Search Results (53)

Calcareous soils, characterized by high pH and calcium carbonate content, often limit the availability of essential nutrients for crops such as rice (Oryza sativa L.), reducing yield and nutritional quality. In this study, we evaluated the effect of the halotolerant yeast Debaryomyces hansenii on the growth, nutrient uptake, and phosphorus acquisition mechanisms of rice plants cultivated in calcareous soil under controlled greenhouse conditions. Plants inoculated with D. hansenii, particularly via root immersion, exhibited significantly higher SPAD chlorophyll index, plant height, and grain yield compared to controls. A modest increase (~4%) in dry matter content was also observed under sterilized soil conditions. Foliar concentrations of Fe, Zn, and Mn significantly increased in plants inoculated with D. hansenii via root immersion in non-sterilized calcareous soil, indicating improved micronutrient acquisition under these specific conditions. Although leaf phosphorus levels were not significantly increased, D. hansenii stimulated acid phosphatase activity, as visually observed through BCIP staining, and upregulated genes involved in phosphorus acquisition under both P-sufficient and P-deficient conditions. At the molecular level, D. hansenii upregulated the expression of acid phosphatase genes (OsPAP3, OsPAP9) and a phosphate transporter gene (OsPTH1;6), confirming its influence on P-related physiological responses. These findings demonstrate that D. hansenii functions as a plant growth-promoting yeast (PGPY) and may serve as a promising biofertilizer for improving rice productivity and nutrient efficiency in calcareous soils, contributing to sustainable agricultural practices in calcareous soils and other nutrient-limiting environments. Full article

The metabolic activity of fermentative microorganisms plays a critical role in determining the flavor profile of fermented meat products. Modulating carbon and nitrogen sources represents a promising strategy for enhancing product quality. In this study, Debaryomyces hansenii strains isolated from dry-cured ham were assessed in a sterile sausage model to evaluate the effects of different carbon sources (sucrose, corn starch) and nitrogen sources (leucine, soy protein isolate) on colony growth, enzyme activity, and physicochemical properties. These nutritional factors significantly affected the fermentation performance of D. hansenii. Corn starch and soy protein isolate increased colony count by 14.94% and 90%, respectively, and enhanced protease activity by 2-fold and 4.5-fold. Both treatments maintained high lipase activity (>50 U/g). Both supplements improved the water-holding capacity and decreased the water activity. Carbon sources reduced the medium pH, whereas nitrogen sources contributed to the maintenance of pH stability. A further analysis indicated that corn starch promoted the accumulation of aldehydes and ketones, which intensified the sourness and suppressed the saltiness. In contrast, soy protein isolate increased the abundance of free amino acids associated with umami and sweetness, and stimulated the formation of esters, ketones, and pyrazines, thereby enhancing flavor richness and umami intensity. Both ingredients also reduced saturated fatty acid levels and increased the unsaturated to saturated fatty acid ratio. Soy protein isolate exhibited a more pronounced effect on D. hansenii fermentation. This study provides a technical reference for enhancing the flavor characteristics of fermented meat products via the adjustment of carbon and nitrogen sources to regulate D. hansenii fermentation. Full article

Throughout the process of dry-cured ham, moulds such as P. nordicum, a producer of ochratoxin A (OTA), grow on its surface. The use of combined biocontrol agents (BCAs) is a promising strategy for controlling this hazard. The goal of this study is to assess the effect of D. hansenii, S. xylosus, and P. chrysogenum as BCAs on the metabolome of two strains of P. nordicum and to understand the differences between both strains. Each ochratoxigenic strain was inoculated both individually and in combination with the BCAs onto ham for 30 days under the environmental conditions experienced during traditional ripening. Untargeted metabolomics was performed through mass spectrometry using a Q-Exactive Plus Orbitrap. The BCAs caused alterations in the metabolomes of both ochratoxigenic moulds, mainly in phenylalanine catabolism and the valine, leucine, and isoleucine biosynthesis pathways, although with some differences. In the absence of the BCAs, the metabolomes of both types of P. nordicum were globally changed, despite these being moulds of the same species. In conclusion, these data help us to understand the differences between OTA-producing strains in dry-cured ham and confirm the need to demonstrate the efficacy of BCAs against a wide range of toxigenic moulds before they can be used to minimise OTA contamination in the meat industry. Full article

