Spoilage Yeasts Research Articles

Spoilage yeasts in fermented vegetables: conventional and novel control strategies

Spoilage yeasts in fermented vegetables: conventional and novel control strategies

Zygosaccharomyces rouxii is the predominant species responsible for the spoilage of the mix base for ice cream and ethanol is the best inhibitor tested

A mix base for ice cream (MBIC) is used to produce artisanal or industrial ice creams and desserts and consists of a mixture of different ingredients, including sugar, egg yolk, natural flavors, starch and milk proteins. MBICs, which have chemical-physical characteristics that include a pH of 5.61 and an activity water (Aw) less than or equal to 0.822, are packaged in tin boxes and stored at ambient temperature. Despite the low Aw, MBIC can support osmotolerant and osmophilic yeast growth. The aim of our work was to study the behavior of Zygosaccharomyces rouxii, the main microorganisms responsible of MBIC spoilage, either in the vivo or in a model system in order to inhibit its growth by the selection of antimicrobial agents. Different osmotolerant yeasts belonging to the genus Zygosaccharomyces were isolated and identified from spoiled and unspoiled lots of MBICs. In particular, Z. rouxii was the predominant species responsible for the spoilage, which depended on the high temperature of storage (>20 °C) and was highlighted by the presence of alcohol, esters, acids and gas (CO2), which blew open the tin boxes. To stop spoilage, different antimicrobial compounds were tested: sulfur dioxide, sorbic and benzoic acids and ethanol. However, only 2% v/v ethanol was required to achieve the total inhibition of the Z. rouxii cocktails tested in this work. The use of other antimicrobials cannot be recommended because they were not able to stop yeast spoilage and changed the color and flavor of the products. Conversely, the use of ethanol is suggested because of its extreme effectiveness against osmotolerant yeasts, and the added amount was less than or equal to the taste threshold limit. The MBICs, treated with ethanol, were stable till the end of their shelf-life (6 months).

Antimycotic Effects of 11 Essential Oil Components and Their Combinations on 13 Food Spoilage Yeasts and Molds.

Food safety is important to reduce food spoilage microorganisms and foodborne pathogens. However, food safety is challenging, as customers’ demand for natural preservatives is increasing. Essential oils (EOs) and their components (EOCs) are alternative antibacterial and antimycotic food additives. In this study, the minimal inhibitory concentrations (MIC) of 11 different EOCs against 13 food spoilage molds and yeasts were investigated via the microdilution method. Cinnamaldehyde (CA) revealed the lowest MIC for all tested strains and all EOCs (32.81–328.1 µg ml−1). However, CA is organoleptic and was therefore combined with other EOCs via the checkerboard method. Overall, 27 out of 91 combinations showed a synergistic effect, and both respective EOC concentrations could be reduced by maintaining MIC. Thereby, the combination with citral or citronellal showed promising results. The concentration-dependent effect of CA was studied in further detail on Saccharomyces cerevisiae, with CA causing delayed growth-kinetics and reduced total cell numbers. In addition, flow cytometric measurements combined with live–dead staining indicate the fungicidal effect of CA, due to decreasing total cell numbers and increasing relative amount of propidium iodide-positive cells. In this study, we demonstrated that CA is a potent candidate for the use as a natural preservative against food-relevant mold and yeasts showing fungistatic and fungicidal effects. Therefore, CA and EOC combinations with respective lower EOC concentrations reduce organoleptic reservations, which ease their application in the food industry.

Emerging Non-Thermal Technologies as Alternative to SO2 for the Production of Wine.

