Effects of Cecropin Peptides on Aquatic Animal Pathogens and Shrimp Hemocytes.

Ho, Shih-Hu; Lin, Chia-Zong; Chen, Yu‐Chih; Song, Yen-Ling

doi:10.3147/jsfp.37.7

Cited by 2 publications

(1 citation statement)

References 32 publications

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“…Reports of antimicrobial activity against E. coli range from 0.5-0.9 μM for cecropin AD (Chen et al 2009) and cecropin A (Silvestro et al 2000). Activity for cecropin A against Vibrio ordalii and Vibrio alginolyticus was 2.06 μM Schmitt et al 2008) and 0.98-7.81 μM for cecropin against Vibrio (Ho et al 2002). The choice of pTYB11 for cloning and expression allowed the intein fusion to the N-terminus of cecropin.…”

Section: Resultsmentioning

confidence: 98%

Intein-mediated expression of cecropin in Escherichia coli

Díaz¹,

Venturini²,

Marchetti³

et al. 2012

Electro Journal of Biotech

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Different strategies have been used to overcome the difficulties to produce antimicrobial peptides. Here we used Intein Mediated Purification with an Affinity Chitin-binding Tag (IMPACTSystem, New England Biolabs) for the expression of the antimicrobial peptide cecropin to reduce its sensitivity to intracellular proteases and use its inducible self-cleaving capability to remove the carrier. Cecropin was cloned into suitable expression vector pTYB11, and expression induced by IPTG in Escherichia coli ER2566. The use of 22ºC induction allowed the expression of cecropin with its intein carrier in soluble form. Cell extracts were purified by chitin affinity chromatography and intein-mediated splicing of the target protein was achieved by thiol addition, obtaining a final yield of 2.5 mg cecropin/l. Cecropin cleaved from the intein had its proper biologically active form, showing a micromolar antimicrobial activity against Vibrio ordalii, Vibrio alginolyticus and Escherichia coli.

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