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. 2024 Sep 18:15:1464953.
doi: 10.3389/fmicb.2024.1464953. eCollection 2024.

Unlocking the potential of Kluyveromyces marxianus in the definition of aroma composition of cheeses

Giorgia Perpetuini  1 , Alessio Pio Rossetti  1 , Arianna Rapagnetta  1 , Rosanna Tofalo  1
Affiliations

Unlocking the potential of Kluyveromyces marxianus in the definition of aroma composition of cheeses

Giorgia Perpetuini et al. Front Microbiol. .

Abstract

Introduction: The cheese microbiota is very complex and is made up of technologically-relevant, spoilage, opportunistic and pathogenic microorganisms. Among them lactic acid bacteria and yeasts are the main ones. One of the most interesting dairy yeasts is Kluyveromyces marxianus because of its technological properties including the ability to produce aroma compounds.

Methods: This study investigated the contribution of Kluyveromyces marxianus to the gross composition and aroma profile of cow cheeses. Experimental cheeses were prepared by inoculating a co-culture of K. marxianus FM09 and a commercial strain of Lacticaseibacillus casei and compared with cheeses obtained with only L. casei. The gross composition was determined by a FoodScan™ 2 Dairy Analyser, and free amino acids were evaluated at 507 nm after reaction with Cd-ninhydrin. The volatile organic compounds were extracted by head-space solid phase micro-extraction and analyzed by gas chromatography-mass spectrometry coupled with odor activity values. qRT-PCR was applied to determine the expression of genes involved in esters synthesis and degradation.

Results: The inoculation of K. marxianus induced an increase of pH and a reduction of protein content of cheeses, in agreement with the stronger proteolysis detected in these cheeses. K. marxianus influenced the content of aroma compounds both quantitatively and qualitatively. In particular, an increase of higher alcohols, esters and organic acids was observed. Moreover, 12 compounds were detected only in cheeses obtained with the co-culture. These differences were in agreement with the odor activity values (OAV). In fact, only 11 compounds showed OAV > 1 in cheeses obtained with the commercial strain, and 24 in those obtained with the co-culture. The qPCR analysis revealed an over expression of ATF1, EAT1, and IAH1 genes.

Conclusion: Kluyveromyces marxianus could act as an important auxiliary starter for cheese production through the development and diversification of compounds related to flavor in short-aged cow cheeses.

Keywords: Kluyveromyces marxianus; OAV; aroma compounds; cheese; qPCR.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.

Figures

Figure 1
Figure 1
Evolution of free amino acids throughout ripening in cheeses produced with the commercial starter (CC) and CC + K. marxianus FM09. Data are means ± standard deviations of 3 independent experiments, each carried out in triplicate. * p<0.05.
Figure 2
Figure 2
Evolution of lactic acid bacteria and yeasts in the cheeses obtained with CC (A) and CC + FM09 (B). Data are means ± standard deviations of 3 independent experiments, each carried out in triplicate.
Figure 3
Figure 3
Venn diagram showing the similarities and differences of volatile compounds between cheese samples The Venn diagram was generated with the web tool provided by the Bioinformatics and Systems Biology of Gent.
Figure 4
Figure 4
Relative transcript levels of genes involved in ester content regulation in K. marxianus after 30 days of ripening of cheeses made with CC + FM09. Transcript levels are expressed as x-fold increase in comparison with the expression at T0. Three biological replicates were performed, with 3 technical repeats.
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The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. The publishing fees were supported by University of Teramo.

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