The rejection of chemical preservatives reflects the growing demand for natural and safe products. This concern has spurred scientific interest in yeasts as biocontrol agents, given their antagonistic activity against undesired fungi, which is one of the main problems associated with preservative reduction. Debaryomyces hansenii is a non-conventional yeast that has shown great potential for inhibiting filamentous fungi in the food industry. This study investigated the role of nutrient competition in the biocontrol activity of D. hansenii against unwanted molds. Potentially pathogenic molds from spoiled food were isolated using different media and identified using Sanger sequencing. The inhibitory effects of different autochthonous D. hansenii strains under varying nutrient conditions were assessed against isolated molds using semipermeable membranes. Inhibition activity was measured by assessing mycelial expansion and spore production using image software analysis and classical cell counting using a Neubauer chamber. The results indicated that D. hansenii effectively inhibited mold growth and sporulation, with the autochthonous strains LR2 and SRF1 showing higher inhibitory activity than the control strain CBS767. The effectiveness of inhibition varied with the yeast–mold combination, highlighting the need for a species-specific analysis. Nutrient competition plays a complementary role in D. hansenii biocontrol but does not directly impact overall inhibition. This suggests that other mechanisms, such as direct cell interactions or metabolite production, may be crucial. These findings enhance our understanding of the potential of D. hansenii as a natural preservative and advance biocontrol methods for food safety. Full article

Debaryomyces hansenii naturally colonize wheat grain and can potentially inhibit the pathogens responsible for Fusarium crown rot (FCR). Seed dressing is a recommended method for protecting crops against FCR pathogens. The effectiveness of seed dressing with antagonistic yeasts in reducing the incidence of FCR remains insufficiently investigated. The aim of this study was to evaluate the effect of seed dressing with a triazole fungicide and a suspension of D. hansenii cells on the health status and development of durum wheat cultivars (Durasol and Floradur), and to analyze the structure of the mycobiome in the rhizosphere of seedlings. Under field conditions, the incidence of FCR was reduced by 57.1% by triticonazole and 35.7% by the biocontrol agent relative to the control treatment. Seed dressing with D. hansenii decreased the number of operational taxonomic units (OTUs) of Fusarium pathogens by 47.24% in cv. Durasol and 87.4% in cv. Floradur. The number of OTUs of autochthonous yeast species and Mortierellomycota increased in the rhizosphere of both durum wheat cultivars. The effectiveness of seed dressing with yeasts is determined by the quality and local adaptation of biocontrol agents. Full article

Dry-aged beef has gained interest worldwide in recent years due to its improved sensory attributes. This enhancement is thought to be partially driven by microbial activities, particularly lipolysis and proteolysis. In this study, dry-aged beef manufactured by seven commercial producers in China was analyzed. The pH value and total volatile base nitrogen value of dry-aged beef were determined. High-throughput amplicon sequencing of full-length 16S rRNA genes and internal transcribed spacer (ITS) regions was used to analyze the microbial community. A total of 207 proteolytic and lipolytic isolates were identified by sequencing 16S rRNA genes for bacteria and sequencing the D1/D2 region of 28S rRNA genes and the ITS region for fungi. The results revealed that the crust harbored greater numbers of bacteria and fungi than the interior. The bacterial community was dominated by Pseudomonas species, which were core members in both the crust and interior, while Brochothrix thermosphacta was identified as a core bacterium exclusively in the crust. The fungal community primarily constituted Candida sake, Kurtzmaniella species, and members of the phylum Chytridiomycota. Proteolytic and lipolytic isolates were mainly identified as Pseudomonas sp., B. thermosphacta, Carnobacterium maltaromaticum, Candida zeylanoides (teleomorph: Kurtzmaniella zeylanoides), C. sake, and Debaryomyces hansenii. Two strains of C. zeylanoides and C. sake exhibiting high proteolytic and lipolytic activities effectively hydrolyzed beef fat, myofibrillar protein, and sarcoplasmic protein. This study characterized the main microorganisms and their enzymatic functions associated with dry-aged beef, highlighting the need to explore their contributions to the sensory attributes of dry-aged beef. Full article

The effect of Debaryomyces hansenii SH4, a typical aroma enhancer, on flavor formation of the dry fermented sausage was investigated using gas chromatography-mass spectrometry and metagenomic sequencing. The results showed that inoculation with D. hansenii SH4 promoted volatile compound formation from carbohydrate and amino acid metabolism and accelerated ester synthesis. The enzymes, genes, and microorganisms involved in the formation pathway of volatile compounds based on microbial metabolism were predicted and constructed into a metabolic pathway network. D. hansenii, Lactobacillus curvatus, Lactobacillus sakei, Lactobacillus plantarum, Leuconostoc fallax, Weissella minor, and Staphylococcus and Candida species were found to be the predominant functional microbes for flavor development in dry sausage. This study established a new insight into the metagenome-based bioinformatic effects of D. hansenii SH4 as a starter culture on the microbial synthesis of key volatile compounds in dry sausage. Full article

