[Optimization of prokaryotic expression conditions of Leptospira interrogans trigeminy genus-specific protein antigen based on surface response analysis].
Lipoproteins LipL32 and LipL21 and transmembrane protein OMPL1 have been confirmed as the superficial genus-specific antigens of Leptospira interrogans, which can be used as antigens for developing a universal genetic engineering vaccine.


OBJECTIVE
In order to obtain high expression of an artificial fusion gene lipL32/1-lipL21-ompL1/2, we optimized prokaryotic expression conditions.


METHODS
We used surface response analysis based on the central composite design to optimize culture conditions of a new antigen protein by recombinant Escherichia coli DE3.The culture conditions included initial pH, induction start time, post-induction time, Isopropyl beta-D-thiogalactopyranoside (IPTG) concentration, and temperature.


RESULTS
The maximal production of antigen protein was 37.78 mg/l. The optimal culture conditions for high recombinant fusion protein was determined: initial pH 7.9, induction start time 2.5 h, a post-induction time of 5.38 h, 0.20 mM IPTG, and a post-induction temperature of 31 degrees C.


CONCLUSIONS
Surface response analysis based on CCD increased the target production. This statistical method reduced the number of experiments required for optimization and enabled rapid identification and integration of the key culture condition parameters for optimizing recombinant protein expression.
National Key Laboratory of Infectious Disease Diagnosis and Therapy, the First Affiliated Hospital of Zhejiang University, Hangzhou 310009, China.
Jiang
J
Wang
Dongjiao
D
Luo
Aihua
A
Sun
Jie
J
Yan
D000954:Antigens, Surface / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D007921:Leptospira interrogans; D016014:Linear Models; D008954:Models, Biological; D011387:Prokaryotic Cells / Q000378:metabolism*; D013045:Species Specificity; D013696:Temperature; D013997:Time Factors
D000954:Antigens, Surface; D007544:Isopropyl Thiogalactoside