Distinct amino acid synthesis constraints regulate aroma ester formation under carbon–nitrogen adjustment in Cyberlindnera fabianii

Abstract

Non-Saccharomyces yeasts significantly contribute to aroma complexity in fermented foods and beverages; however, the role of nutrient availability in constraining ester biosynthesis remains poorly understood. Here, an aroma-active non-Saccharomyces yeast, Cyberlindnera fabianii, isolated from a traditional fermented food, was used to investigate how carbon availability and carbon-to-nitrogen (C/N) ratio regulate ester biosynthesis. Five nutritional conditions were examined by integrating gas chromatography–mass spectrometry (GC–MS)–based volatile profiling, transcriptomics, and targeted intracellular metabolomics. High carbon and medium-to-low C/N ratios enhanced ester production, with phenethyl acetate and isoamyl acetate as major discriminant compounds. In contrast, low carbon and high C/N ratios favored organic acid accumulation and off-flavor formation. Multivariate analyses revealed ester-specific metabolic constraints: phenethyl acetate synthesis was limited by aromatic amino acid supply, whereas isoamyl acetate depended on branched-chain amino acid biosynthesis. Overall, these results demonstrate that carbon availability and C/N ratio reposition metabolic bottlenecks in ester formation, providing a nutrition-based framework for aroma modulation.

Keywords

Non-Saccharomyces yeast / Amino acid metabolism / C/N ratio / Metabolic bottlenecks / Ester biosynthesis

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Funding

Key Technologies Research and Development Program(2021YFC2102000)

National Natural Science Foundation of China(22178147)

Postdoctoral Fellowship Program of CPSF(GZB20250577)

Program of Introducing Talents of Discipline to Universities(111-2-06)

High-end Foreign Experts Recruitment Plan of China(G2021144005L)

National Program for Support of Top-notch Young Professionals

Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions

Top-notch Academic Programs Project of Jiangsu Higher Education Institutions

Jiangsu province "Collaborative Innovation Center for Advanced Industrial Fermentation" industry development program

Basic Research Program of Jiangsu Province

Jiangsu Basic Research Center for Synthetic Biology(No. BK20233003)

RIGHTS & PERMISSIONS

Jiangnan University