Effect of alkyl distribution in pyrazine on pyrazine flavor release in bovine serum albumin solution

Abstract

The flavor release mechanism related to the interaction of aroma compounds with proteins is still unclear. In this study, the interaction of protein with pyrazine homologues, such as 2-methylpyrazine (MP), 2,5-dimethylpyrazine (DP), 2,3,5-trimethylpyrazine (TRP) and 2,3,5,6-tetramethylpyrazine (TEP), was investigated to elucidate the effect of alkyl distribution in a pyrazine ring on its flavor release in bovine serum albumin (BSA) solution (pH 7.4). The results of SPME-GC-MS indicated that methyl distribution in a pyrazine ring significantly affected its release from BSA solution. The pyrazines released from BSA solution with an increasing order of MP, DP, TRP and TEP. The inhibition mechanism of alkyl-pyrazine release was further elucidated by the interaction between alkyl-pyrazines and BSA using multiple spectroscopic methods. The non-covalent interaction between alkyl-pyrazines and BSA was confirmed as the main interaction force by the value of the bimolecular quenching constant (K_q > 2 × 10¹⁰ L mol⁻¹ s⁻¹). A decrease in the hydrophobicity of the microenvironment between the alkyl-pyrazine and BSA was detected by synchronous fluorescence spectra, which revealed that alkyl-pyrazines were mainly bound on the sites of tyrosine and tryptophan in BSA. The UV-vis absorption spectra and circular dichromatic (CD) spectrum revealed that alkyl-pyrazines could induce polarity and conformation change of BSA. The above results indicated that the structure of the flavor homologues can affect their release in food matrices.