Brazzein is an intensely sweet-tasting plant protein with good stability, which makes it an attractive alternative to sucrose. A brazzein gene has been designed, synthesized, and expressed in Escherichia coli at 30 °C to yield brazzein in a soluble form and in considerable quantity. Antibodies have been produced using brazzein fused to His-tag. Brazzein without the tag was sweet and resembled closely the taste of its native counterpart. The brazzein gene was also expressed in Lactococcus lactis, using a nisin-controlled expression system, to produce sweet-tasting lactic acid bacteria. The low level of expression was detected with anti-brazzein antibodies. Secretion of brazzein into the medium has not led to significant yield increase. Surprisingly, optimizing the codon usage for Lactococcus lactis led to a decrease in the yield of brazzein.
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This work was supported by the Slovenian Research Agency Grant No. P4-0127. We are grateful to Prof. Gary M. Dunny for kindly providing the pMSP3545 plasmid and to Prof. Roger Pain for critical reading of the manuscript.
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Berlec, A., Jevnikar, Z., Majhenič, A.Č. et al. Expression of the sweet-tasting plant protein brazzein in Escherichia coli and Lactococcus lactis: a path toward sweet lactic acid bacteria. Appl Microbiol Biotechnol 73, 158–165 (2006). https://doi.org/10.1007/s00253-006-0438-y
- Lactic Acid Bacterium
- Codon Usage
- Sweet Taste
- Alternative Sweetener
- Talon Metal Affinity Resin