Successful production of recombinant buckwheat cysteine-rich aspartic protease in Escherichia coli | Zendy

Mira Milisavljević | Zendy; Dražen Papić | Zendy; Gordana Timotijević | Zendy; Vesna Maksimović | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

Successful production of recombinant buckwheat cysteine-rich aspartic protease in Escherichia coli

Author(s) -

Mira Milisavljević,

Dražen Papić,

Gordana Timotijević,

Vesna Maksimović

Publication year - 2009

Publication title -

journal of the serbian chemical society

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.227

H-Index - 45

eISSN - 1820-7421

pISSN - 0352-5139

DOI - 10.2298/jsc0906607m

Subject(s) - inclusion bodies , recombinant dna , cysteine , escherichia coli , protease , biochemistry , chemistry , fusion protein , cysteine protease , enzyme , strain (injury) , biology , anatomy , gene

Herein, the expression of recombinant cysteine-rich atypical buck- wheat (Fagopyrum esculentum) aspartic protease (FeAPL1) in five Escherichia coli strains differing in their expression capabilities is presented. It was shown that the expression success depended highly on the choice of FeAPL1 fusion partner. His6-FeAPL1 was produced in large quantities as an insoluble protein localized in inclusion bodies. On the other hand, MBP-FeAPL1 was localized in both the cytoplasm and inclusion bodies in BL21 and Rosetta-gami strains. Only purified soluble MBP-FeAPL1 from Rosetta-gami cells showed proteo- lytic activity at pH 3.0 with BSA as the substrate. The results also indicated that FeAPL1 contained a PRO segment that had to be removed for the enzyme activity to appear. The activity of FeAPL1 produced in the Rosetta-gami strain, which enables disulfide bond formation, indicated the importance of the twelve cysteine residues for correct folding and functionality.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research