Efficient transcriptional regulation of the stress response is critical for microorganism survival. In yeast, stress-related gene expression, particularly for antioxidant enzymes like catalases, mitigates reactive oxygen species such as hydrogen peroxide (H₂O₂), preventing cell damage. The halotolerant yeast Debaryomyces hansenii shows oxidative stress tolerance, largely due to high catalase activity from DhCTA and DhCTT genes. This study evaluates D. hansenii’s response to oxidative stress caused by H₂O₂ under saline conditions, focusing on cell viability, gene expression, and catalase activity. Chromatin organization in the promoter of DhCTA and DhCTT was analyzed, revealing low nucleosome occupancy in promoter regions, correlating with active gene expression. Stress-related motifs for transcription factors like Msn2/4 and Sko1 were found, suggesting regulation by the DhHog1 MAP kinase. Analysis of a Dhhog1Δ mutant showed DhHog1’s role in DhCTA expression under H₂O₂ or NaCl conditions. These findings highlight DhHog1’s critical role in regulating the stress response in D. hansenii, offering insights for enhancing stress tolerance in halotolerant yeasts, particularly for industrial applications in saline wastewater management. Full article

The European “Green Deal” policies are shifting toward more sustainable and environmentally conscious agricultural practices, reducing the use of chemical fertilizer and pesticides. This implies exploring alternative strategies. One promising alternative to improve plant nutrition and reinforce plant defenses is the use of beneficial microorganisms in the rhizosphere, such as “Plant-growth-promoting rhizobacteria and fungi”. Despite the great abundance of iron (Fe) in the Earth’s crust, its poor solubility in calcareous soil makes Fe deficiency a major agricultural issue worldwide. Among plant promoting microorganisms, the yeast Debaryomyces hansenii has been very recently incorporated, for its ability to induce morphological and physiological key responses to Fe deficiency in plants, under hydroponic culture conditions. The present work takes it a step further and explores the potential of D. hansenii to improve plant nutrition and stimulate growth in cucumber plants grown in calcareous soil, where ferric chlorosis is common. Additionally, the study examines D. hansenii’s ability to induce systemic resistance (ISR) through a comparative relative expression study by qRT-PCR of ethylene (ET) biosynthesis (ACO1), or ET signaling (EIN2 and EIN3), and salicylic acid (SA) biosynthesis (PAL)-related genes. The results mark a significant milestone since D. hansenii not only enhances nutrient uptake and stimulates plant growth and flower development but could also amplify induced systemic resistance (ISR). Although there is still much work ahead, these findings make D. hansenii a promising candidate to be used for sustainable and environmentally friendly integrated crop management. Full article

The aim of this research was to determine the influence of yeast strains (previously isolated from the fermentation process) on selected quality parameters of sauerkraut. For this purpose, shredded and salted (2.5% w/w) cabbage of the Galaxy variety was fermented in the absence of oxygen with the addition of 2 × 10⁶ cells of a selected yeast culture. The control sample was spontaneously fermented sauerkraut without yeast addition. The obtained sauerkraut was analysed in terms of the content of selected organic acids, sugars and polyols (HPLC), selected volatile compounds (HS-SPME-GC-TOFMS), colour (CieLAB) and aroma (QDA). Yeast P. fermentans, Rh. mucilaginosa and W. anomalus reduced crucial sauerkraut components such as lactic acid, glycerol, and certain volatile compounds, leading to decreased aroma intensity and acceptability. Additionally, an increase in glucosinolate decomposition products was observed. Conversely, D. hansenii positively influenced sauerkraut quality by enhancing lactic acid content and exhibiting similar volatile characteristics to those of the control. Two of the three samples fermented with D. hansenii received high sensory analysis scores akin to those of the control. Sauerkraut fermented with Cl. lusitaniae yeast contained elevated levels of volatile compounds—alcohols, esters and lactones—resulting in an intense floral aroma, albeit receiving lower overall ratings due to deviation from the typical profile. Full article

In some countries, yeasts are still not allowed in the production of commercially fermented sausages. Therefore, further research is needed on producing fermented meat products using different strains of yeasts. In this study, two strains of Debaryomyces hansenii (D. hansenii Y61 and Y67) were inoculated in fermented sausages to study their effects as starter cultures. The inoculation of D. hansenii strains affected ripening by decreasing the pH and a_w. The sausages inoculated with Y61 and Y67 exhibited decreases in lipid oxidation of 40.70% and 36.04%, respectively, and Enterobacteriaceae counts of 50% and 100%, respectively. The inoculating yeasts Y61 and Y67 increased the lightness (L*) and redness (a*) of fermented sausages. The D. hansenii-inoculated sausages had higher levels of free amino acids and fatty acids, which improved the digestibility, sensory value, and safety of these sausages. Moreover, the total amount of ester compounds increased by 87.14% and 83.31% in the Y61- and Y67-inoculated groups, respectively, which contributed to the aroma. Better sensory attributes were also found in the sausages inoculated with Y61 and Y67 D. hansenii. Native D. hansenii Y61 and Y67 are, therefore, good starter cultures for fermented sausage production. Together, the results provide data supporting future research and the use of yeast-fermented sausages. Full article