SO2 is an antioxidant and selective antimicrobial additive, inhibiting the growth of molds in the must during the early stages of wine production, as well as undesirable bacteria and yeasts during fermentation, thus avoiding microbial spoilage during wine production and storage. The addition of SO2 is regulated to a maximum of 150–350 ppm, as this chemical preservative can cause adverse effects in consumers such as allergic reactions. Therefore, the wine industry is interested in finding alternative strategies to reduce SO2 levels, while maintaining wine quality. The use of non-thermal or cold pasteurization technologies for wine preservation was reviewed. The effect of pulsed electric fields (PEF), high pressure processing (HPP), power ultrasound (US), ultraviolet irradiation (UV), high pressure homogenization (HPH), filtration and low electric current (LEC) on wine quality and microbial inactivation was explored and the technologies were compared. PEF and HPP proved to be effective wine pasteurization technologies as they inactivate key wine spoilage yeasts, including Brettanomyces, and bacteria in short periods of time, while retaining the characteristic flavor and aroma of the wine produced. PEF is a promising technology for the beverage industry as it is a continuous process, requiring only microseconds of processing time for the inactivation of undesirable microbes in wines, with commercial scale, higher throughput production potential.

Effects of Gaseous Ozone on Microbiological Quality of Andean Blackberries (Rubus glaucus Benth).

Andean blackberries are highly perishable due to their susceptibility to water loss, softening, mechanical injuries, and postharvest diseases. In this study, the antimicrobial efficacy of gaseous ozone against spoilage (mesophiles, psychrotrophs, and yeasts and molds) and pathogenic (E. coli, S. enterica, and B. cinerea) microorganisms was evaluated during 10 days of storage at 6 ± 1 °C. Respiration rate and mass loss were also determined. Ozone was applied prior to storage at 0.4, 0.5, 0.6, and 0.7 ppm, for 3 min. The best results were observed with the higher ozone dose, with initial maximum reductions of ~0.5, 1.09, and 0.46 log units for E. coli, S. enterica, and B. cinerea, respectively. For the native microflora, maximum reductions of 1.85, 1.89, and 2.24 log units were achieved on day 1 for the mesophiles, psychrotrophs, and yeasts and molds, respectively, and this effect was maintained throughout storage. In addition, the lower respiration rate and mass loss of the blackberries ozonated at 0.7 ppm indicate that this treatment did not induce physiological damage to the fruit. Gaseous O3 could be effective in maintaining the postharvest quality of blackberries throughout refrigerated storage but higher doses could be advisable to enhance its antimicrobial activity.

Development of quantitative real-time PCR and digital droplet-PCR assays for rapid and early detection of the spoilage yeasts Saccharomycopsis fibuligera and Wickerhamomyces anomalus in bread

In the present study, for the first time, high sensitive quantitative polymerase chain reaction (qPCR) and digital droplet polymerase chain reaction (ddPCR) assays were developed to detect and quantify total eumycetes with potential application in several food matrices and to specifically determine the level of contamination by Saccharomycopsis fibuligera and Wickerhamomyces anomalus cells directly in bread. Among the candidate target genes used to develop the assays, car1 gene was chosen to detect the two spoilage yeasts S. fibuligera and W. anomalus. The specificity of the PCR assays was tested using purified genomic DNA from 36 bacterial and fungal strains. The sensitivity of the assays was defined by using tenfold serial dilutions of genomic DNA starting from 106 cfu/mL to 1 cfu/mL of S. fibuligera and W. anomalus. Validation of the assays was achieved by enumeration of S. fibuligera and W. anomalus DNA copies from samples of artificially contaminated bread homogenates detecting up to 10 cfu/mL (0.06 ± 0.01 copies/μL) of W. anomalus by using ddPCR. In conclusion, the developed qPCR and ddPCR assays demonstrate strong performance in the early detection of S. fibuligera and W. anomalus in bread, representing promising tools for applying high-throughput approaches to regularly monitor bread quality.

The N.C.Yeastract and CommunityYeastract databases to study gene and genomic transcription regulation in non-conventional yeasts.

