Characterization of lysine-tagged Bacillus stearothermophilus leucine aminopeptidase II immobilized onto carboxylated gold nanoparticles |
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| Authors: | Chien-Li Wu Yi-Pu Chen Jia-Ci Yang Huei-Fen Lo Long-Liu Lin |
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| Affiliation: | aDepartment of Applied Chemistry, National Chiayi University, 300 University Road, Chiayi 60083, Taiwan;bDepartment of Food and Nutrition, Hungkuang University, Taichung 433, Taiwan |
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| Abstract: | Bacillus stearothermophilus leucine aminopeptidase II tagged C-terminally with either tri- or nona-lysine (BsLAPII-Lys3/9) was constructed and over-expressed in Escherichia coli M15 (pRep4). The recombinant enzymes were purified to homogeneity by nickel-chelate chromatography and their molecular masses were determined to be approximately 45 kDa by SDS/PAGE. Surface modification of colloidal gold with 16-mercaptohexadecanoic acid was employed to generate the carboxylated nanoparticles. BsLAPII-Lys9 was efficiently immobilized onto the carboxylated gold nanoparticles (AuNP-COOH) and the obtained bioconjugate showed excellent biocatalytic activity in the immobilized form. Additionally, the bioconjugate material exhibited a significant enhancement in temperature stability and could be reused over 5 successive cycles. |
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| Keywords: | Bacillus stearothermophilus Leucine aminopeptidase Escherichia coli Lysine tag Gold nanoparticles Immobilization |
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