Construction and Activity Screening of Lipase Mutagenesis Library from Candida rugosa

• Main Author: 楊舒涵
• Other Authors: Shye-Jye Tang
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/66732519949268964501

碩士 === 國立臺灣海洋大學 === 生物科技研究所 === 100 === The yeast Candida rugosa produces five extracellular lipases (LIP1-LIP5), showing diverse enzyme activity but high gene similarity. The previous researcher recombine LIP F and LIP4, producing 13 chimera lipases: NL-A~NL-M, to study the relationship between sequence and structure-activity. There are only NL-B, NL-J and NL-L have activity. To understand the substrate specificity and activity between these new lipase, the previous researcher recombinant the NL-B and NL-L trying to higher the activity of new lipase but failed. There are only 20 amino aicds different between NL-BL and NL-L and the difference is on the lid structure. To understand the different between NL-BL and NL-L, use the Quick Change Mutagenesis PCR to mutant the different sequence from NL-BL to NL-L. The result shows that the activity of mutant are successfully enhance 2-5 folds.