Elucidating the role of low molecular weight peptides (< 1 kDa) as Maillard reactant flavour precursors in selected cooked white and red meats using a flavouromics approach

使用风味组学方法阐明低分子量肽 (< 1 kDa) 作为美拉德反应物风味前体在选定的熟白肉和红肉中的作用

• 批准号: RGPIN-2014-04815
• 负责人: Aliani, Michel
• 金额: $ 1.89万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2017
• 资助国家: 加拿大
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=630199
• 关键词: Elucidating; role; low; molecular; weight

Meat flavour is formed upon heating via chemical reactions between water and fat-soluble flavour precursors present in raw meat. Although free amino acids, reducing and phosphorylated sugars play a major role via the Maillard reaction, the role of small peptides as Maillard reactants in meat and related products need to be elucidated. The role of low MW peptides as key flavour precursors in various types of meat, their formation, identification and contribution to the generation of key VOCs with a significant impact on cooked meat aroma and flavour will be investigated. Meat extracts (chicken breast, thigh, beef and bison) will be prepared using a developed water extraction method, filtered using a 1 K Da cutoff membrane size and the resulting filtrate will be analyzed using a recently developed high resolution liquid chromatography mass spectrometry technique to identify and quantify all small size compounds present in raw meat. Model systems containing natural concentrations of identified peptides, reducing sugars and amino acids (purchased or synthesized) will be used to investigate the Maillard products generated upon heating. Liquid chromatography (LC) quadruple time of flight (QTOF) Mass Spectrometry (MS) and gas chromatography-MS/MS will be used to identify the intermediary compounds (non and semi-volatiles) and the VOCs generated during heating. Stable isotope amino acids, peptides and sugars (purchased or synthesized) will be used to elucidate the contribution of individual Maillard precursors to generation of key VOCs. Sensory evaluation of model systems and meat samples with added selected peptides will be investigated to further clarify the specific role played by each peptide. Multiple regression statistical analysis and partial least squares (PLS) statistical tests will be performed to elucidate the relationship between peptides, reducing sugars, amino acids and the sensory attributes generated by trained sensory panels. Specific objectives are to: (1) Investigate the formation, identification and quantification of low molecular weight (MW) peptides (< 1 KDa) in meat extracts (chicken breast and thigh, beef, bison and fish) using high resolution mass spectrometry techniques (2) Identify and quantify the volatile organic compounds (VOCs) generated in a model system containing natural concentrations of selected peptides and key reducing and phosphorylated sugars using stable isotope dilution mass spectrometry techniques. (3) Elucidate the individual contribution of selective small peptides to the overall acceptability of cooked meat aroma and flavour in selected meat species using consumer acceptability tests. (4) Determine the sensory attributes generated with added selected peptides to raw meat prior to cooking using a trained sensory panel and (5) Clarify the effect of different cooking methods on data obtained in objectives 1 to 4. Novelty of proposal: Natural proteolytic enzymes are active in muscle post-mortem with a continuous generation of small size peptides over time resulting in a significant increase in meat tenderness and availability of small size peptides as Maillard reactants with a potential significant impact on aroma and flavour. Therefore, this proposal is highly novel in seeking to provide data to elucidate the role played by low MW peptides as Maillard reactants in flavour formation in meat. Significance: Proteolytic enzymes are abundant in many foods generating hundreds of low MW peptides that can act as NH2 donor in the Maillard reaction. A proper understanding of the mechanism by which peptides affect flavour in meat and other complex foods could enable the development of acceptable nutritional formulations containing peptides with selective functional properties.

肉味是在加热时通过水和生肉中存在的脂溶性风味前体之间的化学反应形成的。尽管游离氨基酸、还原糖和磷酸化糖在美拉德反应中发挥着重要作用，但小肽作为美拉德反应物在肉类和相关产品中的作用仍需阐明。将研究低分子量肽作为各种肉类中关键风味前体的作用、它们的形成、识别以及对关键挥发性有机化合物的产生的贡献，这些挥发性有机化合物对熟肉的香气和风味有重大影响。肉类提取物（鸡胸肉、大腿肉、牛肉和野牛）将使用先进的水提取方法制备，使用 1 K Da 截止膜尺寸进行过滤，所得滤液将使用最近开发的高分辨率液相色谱质谱技术进行分析，以识别和量化生肉中存在的所有小尺寸化合物。含有天然浓度的已识别肽、还原糖和氨基酸（购买或合成）的模型系统将用于研究加热时产生的美拉德产品。液相色谱 (LC) 四极飞行时间 (QTOF) 质谱 (MS) 和气相色谱-MS/MS 将用于鉴定中间化合物（非挥发性和半挥发性）以及加热过程中产生的 VOC。稳定同位素氨基酸、肽和糖（购买或合成）将用于阐明各个美拉德前体对关键 VOC 生成的贡献。将研究添加选定肽的模型系统和肉类样品的感官评价，以进一步阐明每种肽所发挥的具体作用。将进行多元回归统计分析和偏最小二乘（PLS）统计测试，以阐明肽、还原糖、氨基酸和经过训练的感官小组产生的感官属性之间的关系。具体目标是： (1) 使用高分辨率质谱技术研究肉类提取物（鸡胸肉和大腿、牛肉、野牛和鱼）中低分子量 (MW) 肽 (< 1 KDa) 的形成、鉴定和定量 (2) 鉴定和定量模型系统中产生的挥发性有机化合物 (VOC)，该模型系统包含天然浓度的选定肽以及关键还原和磷酸化肽 使用稳定同位素稀释质谱技术测定糖。 (3) 通过消费者可接受性测试，阐明选择性小肽对选定肉类品种的熟肉香气和风味的总体可接受性的个体贡献。 (4) 使用训练有素的感官小组确定在烹饪前向生肉中添加选定的肽所产生的感官属性，以及 (5) 阐明不同烹饪方法对目标 1 至 4 中获得的数据的影响。提案的新颖性：天然蛋白水解酶在死后肌肉中具有活性，随着时间的推移不断产生小尺寸肽，导致肉的嫩度和小尺寸肽作为美拉德反应物的可用性显着增加 对香气和风味有潜在的重大影响。因此，该提案在寻求提供数据来阐明低分子量肽作为美拉德反应物在肉中风味形成中所发挥的作用方面具有高度新颖性。意义：许多食物中富含蛋白水解酶，可产生数百种低分子量肽，这些肽可在美拉德反应中充当 NH2 供体。正确了解肽影响肉类和其他复杂食品风味的机制，可以开发出包含具有选择性功能特性的肽的可接受的营养配方。