Supplemental material for: Limosilactobacillus fermentum 270 as an adjunct in yak-milk Gouda cheese: Effects on microstructure, microbial succession, and flavor metabolome

Description

Accelerating cheese maturation without compromising quality is a major challenge in plateau dairy processing. This study demonstrates that the Limosilactobacillus fermentum 270 adjunct (LFA) shortens the ripening of yak-milk Gouda cheese by at least 15 days while significantly enhancing its flavor complexity. Using an integrated multi-omics approach (16S rRNA sequencing, untargeted metabolomics, and flavoromics) coupled with the DIABLO framework over a 60-day period, we systematically tracked bacterial, metabolic, and structural dynamics. By day 30, LFA cheeses exhibited advanced proteolysis, accelerated microstructural remodeling, and the premature accumulation of key flavor-active esters and aldehydes. Crucially, we identified a highly efficient, prophage-mediated autolytic mechanism in L. fermentum 270, which triggered the early release of its robust intracellular enzymatic repertoire into the cheese matrix. As confirmed by metabolomics, the activation of this lysis system efficiently drove targeted metabolic shifts, particularly the significant enrichment of arginine biosynthesis and complete galactose utilization. Furthermore, multi-omics integration revealed a distinct cross-omics correlation network, demonstrating how LFA synergizes with the background bacterial community to orchestrate these specific biochemical transformations. Ultimately, this study provides definitive evidence linking strain-specific autolytic and enzymatic traits to the accelerated maturation trajectory, offering a robust strategy for high-quality yak-milk cheese production.

Institutions

• Southwest Minzu University

  Sichuan, Chengdu

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