Responding to the recent interest of the yeast research community in non-Saccharomyces cerevisiae species of biotechnological relevance, the N.C.Yeastract (http://yeastract-plus.org/ncyeastract/) was associated to YEASTRACT + (http://yeastract-plus.org/). The YEASTRACT + portal is a curated repository of known regulatory associations between transcription factors (TFs) and target genes in yeasts. N.C.Yeastract gathers all published regulatory associations and TF-binding sites for Komagataellaphaffii (formerly Pichia pastoris), the oleaginous yeast Yarrowia lipolytica, the lactose fermenting species Kluyveromyces lactis and Kluyveromyces marxianus, and the remarkably weak acid-tolerant food spoilage yeast Zygosaccharomyces bailii. The objective of this review paper is to advertise the update of the existing information since the release of N.C.Yeastract in 2019, and to raise awareness in the community about its potential to help the day-to-day work on these species, exploring all the information available in the global YEASTRACT + portal. Using simple and widely used examples, a guided exploitation is offered for several tools: (i) inference of orthologous genes; (ii) search for putative TF binding sites and (iii) inter-species comparison of transcription regulatory networks and prediction of TF-regulated networks based on documented regulatory associations available in YEASTRACT + for well-studied species. The usage potentialities of the new CommunityYeastract platform by the yeast community are also discussed.

Non-Saccharomyces as Biotools to Control the Production of Off-Flavors in Wines.

Off-flavors produced by undesirable microbial spoilage are a major concern in wineries, as they affect wine quality. This situation is worse in warm areas affected by global warming because of the resulting higher pHs in wines. Natural biotechnologies can aid in effectively controlling these processes, while reducing the use of chemical preservatives such as SO2. Bioacidification reduces the development of spoilage yeasts and bacteria, but also increases the amount of molecular SO2, which allows for lower total levels. The use of non-Saccharomyces yeasts, such as Lachancea thermotolerans, results in effective acidification through the production of lactic acid from sugars. Furthermore, high lactic acid contents (>4 g/L) inhibit lactic acid bacteria and have some effect on Brettanomyces. Additionally, the use of yeasts with hydroxycinnamate decarboxylase (HCDC) activity can be useful to promote the fermentative formation of stable vinylphenolic pyranoanthocyanins, reducing the amount of ethylphenol precursors. This biotechnology increases the amount of stable pigments and simultaneously prevents the formation of high contents of ethylphenols, even when the wine is contaminated by Brettanomyces.

Temporal Comparison of Microbial Community Structure in an Australian Winery

Most modern fermented foods and beverages are produced in fit-for-purpose facilities which are designed to ensure not only a reliable product, but also one safe for consumption. Despite careful hygiene, microorganisms can colonise these facilities and establish resident populations that can potentially contribute to the fermentation process. Although some microorganisms may not negatively affect the final product, spoilage microorganisms can be detrimental for quality, generating substantial economic losses. Here, amplicon-based phylotyping was used to map microbial communities within an Australian winery, before, during and after the 2020 vintage. Resident bacterial and yeast populations were shown to change over time, with both relative abundance and location within the winery varying according to sampling date. The bacterial family Micrococcaceae, and the genera Sphingomonas and Brevundimonas were the most abundant bacterial taxonomies, while Naganishia, Pyrenochaeta and Didymella were the most abundant fungal genera. Mapping the spatial distributions of the microbial populations identified the main locations that harboured these resident microorganisms, that include known wine spoilage yeasts and bacteria. Wine spoilage microorganisms, including the genefugura Lactobacillus, Acetobacter, Gluconobacter and Brettanomyces showed very low relative abundance and were found only in a couple of locations within the winery. Microbial populations detected in this facility were also compared to the resident microbiota identified in other fermented food facilities, revealing that microbial population structures may reflect the nature of the product created in each facility.

Prediction of Genetic Groups within Brettanomyces bruxellensis through Cell Morphology Using a Deep Learning Tool.

