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GAP-initiated constitutive expression of a novel plectasin-derived peptide MP1106 by Pichia pastoris and its activity against Streptococcus suis

Paper ID Volume ID Publish Year Pages File Format Full-Text
34398 45023 2015 9 PDF Available
Title
GAP-initiated constitutive expression of a novel plectasin-derived peptide MP1106 by Pichia pastoris and its activity against Streptococcus suis
Abstract

•Constitutive expression of rMP1106 by Pichia pastoris.•Higher production rate of rMP1106 from the GAP promoter.•Lower costs, risks and toxicity than AOX promoter depending on methanol induction during expression.•Stronger antimicrobial activity of rMP1106 against Streptococcus suis than plectasin or antibiotics.•High stability of rMP1106 in a broad range of pH, temperature and ionic strength.

MP1106 is a novel variant of plectasin with potent activity against some Gram-positive pathogenic bacteria, especially for Streptococcus suis and Staphylococcus aureus. MP1106 was successfully expressed in Pichia pastoris X-33 using the glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter. The total protein concentration and antimicrobial activity were 998.3 mg/L and 51,200 AU/mL, respectively. After purification by cation exchange chromatography, recombinant MP1106 (rMP1106) with a concentration of 100.13 mg/L and a purity of 95.2% was obtained from fermentation culture. Its antimicrobial spectrum was similar to that of plectasin, and the antibacterial activity against S. suis CVCC3309 was enhanced up to 29-fold over that of plectasin. In addition, 97.9% of the S. suis CVCC606 bacteria were killed by 16× minimal inhibitory concentration (MIC) rMP1106 within 6 h. The percentage hemolysis by rMP1106 against mouse erythrocytes was 3.8% at a concentration of 512 μg/mL. Exposure of porcine intestinal epithelial cells to 16–128 μg/mL rMP1106 caused a 9.5% decrease in cell viability. The rMP1106 was stable over broad ranges of temperature, pH and NaCl concentration. In addition, rMP1106 was highly resistant to papain, proteinase K and snailase digestion but susceptible to trypsin and slightly susceptible to pepsin. These results suggested that rMP1106 has potential as a promising therapeutic drug against S. suis infections.

Keywords
Antimicrobial peptide; MP1106; GAP promoter; Streptococcus suis; Pichia pastoris
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GAP-initiated constitutive expression of a novel plectasin-derived peptide MP1106 by Pichia pastoris and its activity against Streptococcus suis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 50, Issue 2, February 2015, Pages 253–261
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us