| Summary: | 碩士 === 國立中興大學 === 化學工程學系所 === 102 === It is said that enzyme’s His-tag will absorb certain proteins while immobilization, and save the cost of purification at the same time. In this study, iminodiacetic acid (IDA) is used to modify Immobead 150 as chelating agent to cover the substrate of the epoxy group, and it is concluded that when a protein concentration is between 2.5-3mg/ml, enzyme could be better immobilization.
Two enzymes Protein switch RG13 and L-N-carbamoylase are used for the following two reasons. First, Protein switch RG13 could specify the concentration of maltose, and induce the activity of different enzymes. Second, L-N-carbamoylase could catalyze a variety of primary N-carbamoyl-L -amino acid, and it can be used to catalyze the production of L-HPA. It is known that the L-HPA can be used as precursor of antihypertensive drug, which has high economic value nowadays.
Under the same condition, compared the selectivity adsorption of the two enzymes, the selectivity adsorption of L-NCA is 1.647, and RG13 is 1.07, which is 53.9% more than that of RG13. Therefore, L-NCA is apparently better than RG13. Moreover, in L-NCA immobilization, it took about 2 to 3 hours to form physical adsorption by means of IMAC, and achieved the maximum amount of adsorption after 4 or 5 hours.
To summary, the results show that the time of adsorption is related with the enzyme activity, and by using 0.3M iminodiacetic acid(IDA) to modify Immobead 150 could achieve the maximum activity after five hours of adsorption.
|
|---|
