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Removal of the N ‐terminal methionine improves the sweetness of the recombinant expressed sweet‐tasting protein brazzein and its mutants in Escherichia coli
Author(s) -
Liu Bo,
Jiang Hua,
Wang Haiyong,
Yang Liu
Publication year - 2021
Publication title -
journal of food biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 47
eISSN - 1745-4514
pISSN - 0145-8884
DOI - 10.1111/jfbc.13354
Subject(s) - sweetness , recombinant dna , chemistry , food science , biochemistry , wine tasting , mutant , escherichia coli , methionine , flavor , amino acid , gene , wine
Abstract The sweet‐tasting protein brazzein is the smallest sweet protein with high sweet potency. Overexpression of this protein in a heterogenous host is an essential way for its production in food industry. In this study, the gene of minor form brazzein was cloned into the pET‐SUMO vector with optimized codon usage and expressed in E. coli BL21‐CodonPlus (DE3)‐RIL. The recombinant protein in absence of the N ‐terminal methionine displayed a sweetness threshold about 1.5 μg/ml, which is the sweetest brazzein protein reported up to now. The unexpected sweet potency of the protein was validated by a series of mutants (7Val → Arg, 9Glu → Lys and 9Glu → Asp), in which E9K exhibited about 50% enhancement of sweetness than the wild type. The superior sweetness of recombinant brazzein and its sweeter mutants suggest their potential applications in food and beverages. Practical applications The sweet‐tasting proteins are natural, low‐, or non‐caloric and nutritive, showing to be promising replacers of sugars and artificial sweeteners, and can be used as sweet additives in the fields of food, medicine, and biotechnology. In the present study, we report that the recombinant brazzein protein expressed in E. coli exhibits superior and improved sweetness than those previously reported. Furthermore, sweeter mutants were obtained with the expression procedure. The superior sweetness of recombinant brazzein and its sweeter mutants suggest their potential applications in food, beverages, and other fields.