کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5132782 1492055 2017 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
An improved process for the production of highly purified recombinant thaumatin tagged-variants
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
An improved process for the production of highly purified recombinant thaumatin tagged-variants
چکیده انگلیسی


- Co-expression of thaumatin and a molecular chaperone increases secretion 2-fold.
- An optimised purification methodology of recombinant thaumatin is reported.
- Production of highly purified recombinant thaumatin variants is detailed.
- Thaumatin variants for thaumatin:receptor interaction studies have been produced.

The sweetest tasting molecule known is the protein thaumatin, first isolated from the katemfe fruit, Thaumatococcus daniellii. Thaumatin is used in the food and beverage industry as a low-calorie sugar substitute. Thaumatin interacts with taste receptors in the oral cavity eliciting a persistent sweet taste and a bitter, liquorice flavor. Recombinant thaumatin was expressed in Pichia pastoris and through a co-expression strategy with a molecular chaperone, yields of one engineered thaumatin variant increased by greater than two-fold. A detailed purification strategy for thaumatin is reported resulting in a homogenous sample recovered at a yield of 42%. The recombinant thaumatins were extensively characterised using size exclusion chromatography for homogeneity, reversed-phase HPLC for purity (99%), peptide digest LC-MS/MS for sequence determination, and circular dichroism and tryptophan fluorescence spectroscopies for conformational characterisation. These new thaumatin variants are amenable for bioconjugation, providing chemical biology tools for thaumatin:taste receptor interaction studies.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Food Chemistry - Volume 237, 15 December 2017, Pages 825-832
نویسندگان
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