| Title | Exploration and functional expression of homologous lipases of Candida antarctica lipase B | ||
| Author | Seongsoon Park* | ||
| Address | Department of Chemistry, Center for NanoBio Applied Technology, and Institute of Basic Sciences, Sungshin Women’s University, Seoul 01133, Republic of Korea | ||
| Bibliography | Korean Journal of Microbiology, 51(3),187-193, 2015 | ||
| DOI | http://dx.doi.org/10.7845/kjm.2015.5037 | ||
| Key Words | heterologous expression, homology, lipase | ||
| Abstract | Candida (also known as Pseudozyma) antarctica lipase B (CAL-B) has been intensely studied in academic and industrial fields. However, the research related to its homologous enzymes has been rarely reported. In the current investigation, protein sequence similarity search of CAL-B has been conducted and six homologous protein sequences were identified. After the syntheses of their codon-optimized genes, the synthetic genes have been cloned into a periplasmic expression vector to express in Escherichia coli. Among six homologous sequences, four sequences were successfully expressed in E. coli. The hydrolytic activities of the expressed proteins towards 4-nitrophenyl acetate and 4-nitrophenyl butyrate were measured and compared with those of CAL-B to identify whether the expressed proteins work as a hydrolase. It has been revealed that the expressed proteins can hydrolyze the substrates and the specific activities were determined as (1.3– 30) × 10-2 μmol/min/mg, which are lower than those of CAL-B. Among these homologous enzymes, Pseudozyma hubeiensis SY62 exhibits the comparable enantioselectivity to that of CAL-B towards the hydrolysis of (±)-1-phenylethyl acetate. | ||
| Download PDF | 51(3)_01_p.187-193.pdf | ||