Brettanomyces bruxellensis is described as a wine spoilage yeast with many mainly strain-dependent genetic characteristics, bestowing tolerance against environmental stresses and persistence during the winemaking process. Thus, it is essential to discriminate B. bruxellensis isolates at the strain level in order to predict their stress resistance capacities. Few predictive tools are available to reveal intraspecific diversity within B. bruxellensis species; also, they require expertise and can be expensive. In this study, a Random Amplified Polymorphic DNA (RAPD) adapted PCR method was used with three different primers to discriminate 74 different B. bruxellensis isolates. High correlation between the results of this method using the primer OPA-09 and those of a previous microsatellite analysis was obtained, allowing us to cluster the isolates among four genetic groups more quickly and cheaply than microsatellite analysis. To make analysis even faster, we further investigated the correlation suggested in a previous study between genetic groups and cell polymorphism using the analysis of optical microscopy images via deep learning. A Convolutional Neural Network (CNN) was trained to predict the genetic group of B. bruxellensis isolates with 96.6% accuracy. These methods make intraspecific discrimination among B. bruxellensis species faster, simpler and less costly. These results open up very promising new perspectives in oenology for the study of microbial ecosystems.

In Vitro Activity of Selected Phenolic Compounds against Planktonic and Biofilm Cells of Food-Contaminating Yeasts.

Phenolic compounds are natural substances that can be obtained from plants. Many of them are potent growth inhibitors of foodborne pathogenic microorganisms, however, phenolic activities against spoilage yeasts are rarely studied. In this study, planktonic and biofilm growth, and the adhesion capacity of Pichia anomala, Saccharomyces cerevisiae, Schizosaccharomyces pombe and Debaryomyces hansenii spoilage yeasts were investigated in the presence of hydroxybenzoic acid, hydroxycinnamic acid, stilbene, flavonoid and phenolic aldehyde compounds. The results showed significant anti-yeast properties for many phenolics. Among the tested molecules, cinnamic acid and vanillin exhibited the highest antimicrobial activity with minimum inhibitory concentration (MIC) values from 500 µg/mL to 2 mg/mL. Quercetin, (−)-epicatechin, resveratrol, 4-hydroxybenzaldehyde, p-coumaric acid and ferulic acid were also efficient growth inhibitors for certain yeasts with a MIC of 2 mg/mL. The D. hansenii, P. anomala and S. pombe biofilms were the most sensitive to the phenolics, while the S. cerevisiae biofilm was quite resistant against the activity of the compounds. Fluorescence microscopy revealed disrupted biofilm matrix on glass surfaces in the presence of certain phenolics. Highest antiadhesion activity was registered for cinnamic acid with inhibition effects between 48% and 91%. The active phenolics can be natural interventions against food-contaminating yeasts in future preservative developments.

Development of a natamycin-based non-migratory antimicrobial active packaging for extending shelf-life of yogurt drink (Doogh)

A natamycin-based non-migratory antimicrobial packaging for extending shelf-life of yogurt drink (Doogh) was developed. Firstly, the surface of low-density polyethylene film (LDPE) was modified with acrylic acid at different times of UV exposure (0–10 min) to produce carboxylic functional groups. Then, natamycin was applied to the UV-treated films to bind covalently with the pendent functional groups. The maximum grafting efficiency (81.96%) was obtained for the 6 min treated film. Moreover, surface properties of films were evaluated by Attenuated Total Reflectance/Fourier Transfer Infrared Spectroscopy (ATR-FTIR) and scanning electron microscopy (SEM). Antifungal activity of different treatments of natamycin grafted film was evaluated against two common spoilage yeasts of Doogh including Rhodotorula mucilaginosa and Candida parapsilosis. Results showed that 6 min treated film provides maximum anti-yeast activity and can be applied to control fungal growth in Doogh. Natamycin-grafted film postponed the yeast spoilage in Doogh and prolonged its shelf-life to 23 days.

