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

Abstract:

AIMS:To improve the production of sweet-tasting protein brazzein in Lactococcus lactis using controlled fermentation conditions. METHODS AND RESULTS: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. CONCLUSIONS:Considerable increase in brazzein per cell production was obtained at optimized fermentation conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: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.

journal_name

Lett Appl Microbiol

authors

Berlec A,Tompa G,Slapar N,Fonović UP,Rogelj I,Strukelj B

doi

10.1111/j.1472-765X.2007.02297.x

subject

Has Abstract

pub_date

2008-02-01 00:00:00

pages

227-31

issue

2

eissn

0266-8254

issn

1472-765X

pii

LAM2297

journal_volume

46

pub_type

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