Optimization of fermentation conditions for the expression of sweet-tasting protein brazzein in Lactococcus lactis

Letters in Applied Microbiology
Ales BerlecBorut Strukelj

Abstract

To improve the production of sweet-tasting protein brazzein in Lactococcus lactis using controlled fermentation conditions. The nisin-controlled expression system was used for brazzein expression. The concentration of nisin for induction and the optical density (OD) at induction were therefore optimized, together with growth conditions (medium composition, pH, aerobic growth in the presence of hemin). Brazzein was assayed with ELISA on Ni-NTA plates and Western blot. Use of the M-17 medium, containing 2.5% glucose, anaerobic growth at pH 5.9 and induction with 40 ng ml(-1) nisin at OD 3.0 led to an approx. 17-fold increase in brazzein per cell production compared to non-optimized starting conditions. Aerobic growth in the presence of hemin did not increase the production. Considerable increase in brazzein per cell production was obtained at optimized fermentation conditions. Optimized growth conditions could be used in application of brazzein expression in L. lactis. The importance of pH and OD at induction contributes to the body of knowledge of optimal recombinant protein expression in L. lactis. The new assay for brazzein quantification was introduced.

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Citations

Oct 4, 2012·Preparative Biochemistry & Biotechnology·Few Ne ChewBeng Ti Tey
Jan 1, 2012·SpringerPlus·Aleksandra M MirończukMarcin Lukaszewicz
Mar 6, 2012·Annual Review of Food Science and Technology·Martin B PedersenAlexandra Gruss
Dec 5, 2014·Microbial Cell Factories·Olivia Cano-GarridoElena García-Fruitós
Jan 8, 2011·Current Opinion in Biotechnology·Delphine LechardeurAlexandra Gruss
Aug 5, 2015·Applied Microbiology and Biotechnology·Christopher D Skory, Gregory L Côté
Feb 5, 2015·World Journal of Microbiology & Biotechnology·Wenliang GuXinwen Hu
Jan 28, 2021·Bioprocess and Biosystems Engineering·Se-Woong ParkKwang-Hoon Kong
Sep 11, 2012·Journal of Agricultural and Food Chemistry·Nicolas PoirierLoïc Briand

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