Wine yeast species show strong inter- and intra-specific variability in their sensitivity to ultraviolet radiation

While the trend in winemaking is toward reducing the inputs and especially sulphites utilization, emerging technologies for the preservation of wine is a relevant topic for the industry. Amongst yeast spoilage in wine, Brettanomyces bruxellensis is undoubtedly the most feared. In this study, UV-C treatment is investigated. This non-thermal technique is widely used for food preservation. A first approach was conducted using a drop-platted system to compare the sensitivity of various strains to UV-C surface treatment. 147 strains distributed amongst fourteen yeast species related to wine environment were assessed for six UV-C doses. An important variability in UV-C response was observed at the interspecific level. Interestingly, cellar resident species, which are mainly associated with wine spoilage, shows higher sensitivity to UV-C than vineyard-resident species. A focus on B. bruxellensis species with 104 screened strains highlighted an important effect of the UV-C, with intra-specific variation. This intra-specific variation was confirmed on 6 strains in liquid red wine by using a home-made pilot. 6624 J.L−1 was enough for a reduction of 5 log10 of magnitude for 5 upon 6 strains. These results highlight the potential of UV-C utilization against wine yeast spoiler at cellar scale.

Development of predictive models evaluating the spoilage-delaying effect of a bioprotective culture on different yeast species in yogurt

Yeast spoilage of fermented dairy products causes challenges for the dairy industry, including economic losses due to wasted product. Food cultures with bioprotective effects are becoming more widely used to help ensure product quality throughout product shelf life. To assist the dairy industry when evaluating product quality throughout shelf life and the effect of bioprotective cultures, we aimed to build stochastic models that provide reliable predictions of yeast spoilage in yogurt with and without bioprotective culture. Growth characterizations of Debaryomyces hansenii, Yarrowia lipolytica, Saccharomyces cerevisiae, and Kluyveromyces marxianus at storage temperatures of 7, 12, and 16°C during a 30-d storage period were conducted in yogurt with and without a bioprotective culture containing Lacticaseibacillus rhamnosus strains. The kinetic growth parameters were calculated using the Buchanan growth model, and these parameters were used as baseline values in Monte Carlo models to translate the yeast growth into spoilage levels. The models were developed using 100,000 simulations and they predicted yeast spoilage levels in yogurt by the 4 yeast types. Each modeled yogurt batch was set to be contaminated with yeast at a concentration drawn from a normal distribution with a mean of 1 log10 cfu/mL and standard deviation of 1 log10 cfu/mL and stored for 30 d at a temperature drawn from a normal distribution with a mean of 6.1°C and a standard deviation of 2.8°C. Considering a spoilage level of 5 log10 cfu/mL, the predicted number of spoiled samples was reduced 3-fold during the first 10 d and by 2-fold at the end of shelf life when a bioprotective culture was added to the yogurt. The models were evaluated by sensitivity analyses, where the main effect factors were maximum yeast population, storage temperature, and yeast strain. The models were validated by comparing the model output to actual observed spoilage data from a European dairy using the bioprotective culture. When the model prediction, based on a mixture of the 4 specific yeast strains, was compared with spoilage data from the European dairy, the observed effect of bioprotective cultures was considerably higher than predicted, potentially influenced by the presence of contaminating strains more sensitive to a bioprotective culture than those characterized here. The developed Monte Carlo models can predict yeast spoilage levels in yogurt at specific production settings and how this may be affected by various parameters and addition of bioprotective cultures.

