The AGPL1 (ADP-glucose pyrophosphorylase large subunit 1) promoter from watermelon (Citrullus vulgaris S.) has proved to exhibit fruit-specific expression patterns in tomato (Lycopersicon esculentum L.). A plant expression vector harboring sweet-taste protein, Brazzein, directed by AGPL1 promoter, was constructed and transferred into tomato plants through Agrobacterium-mediated transform methods. Histochemical staining assay, PCR screening, Southern blotting analysis and RT-PCR analysis showed that Brazzein gene was successfully integrated into the genome of transgenic tomato plants with stable expression. Sweet-taste fruits were produced under control of fruit-specific AGPL1 promoter, whereas other parameters of fruit quality were largely unchanged.

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http://dx.doi.org/10.3724/sp.j.1005.2009.00663DOI Listing

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Article Synopsis
  • Sweeteners are commonly used to reduce sugar intake in diets, but they often don't taste as good as natural sugars, leading to the creation of a new sweet protein called brazzein, produced from the strain Komagataella phaffii CF-st401.* -
  • The research focused on assessing the safety of this genetically modified microorganism (K. phaffii CF-st401) using in silico methods, analyzing its plasmid and DNA structure for potential risks like toxin production, antibiotic resistance, and allergenicity.* -
  • Findings showed that over 70% of the genetic material responsible for producing brazzein in K. phaffii CF-st401 is similar to the reference protein from the plant Pent
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Novel gnd_v2 Fusion Tag and Engineered TEV Protease Enable Efficient Production of Brazzein.

J Microbiol Biotechnol

November 2024

CCZU-JITRI joint Bio-X Lab, School of Pharmacy & School of Biological and Food Engineering, Changzhou University, 213164, Changzhou, Jiangsu Province, P.R. China.

Protein solubility and purification challenges often hinder the large-scale production of valuable proteins like brazzein, a potent sweet protein with significant health benefits and commercial potential. This study introduces two novel tools to overcome protein expression and purification bottlenecks: a gnd_v2 fusion tag and an engineered Tobacco Etch Virus (TEV) protease. The gnd_v2 tag, derived from 6-phosphogluconate dehydrogenase, was engineered to improve the soluble expression of brazzein.

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The human palate can discern multiple tastes; however, it predominantly perceives five fundamental flavors: sweetness, saltiness, sourness, bitterness, and umami. Sweetness is primarily mediated through the sweet taste receptor, a membrane-bound heterodimeric structure comprising T1R2-T1R3. However, unraveling the structural and mechanistic intricacies of the sweet taste receptor has proven challenging.

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Sweet-tasting proteins (SPs) are proteins of plant origin initially isolated from tropical fruits. They are thousands of times sweeter than sucrose and most artificial sweeteners. SPs are a class of proteins capable of causing a sweet taste sensation in humans when interacting with the T1R2/T1R3 receptor.

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The excessive consumption of sugar-containing foods contributes to the development of a number of diseases, including obesity, diabetes mellitus, etc. As a substitute for sugar, people with diabetes mellitus and obesity most often use sweeteners. Sweet proteins, in particular brazzein, are an alternative to synthetic sweeteners that have natural origin, are broken down in the intestines along with food proteins, and do not affect blood sugar and insulin levels.

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