Tabakaev A V, Tabakaeva O V, Shchelkanov M Yu
Far Eastern Federal University, Institute of Biotechnology, Engineering and Food Systems, 690920, Vladivostok, Russian Island, Ajax, Russian Federation.
G.P. Somov Institute of Epidemiology and Microbiology, Russian Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, 690087, Vladivostok, Russian Federation.
Vopr Pitan. 2023;92(2):43-52. doi: 10.33029/0042-8833-2023-92-2-43-52. Epub 2023 Mar 1.
The creation of specialized food systems capable of correcting body weight and influencing lipid metabolism is an urgent task at the present stage. The main way to create such food systems is to modify product profiles (lipid, protein, carbohydrate, mineral, vitamin, etc.). of the study was to obtain and characterize a new food source of an organic form of chromium for the prevention of hyperlipidemia and obesity in the form of a complex with peptide fractions of fermentolysate proteins of the Mactra chinensis bivalve mollusk from the Far Eastern region. . Soft food parts of the Mactra chinensis bivalve mollusk from the Far Eastern region (the motor muscle and mantle) were used as objects of research. Mollusk specimens were collected in the Amur Bay in June - September 2022. Biotechnological modification of soft tissue proteins was carried out using alkaline protease protozyme B with the following parameters: the duration of the process - 12 and 24 hours, pH - 7.0-7.2, temperature 55 °C. The fractional composition of proteins and peptides was determined by medium pressure gel permeation chromatography, the molecular weight (MW) of proteins and peptides was calculated using MW markers by comparing retention volumes. Extraction of free amino acids was carried out with 70% ethanol for 24 hours at a temperature of 20 °C, the composition and quantitative content of amino acids were determined by HPLC. The complexation reaction was carried out by adding to the fermentolysate with stirring a 10% aqueous solution of CrCl3×6H2O in a mass ratio of 6.25 × nitrogen in the liquid fraction : chromium (III) chloride = 20:1. Process conditions: temperature 20-25 °C, time 60 min, pH 7.0-7.1. The chromium content was determined by the atomic absorption method using spectrophotometer. . The content of low-molecular fractions increased more significantly over time in the mantle fermentolysate. The content of the high-molecular fraction of peptides weighing more than 160 kDa was minimal for both the muscle and the mantle and didn't exceed 1.1%. Low-molecular fractions weighing no more than 18 kDa in the 24-hour fermentolysate accounted for 79.6% for the muscle and 86.9% for the mantle. The obtained fermentolysates were characterized by a high content of the biologically active amino acid taurine - 25.9% (muscle) and 30.05% (mantle) of the total amino acid content. The high content of such essential amino acids as glycine, alanine, leucine, lysine, and the conditionally essential amino acid arginine was also determined. The obtained fermentolysates of soft tissue proteins of the Mactra chinensis were characterized by a fairly balanced amino acid composition. It has been experimentally established that more than 84% of the total chromium content in muscle fermentolysate and 80.9% in mantle fermentolysate was associated with peptide fractions with molecular weights from 24 to 1.4 kDa, and the highest specific chromium content - 1.67 mg/g protein (muscle) and 1.58 mg/g protein (mantle) was determined in the interval fractions with molecular weights of 18.0-12.5 kDa. The minimum specific content of the trace element was established for high-molecular fractions of 160-67 kDa and low-molecular fraction weighing less than 1.4 kDa. . Biotechnological modification of soft tissue proteins of the Mactra chinensis bivalve mollusk from the Far Eastern region made it possible to obtain a balanced amino acid composition fermentolysate with a high content of the free biologically active amino acid taurine. Fortification of the obtained fermentolysate with chromium demonstrated high efficiency of its binding to the amino acid and peptide matrices of the protein hydrolysate. The chromium complex with fermentolysate of proteins of the Mactra chinensis bivalve mollusk can be used as a food source of chromium and taurine, including as a functional ingredient in special food systems for the prevention of hyperlipidemia and obesity.
现阶段,创建能够调节体重并影响脂质代谢的专业食品体系是一项紧迫任务。创建此类食品体系的主要方法是改变产品成分(脂质、蛋白质、碳水化合物、矿物质、维生素等)。本研究的目的是获取并表征一种新型有机形式铬的食物来源,该来源以远东地区中国蛤蜊双壳贝类发酵水解蛋白的肽段复合物形式存在,用于预防高脂血症和肥胖症。远东地区中国蛤蜊双壳贝类的柔软可食部分(运动肌肉和外套膜)被用作研究对象。2022年6月至9月在阿穆尔湾采集了蛤蜊样本。使用碱性蛋白酶原酶B对软组织蛋白进行生物技术修饰,参数如下:过程持续时间为12和24小时,pH值为7.0 - 7.2,温度为55℃。通过中压凝胶渗透色谱法测定蛋白质和肽的分级组成,使用分子量标记物通过比较保留体积计算蛋白质和肽的分子量(MW)。用70%乙醇在20℃下提取游离氨基酸24小时,通过高效液相色谱法测定氨基酸的组成和定量含量。通过在搅拌下向发酵水解产物中加入10%的CrCl₃·6H₂O水溶液进行络合反应,液体部分中氮与氯化铬(III)的质量比为6.25 : 1 = 20 : 1。工艺条件:温度20 - 25℃,时间60分钟,pH值7.0 - 7.1。使用分光光度计通过原子吸收法测定铬含量。随着时间推移,外套膜发酵水解产物中低分子级分的含量增加更为显著。肌肉和外套膜中分子量超过160 kDa的肽的高分子级分含量最低,不超过1.1%。24小时发酵水解产物中分子量不超过18 kDa的低分子级分,肌肉中占79.6%,外套膜中占86.9%。所获得的发酵水解产物的特征在于生物活性氨基酸牛磺酸含量高,分别占总氨基酸含量的25.9%(肌肉)和30.05%(外套膜)。还测定了甘氨酸、丙氨酸、亮氨酸、赖氨酸等必需氨基酸以及条件必需氨基酸精氨酸的高含量。所获得的中国蛤蜊软组织蛋白发酵水解产物的氨基酸组成相当平衡。实验确定,肌肉发酵水解产物中总铬含量的84%以上以及外套膜发酵水解产物中80.9%的铬与分子量为24至1.4 kDa的肽段结合,在分子量为18.0 - 12.5 kDa的区间级分中测定到最高的特定铬含量,分别为1.67 mg/g蛋白质(肌肉)和1.58 mg/g蛋白质(外套膜)。在160 - 67 kDa的高分子级分和分子量小于1.4 kDa的低分子级分中确定了微量元素的最低特定含量。远东地区中国蛤蜊双壳贝类软组织蛋白的生物技术修饰使得能够获得一种氨基酸组成平衡、游离生物活性氨基酸牛磺酸含量高的发酵水解产物。用铬强化所获得的发酵水解产物证明了其与蛋白质水解产物的氨基酸和肽基质结合的高效率。中国蛤蜊双壳贝类蛋白质发酵水解产物的铬复合物可作为铬和牛磺酸的食物来源,包括作为预防高脂血症和肥胖症的特殊食品体系中的功能成分。