Potential application of ginger, clove and thyme essential oils to improve soft cheese microbial safety and sensory characteristics

Due to consumers growing concerns about synthetic preservatives, essential oils (EOs) come into the focus of pharmaceutical and food applications as natural alternatives because of their inherent antimicrobial activity. However, EOs food applications have been challenged by their strong aroma that adversely affects consumers acceptability. In this study, EOs of Zingiber officinale (ginger), Syzygium aromaticum (clove), and Thymus vulgaris (thyme) were extracted and assessed for their potential use to produce cheese with high degree of microbial safety and sensory acceptability. Phytochemical profiling showed camphene and zingiberene as major constituents in ginger oil, whereas thymol and eugenol represented the chief volatiles in thyme and clove, respectively. The EOs antimicrobial activity against seven strains of pathogenic and spoilage bacteria, yeast and mold revealed MIC of 0.001%. Additionally, studied EOs demonstrated interesting safety profile displaying good to very weak cytotoxicity on human hepatocellular carcinoma (HepG2) with CC50 6.31–452.95 μg/mL and very low cytotoxicity on normal human lung fibroblasts (WI-38) with CC50 728.94–841.66 μg/mL. Sensory characterization of EOs-fortified laboratory manufactured fresh soft cheese displayed scores significantly higher than control samples (P < 0.001) in relation to overall acceptability. Viability study of Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa in fortified cheese matrix showed complete reduction of S. aureus at the end of first and second weeks of storage of thyme and ginger fortified cheese, respectively, and about 50% reduction of P. aeruginosa by the end of storage period. This study opens interesting new perspectives for the agro-industrial applications of EOs as natural preservatives.

Stability and Activity of the Antimicrobial Peptide Leg1 in Solution and on Meat and Its Optimized Generation from Chickpea Storage Protein.

The antimicrobial peptide Leg1 (RIKTVTSFDLPALRFLKL) from chickpea legumin is active against spoilage bacteria, yeast, and mold. The present study tested its effectiveness under food storage conditions and examined options to obtain a food-grade agent. The minimum inhibitory concentration (MIC) of Leg1 against E. coli (62.5 µM) proved stable over seven days at 20 °C or 4 °C. It was not influenced by reduced pH (5.0 vs. 6.8), which is relevant in food such as meat. An incubation temperature of 20 °C vs. 37 °C reduced the MIC to 15.6/7.8 µM against E. coli/B. subtilis. With a minimum bactericidal concentration in meat of 125/15.6 µM against E. coli/B. subtilis, Leg1 is equivalently effective as nisin and 5000–82,000 times more active than sodium benzoate, potassium sorbate, or sodium nitrite. Replacing the counter-ion trifluoroacetate derived from peptide synthesis by the more natural alternatives acetate or chloride did not impair the activity of Leg1. As an alternative to chemical synthesis, an optimized protocol for chymotryptic hydrolysis was developed, increasing the yield from chickpea legumin by a factor of 30 compared to the standard procedure. The present results indicate that food-grade Leg1 could possibly be applicable for food preservation.

Toward application of biocontrol to inhibit wine spoilage yeasts: The use of statistical designs for screening and optimisation

Spoilage yeasts generate considerable economic losses in the wine industry, and although sulphur dioxide (SO2) is traditionally used for control, its use has become controversial because of its negative effects on health. Biocontrol has emerged as a partial alternative to SO2, and most research has focused on the selection of biocontrol yeasts and/or the mechanisms involved, while little research has been directed to the environmental conditions that make biocontrol effective for application. When there are two or more interacting yeasts, the physicochemical factors that affect their antagonism are many and therefore the application of biocontrol is complex. To reduce SO2, the present study aimed to elucidate biocontrol mechanisms of two yeast interactions and to establish optimal physicochemical conditions for biocontrol of the spoilage yeast during grape must fermentation. Through the use of statistical design, it was possible to find relevant physicochemical factors and optimise them. Wickerhamomyces anomalus “BWa156” developed an active supernatant against ZygoSaccharomyces rouxii “BZr6” while supernatant from Metschnikowia pulcherrima “BMp29” was ineffective. In mixed must fermentations, the first interaction (BWa156 vs. BZr6) showed fewer physicochemical factors impacting biocontrol compared to the second interaction (BMp29 vs. BZr6). However, the fewer factors of the first interaction had a stronger effect on the decline in the spoilage population. Validations showed that the optimal conditions for biocontrol with the first interaction could be predicted. Analysis of the results with BWa156 vs. BZr6 and BMp29 vs. BZr6 suggests that the first interaction is a competition that includes a killer toxin, while the second interaction involves competition for iron resources. Response surface methodology (RSM) allowed a reduction in the number of experiments and permitted to find the optimal biocontrol conditions (SO2: 0 mg mL-1; pH: 3.7; Reducing sugars: 23 °Brix) for the interaction between BWa156 and BZr6.

