119th General Meeting of the KCS

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