A comparison between the recombinant expression and chemical synthesis of a short cysteine-rich insecticidal spider peptide
AUTOR(ES)
Clement, Herlinda, Flores, Vianey, Diego-Garcia, Elia, Corrales-Garcia, Ligia, Villegas, Elba, Corzo, Gerardo
FONTE
J. Venom. Anim. Toxins incl. Trop. Dis
DATA DE PUBLICAÇÃO
11/08/2015
RESUMO
The choice between heterologous expression versus chemical synthesis for synthesizing short cysteine-rich insecticidal peptides from arthropods may impact the obtainment of yields and well-folded bioactive molecules for scientific research. Therefore, two recombinant expression systems were compared to that of chemical synthesis for producing Ba1, a cysteine-rich spider neurotoxin.
The transcription of the insecticidal neurotoxin Ba1 was obtained from a cDNA library of venom glands of the spider
Both expression systems pQE40 and pET28a+ expressed the His-tagged recombinant protein products, HisrDFHRBa1 and HisrBa1, respectively, as inclusion bodies. The recombinant proteins HisrDFHRBa1 and HisrBa1 presented respective molecular masses of 28,289 and 8274.6 Da, and were not biologically active. These results suggested that both HisrDFHRBa1 and HisrBa1 were oxidized after cell extraction, and that their insecticidal activities were affected by their N-terminal pro-peptides and different disulfide bridge arrangements. The respective protein expression yields for HisrDFHRBa1 and HisrBa1 were 100 μg/L and 900 μg/L of culture medium. HisrBa1 was reduced and folded under
The two recombinant insecticidal peptides and the one synthesized chemically were as active as the native Ba1; however, toxin yields differed drastically.
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