Beer quality is largely guaranteed by a rational ratio of different flavour compounds. In this study, we present intracellular metabolic profiles in the light of lager beer flavour compound synthesis on an industrial scale. A total of 62 intracellular metabolites were identified and quantitatively analysed. Among these, amino acid was identified as the most dominant category of metabolites. We also report gene expression profiling of relevant genes encoding six representing amino acids, including serine, alanine, lysine, valine, leucine and asparagine. Nine genes involved in the amino acid synthesis – ALT1, ALT2, ASN1, ASN2, BAT1, SER2, SHM1, SHM2 and LYA1 – displayed significantly higher level of expression at various fermentation stages in an industrial lager fermentation. However, concentrations of the recovered amino acids decreased towards the end of fermentation. Our results suggested that the defined amino acids could be assimilated by the yeast to be utilized for synthesis of aroma‐active metabolites at different stages in fermentation. Copyright © 2017 The Institute of Brewing & Distilling