Enhanced Expression of Lipoxygenase in Escherichia coli by Co-Expressing Antioxidases

doi:10.3969/j.issn.1673-1689.2019.10.004

Home > Archive>Volume 38, Issue 10, 2019 >23-29. DOI:10.3969/j.issn.1673-1689.2019.10.004

Enhanced Expression of Lipoxygenase in Escherichia coli by Co-Expressing Antioxidases
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                        10.3969/j.issn.1673-1689.2019.10.004
                    
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Based on the potential hazards of lipoxygenase（LOX） activity to host strain，three antioxidases（two superoxide dismutases and a catalase） are respectively co-expressed to improve the expression of lipoxygenase expression in Escherichia coli Rosetta（DE3）. The superoxide dismutase genes（sodB and sodM） from P. aeruginosa BBE and catalase gene（katE） from E. coli are cloned into expression plasmid pRSFDuet-1，yielding the plasmids pRSF-sodB，pRSF-sodM and pRSF-katE. The recombinant plasmids are transformed into N6（a LOX expressing strain generated by E. coli） to obtain strains N6-B，N6-M and N6-K，respectively. Under the induction with 1 mmol/mL ITPG at 20 ℃，the total LOX activities of N6-B，N6-M and N6-K reach 21.6，28.1 and 7.1 U/mL respectively. The yield of LOX in N6-B and N6-M is increased by 83% and 138% in contrast to N6（11.8 U/mL），respectively. The optimized induction by using orthogonal test is as follow：OD₆₀₀=2.5，IPTG=2 mmol/mL，and inducing temperature is 20 ℃. The results show that the co-expression of superoxide dismutases could effectively improve the expression of LOX in E. coli，which provides a new idea for efficient heterologous expression of this enzyme.

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QIU Fangfang, LIU Song, DU Guocheng, CHEN Jian. Enhanced Expression of Lipoxygenase in Escherichia coli by Co-Expressing Antioxidases[J]. Journal of Food Science and Biotechnology,2019,38(10):23-29.

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