Secretory expression of a heterologous nattokinase in Lactococcus lactis.
Nattokinase has been reported as an oral health product for the prevention of atherosclerosis. We developed a novel strategy to express a nattokinase from Bacillus subtilis in a live delivery vehicle, Lactococcus lactis. Promoter P( nisZ) and signal peptide SP(Usp) were used for inducible and secretory expression of nattokinase in L. lactis. Western blotting analysis demonstrated that nattokinase was successfully expressed, and about 94% of the enzyme was secreted to the culture. The recombinant nattokinase showed potent fibrinolytic activity, equivalent to 41.7 urokinase units per milliliter culture. Expression and delivery of such a fibrinolytic enzyme in the food-grade vehicle L. lactis would facilitate the widespread application of nattokinase in the control and prevention of thrombosis diseases.
Liang
Xiaobo
X
Center for Metabolic Engineering of Microorganisms, Institute of Microbiology, Chinese Academy of Sciences, Beijing, PR China.
Zhang
Lixin
L
Zhong
Jin
J
Huan
Liandong
L
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001709:Biotechnology / Q000379:methods; D005343:Fibrinolytic Agents / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D013294:Lactococcus lactis / Q000201:enzymology* / Q000235:genetics; D009561:Nisin / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013381:Subtilisins / Q000235:genetics* / Q000378:metabolism*; D013927:Thrombosis / Q000188:drug therapy
D001426:Bacterial Proteins; D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; D009561:Nisin; D013381:Subtilisins; C053771:nattokinase