The main objective of this work was to evaluate the combined effect of a biotechnology process, based on selected yeast strains, and a high-pressure homogenization (HPH) treatment on the microbiological quality, structural organization of proteins, chitin content, and antioxidant activity of a mixture of cricket powder (Acheta domesticus) and water. Compared to untreated samples, the cricket matrix treated with HPH four times at 180 MPa promoted the growth of the inoculated Yarrowia lipolytica and Debaryomyces hansenii strains. HPH did not affect the concentration of chitin; however, the combination with microorganisms tended to reduce the content. Although the antioxidant activity increased from 0.52 to 0.68 TAC mM/TE after a 48 h incubation in the control, it was further improved by the combination of HPH and D. hansenii metabolism, reaching a value of 0.77 TAC mM/TE. The combination of the two approaches also promoted a reduction in the intensity of bands with molecular weights between 31 and 21.5 kDa in favor of bands with a lower molecular weight. In addition, HPH treatment reduced the number of accessible thiols, suggesting protein structure changes that may further impact the technological properties of cricket powder. Full article

The extracted olive pomace (EOP) is an industrial lignocellulosic by-product of olive pomace oil extraction, currently mainly used for energy production through combustion. In this work, the hemicellulosic fraction of EOP was selectively hydrolyzed by diluted acid hydrolysis to obtain pentose-rich hydrolysates that can potentially be upgraded by Debaryomyces hansenii, targeting xylitol production. The monosaccharides and degradation by-products released along the pre-treatment were quantified and several detoxification methods for the removal of potentially toxic compounds were evaluated, including pH adjustment to 5.5, the use of anion-exchange resins, adsorption into activated charcoal, concentration by evaporation, and membrane techniques, i.e., nanofiltration. The latter approach was shown to be the best method allowing the full removal of furfural, 41% of 5-hydroxymethylfurfural, 54% of acetic acid, and 67% of the phenolic compounds present in the hydrolysate. The effects of the supplementation of both non-detoxified and detoxified hydrolysates were also assessed. The non-detoxified hydrolysate, under aerobic conditions, supported the yeast growth and xylitol production at low levels. Supplementation with the low-cost corn steep liquor of the nanofiltration detoxified hydrolysate showed a higher xylitol yield (0.57 g/g) compared to the non-detoxified hydrolysate. The highest xylitol productivity was found in hydrolysate detoxified with anionic resins (0.30 g/L·h), which was 80% higher than in the non-detoxified culture medium. Overall, the results showed that EOP dilute acid hydrolysates can efficiently be used for xylitol production by D. hansenii if detoxification, and supplementation, even with low-cost supplements, are performed. Full article

Debaryomyces hansenii is considered an unconventional yeast with a strong biotechnological potential, which can produce and store high amounts of lipids. However, relatively little is known about its lipid metabolism, and genetic tools for this yeast have been limited. The aim of this study was to explore the fatty acid β-oxidation pathway in D. hansenii. To this end, we employed recently developed methods to generate multiple gene deletions and tag open reading frames with GFP in their chromosomal context in this yeast. We found that, similar as in other yeasts, the β-oxidation of fatty acids in D. hansenii was restricted to peroxisomes. We report a series of experiments in D. hansenii and the well-studied yeast Saccharomyces cerevisiae that show that the homeostasis of NAD⁺ in D. hansenii peroxisomes is dependent upon the peroxisomal membrane protein Pmp47 and two peroxisomal dehydrogenases, Mdh3 and Gpd1, which both export reducing equivalents produced during β-oxidation to the cytosol. Pmp47 is the first identified NAD⁺ carrier in yeast peroxisomes. Full article

This study assessed the effect of replacing pork lard with coconut oil and Debaryomyces hansenii inoculation on the biotransformation of amino acids into volatile compounds in a meat model system. Yeast counts, solid-phase microextraction, and gas chromatography/mass spectrometry were used to assess yeast growth and volatile production, respectively. Yeast growth was confirmed until 28 d, although the volatile profile changed until 39 d. Forty-three volatiles were quantified, and their odor activity values (OAVs) were calculated. The presence of fat and yeasts contributed to differences in volatiles. In pork lard models, a delayed formation of lipid-derived aldehyde compounds was observed, whereas in coconut oil models, the generation of acid compounds and their respective esters was enhanced. Yeast activity affected amino acid degradation, which produced an increase in branched-chain aldehydes and alcohols. The aroma profile in the coconut models was influenced by hexanal, acid compounds, and their respective esters, whereas in pork lard models, aroma was affected by methional (musty, potato) and 3-methylbutanal (green, cocoa). The yeast inoculation contributed to the generation of 3-methylbutanoic acid (cheesy) and phenylethyl alcohol (floral). The type of fat and yeast inoculation produced a differential effect on the aroma. Full article