Journal of Proteomics & Bioinformatics
Open Access
ISSN: 0974-276X
ISSN: 0974-276X
Phuong Le Trinh, Quynh Diep Dinh and Yen Bao Pham
VNU University of Science, Vietnam
Posters & Accepted Abstracts: J Proteomics Bioinform
Lipase enzyme, secreted by Helicobacter pylori, is reported to play an important role in the bacterial survival and the pathogenic�s ulcer developing-activities of the bacteria. Hence, it is considered a molecular target for H. pylori inhibitor screening, which consequently requires large amount of active lipase. In this study, we designed primers to clone H. pylori lipase gene in four different constructs containing His-tag, signal peptide sequence or TEV cleavage sequence based on the vector pET22b+ and afterward examined various strategies to express the recombinant lipase in heterologous system E.coli BL21 (DE3) RIL. The results indicated that additives (i.e IPTG; Glucose, Glycylglycine) and induction method (i.e heat shock, high cell density expression) had different effects on four constructs, of which combination of glucose and high cell density expression greatly improved total yield in nearly all constructs. In the most responsive construct, recombinant lipase was comprised of 50% total protein, 10% of which was in the soluble fraction. The Ni-Sepharose column purification was successfully used to collect pure lipase (15 mg/L cell culture) from the soluble fraction.
Phuong Le Trinh gained her Bachelor’s degree from VNU University of Science, Hanoi, and then completed her Master’s from the University of Bonn, Germany. She is now an Early Researcher at The Key Laboratory of Enzyme and Protein, VNU University of Science, Hanoi. She has been co-authoring in nearly 10 papers in Vietnamese journals and in 2 papers in ISI journals and presenting at two international conferences.
Email: violetph2009@gmail.com