg/L and 0.15 mmol/L,respectively.Hence,the given method can provide a proper theoretical guidance for the utilization of such peel waste resources.