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ABSTRACT: The objective of this study was to examine the aroma profiles of 12 Gouda cheeses sold in China and to determine which aromas were preferred by young Chinese consumers (n = 110). The consumers selected 11 descriptors of the aromas of the Gouda cheeses in a check-all-that-apply questionnaire. These 11 descriptors were used by a panel of experts for sensory analysis to perform a quantitative descriptive analysis of the cheeses. A principal component analysis of the data from the quantitative descriptive analysis revealed that the characteristic aromas of young Gouda cheeses, medium-aged Gouda cheeses, and aged Gouda cheeses were “milky” and “whey”; “creamy” and “sour”; and “rancid” and “nutty,” respectively. The results of a penalty analysis combined with the check-all-that-apply results and the preference scores showed that the 3 groups of young Chinese consumers (those who often ate cheese, occasionally ate cheese, and never ate cheese) preferred the Gouda cheeses with “milky” or “creamy” aromas and did not enjoy those with “sour” or “rancid” aromas. Occasional cheese eaters comprised the majority of the young Chinese consumers, and they were more tolerant of the Gouda cheeses with “whey” and “sulfury” aromas than those who often or never ate cheese. In addition, we identified a positive correlation between the consumers' preferences for the aromas of the Gouda cheeses and their willingness to pay for them. Overall, the results of this study should help promote the development of Gouda cheeses and associated products that meet the preferences of young Chinese consumers.

ABSTRACT: Milk fan is an acid-curd cheese with strong national characteristics (a traditional dairy product of the Bai nationality with a shape like a piece of paper) and a long history in Yunnan province, China. In our previous study, we characterized the microbial community diversity of milk fan, but the succession of microorganisms associated with flavor formation in milk fan is still unknown. Therefore, we examined the predominant microorganisms and their correlations with the formation of flavor in the fermentation of sour juice and drying of milk fan by gas chromatography mass spectrometry, high-throughput 16S rDNA sequencing, intergenic spacer sequencing and metatranscriptome analysis. We found that the relative abundances of Lactobacillus and Issatchenkia initially decreased and then increased with time during the fermentation of sour juice. However, the relative abundances of Acetobacter, Leuconostoc, Lactococcus, Geotrichum, and Dipodascus initially increased and then decreased. During the drying step, the relative abundances of Lactobacillus and Issatchenkia continuously increased and became the dominant microorganisms in the milk fan. The metatranscriptomes generated from the milk fan showed that “carbohydrate metabolism,” “translation,” and “signal transduction” were the main metabolic functions of the microbial communities. Rhodotorula and Yarrowia contained more differentially expressed genes than other genera, which indicated they may be associated with the production of the characteristic flavor. Furthermore, a Pearson correlation analysis showed that Lactococcus, Rhodotorula, Candida, Cutaneotrichosporon, and Yarrowia were significantly positively correlated with more aroma-active compounds, mainly ethyl acetate, 2-heptanone, isovaleraldehyde, butyric acid, nonanal, and hexanal. In conclusion, these findings contribute to a better understanding of the flavor production mechanism during the production of milk fan.

ABSTRACT: Diacetyl and acetoin are key aroma components of fermented milk but are produced in low concentrations by starter cultures. In this study, we expressed NADH oxidase, acetolactate synthase, and inactivated acetolactate decarboxylase in Lacticaseibacillus casei TCS to generate recombinant L. casei strains, and investigated the effects of the genes encoding these enzymes on diacetyl and acetoin production during milk fermentation. In the single-gene recombinant strains tested, diacetyl concentrations were highest in milk fermented by L. casei TCSI-nox (nox gene overexpressed, 3.68 mg/kg), whereas acetoin concentrations were highest in milk fermented by L. casei TCS-ΔalsD (alsD gene deleted, 32.94 mg/kg). Moreover, diacetyl and acetoin concentrations were higher in the inducible strains than in the corresponding constitutive strains (e.g., TCSI-nox vs. TCSC-nox, and TCSI-ΔalsD-nox vs. TCSC-ΔalsD-nox). This phenomenon was also reflected in the protein expression levels and enzyme activities. In the double-gene recombinant strains tested, the highest concentrations of diacetyl and acetoin were produced by L. casei TCSI-ΔalsD-nox (nox overexpressed and alsD deleted, 4.66 mg/kg, 69.62 mg/kg, respectively). The triple-gene recombinant L. casei TCS-ΔalsD-nox-alsS produced the highest concentrations of diacetyl and acetoin, which were 2.38 and 11.19 times, respectively, the concentrations produced by the original strain. These results show that the nox, alsS, and alsD genes make key contributions to the biosynthesis of diacetyl and acetoin by L. casei. The modification of multiple genes had a synergistic effect, leading to greatly increased synthesis of diacetyl and acetoin by L. casei during its fermentation of milk.

ABSTRACT: 3-Methylbutanal is a key volatile compound that imparts a nutty flavor to Cheddar cheese. Lactococcus lactis has been successfully applied as a starter to increase the level of 3-methylbutanal produced during the ripening of cheese. However, the mechanism of action and genetic diversity of this bacterium for 3-methylbutanal biosynthesis remains unclear. In this study, we investigated the association between the L. lactis genotype and phenotype in the biosynthesis of 3-methylbutanal via both direct and indirect pathways. Fourteen strains of L. lactis were screened for the capacity to produce 3-methylbutanal, and strain 408 (>140 μM) produced the highest among all tested strains, which exhibited both α-keto acid decarboxylase and α-ketoacid dehydrogenase activities. Furthermore, the results of a sodium meta-arsenite inhibition experiment showed that the 3-methylbutanal–producing capacities of each strain declined to various degrees. The kdcA gene, which encodes the direct pathway component α-ketoacid decarboxylase, was detected in 4 of the 14 strains, of which only strain 408 contained the full-length gene. We then characterized the genes associated with the indirect pathway by detecting the expression levels of the pdh gene cluster, ack, and pta, which were expressed at relatively higher levels in a high-yield strain than in a low-yield strain. As a result, these L. lactis strains were divided into 3 categories according to gene diversity, gene expression, and 3-methylbutanal production. The results of this study refine our knowledge of the genetic determinants of 3-methylbutanal biosynthesis in L. lactis and explain the effect of both synthesis pathways on 3-methylbutanal production.