|
Type |
Poster Presentation |
Area |
생명화학 |
Room No. |
포스터발표장 |
Time |
4월 21일 (금요일) 13:00~14:30 |
Code |
BIO.P-305 |
Subject |
Biochemical Characterization of a hormone sensitive lipase homolog (Lg36) from Lactococcus garvieae |
Authors |
Wang Ying, 김두헌1,* 숙명여자대학교 화학과, Korea 1숙명여자대학교 이과대학 화학과, Korea |
Abstract |
A novel oligomeric carboxylesterase (Lg36) from Lactococcus garvieae was identified, over-expressed in Escherichia coli, characterized, and immobilized for industrial application. Primary sequence analysis revealed that Lg36 has a catalytic triad consisting of Ser159, Asp256 and His286. Phylogenetic analysis showed that Lg36 was clustered in a branch of the family IV group. Biochemical properties of Lg36 were investigated by performing mass spectrometry, circular dichroism (CD), fluorescence, gel filtration, and pH indicator-based colorimetric assays. Furthermore, four important residues (F207, L208, F216, L258) involved in the formation of substrate-binding pocket were identified from structure analysis and mutagenesis. Cross-linked enzyme aggregates (CLEAs) of Lg36 were prepared by precipitating the enzyme with ammonium sulfate and subsequent cross-linking with glutaraldehyde. In addition, hybrid magnetic cross-linked enzyme aggregates (HM-CLEAs) of Lg36 were developed by co-aggregation of enzyme aggregates with magnetite nanoparticles and subsequent chemical cross-linking with glutaraldehyde. Interestingly, HM-CLEAs of Lg36 was enhanced in stability and recycling compared to CLEAs, which could be used for industrial application. |
E-mail |
yingaabb@yahoo.com |
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