Markova E V, Leonova E I, Sopova Ju V
St. Petersburg State University, 199034, St. Petersburg, Russian Federation.
St. Petersburg Branch, Vavilov Institute of General Genetics, 199034, Saint Petersburg, Russian Federation.
Vopr Pitan. 2024;93(1):61-71. doi: 10.33029/0042-8833-2024-93-1-61-71. Epub 2024 Jan 19.
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. of the review was to analyze the available data on the sweet protein brazzein, its physical and chemical properties, existing biotechnological methods of production, and prospects for application in the food industry in order to further develop an optimized heterologous expression system. . Google Scholar, Scopus, Web of Science, PubMed, RSCI and eLibrary.ru databases were used for collecting and analyzing literature. Search depth - 30 years. . Numerous studies of the physical and chemical properties of brazzein have demonstrated its high potential for use in the food industry. In particular, a short amino acid sequence, thermal stability, the ability to maintain its structure and sweet properties in a wide pH range, hypoallergenicity, lack of genotoxicity, and an extremely high level of sweetness compared to sucrose allow us to conclude that its use is promising. Mutant variants of brazzein have been generated, the sweetest of which (with three amino acid substitutions H31R/E36D/E41A) exceeds sucrose sweetness by 22 500 times. To date, various systems for the expression of recombinant brazzein have already been developed, in which bacteria (Escherichia coli, Lactococcus lactis, Bacillus licheniformis), yeast (Komagataella phaffii, Kluyveromyces lactis, Saccharomyces cerevisiae), plants (Zea mays, Oryza sativa, Lactuca sativa, Nicotiana tabacum, Daucus carota) and animals (Mus musculus) have been used. . Due to its high sweetness, organoleptic properties and long history of human consumption, brazzein can be considered as a promising natural sweetener. Despite the short peptide sequence, the production of the recombinant protein faced a number of problems, including low protein yield (for example, it could only be detected in mouse milk by Western blot hybridization) and loss of sweetness. Thus, further optimization of the process is necessary for widespread brazzein use in the food industry, which includes the selection of an adequate producer and the use of extracellular expression systems to reduce the final cost of the product.
过量食用含糖食物会导致多种疾病的发生,包括肥胖症、糖尿病等。作为糖的替代品,糖尿病患者和肥胖者最常使用甜味剂。甜蛋白,尤其是布拉齐因,是合成甜味剂的一种天然替代品,它会在肠道内与食物蛋白一起被分解,不会影响血糖和胰岛素水平。本综述的目的是分析关于甜蛋白布拉齐因的现有数据、其物理和化学性质、现有的生物技术生产方法以及在食品工业中的应用前景,以便进一步开发优化的异源表达系统。利用谷歌学术、Scopus、科学网、PubMed、俄罗斯科学引文索引(RSCI)和eLibrary.ru数据库收集和分析文献。搜索深度为30年。众多关于布拉齐因物理和化学性质的研究表明其在食品工业中有很高的应用潜力。特别是,其氨基酸序列短、热稳定性好、在较宽的pH范围内能保持其结构和甜味特性、低过敏性、无遗传毒性,并且与蔗糖相比甜度极高,这些特性使我们得出其应用前景广阔的结论。已经产生了布拉齐因的突变变体,其中最甜的变体(有三个氨基酸替换H31R/E36D/E41A)的甜度比蔗糖高出22500倍。迄今为止,已经开发出了各种用于表达重组布拉齐因的系统,其中使用了细菌(大肠杆菌、乳酸乳球菌、地衣芽孢杆菌)、酵母(毕赤酵母、乳酸克鲁维酵母、酿酒酵母)、植物(玉米、水稻、生菜、烟草、胡萝卜)和动物(小家鼠)。由于其高甜度、感官特性以及人类食用历史悠久,布拉齐因可被视为一种有前景的天然甜味剂。尽管其肽序列较短,但重组蛋白的生产面临一些问题,包括蛋白产量低(例如,通过蛋白质免疫印迹杂交只能在小鼠乳汁中检测到)以及甜味丧失。因此,为了使布拉齐因在食品工业中广泛应用,有必要进一步优化生产工艺,这包括选择合适的生产宿主以及使用细胞外表达系统来降低产品的最终成本。