Growth Prediction of the Food Spoilage Yeast Debaryomyces Hansenii using Multivariate Data Analysis

The main aim of the present study was to predict the growth of the food spoilage yeast Debaryomyces hansenii by multivariate data analysis (MVDA) using temperature, pH and NaCl concentration as growth parameters. Growth of five strains of D. hansenii (DHI, DHII, DHIII, DHIV and DHV) was measured as optical density at 620 nm (OD620) at different values of temperature, pH and NaCl concentrations. It was found that salt was the most important factor, which affects yeast growth followed by temperature. The growth of all yeast strains was reduced by increasing salt concentration and decreasing temperature. On the other hand, pH was found to have a little effect on the growth of D. hansenii. Strain DHII was the most salt-tolerant strains among the five yeast strains investigated. Partial least squares (PLS) prediction model was created out using pH, temperature and NaCl concentration to predict the growth of D. hansenii. The model was acceptable with a correlation of 0.86. The developed PLS model will help in optimizing the food process conditions that will prevent food spoilage by D. hansenii.

Growth Prediction of the Food Spoilage Yeast Debaryomyces Hansenii using Multivariate Data Analysis

The main aim of the present study was to predict the growth of the food spoilage yeast Debaryomyces hansenii by multivariate data analysis (MVDA) using temperature, pH and NaCl concentration as growth parameters. Growth of five strains of D. hansenii (DHI, DHII, DHIII, DHIV and DHV) was measured as optical density at 620 nm (OD620) at different values of temperature, pH and NaCl concentrations. It was found that salt was the most important factor, which affects yeast growth followed by temperature. The growth of all yeast strains was reduced by increasing salt concentration and decreasing temperature. On the other hand, pH was found to have a little effect on the growth of D. hansenii. Strain DHII was the most salt-tolerant strains among the five yeast strains investigated. Partial least squares (PLS) prediction model was created out using pH, temperature and NaCl concentration to predict the growth of D. hansenii. The model was acceptable with a correlation of 0.86. The developed PLS model will help in optimizing the food process conditions that will prevent food spoilage by D. hansenii.

Growth Inhibitory Effect of Garlic Powder and Cinnamon Extract on White Colony-Forming Yeast in Kimchi.

White colony-forming yeast (WCFY), also referred to as film forming yeast or spoilage yeast, that appear on the surface of kimchi can deteriorate the sensory properties of kimchi, such as odor and texture. Thus, the aim of this study was to develop a method to inhibit the formation of the white colony in kimchi. First, alterations in kimchi manufacturing and storage conditions, including temperatures, pH, salinity, and anaerobic condition, were investigated to determine if they could inhibit the growth of WCFY (i.e., Kazachstania servazzii, Candida sake, Debaryomyces hansenii, Pichia kudriavzevii, and Hanseniaspora uvarum). Thereafter, the anti yeast activity of freeze-dried garlic powder (FGP) and cinnamon ethanol extract (CEE) was evaluated against WCFY using the agar-well diffusion assay. Following the direct application of FGP and CEE to the surface of the kimchi, the inhibitory effects on white colony were determined. The results showed that WCFY can grow under various manufacturing and storage conditions of kimchi. Regarding the growth inhibitory effect on WCFY, FGP exhibited anti yeast activity against four WCFYs. It did not show anti yeast activity against K. servazzii. However, CEE showed anti yeast activity against K. servazzii. In particular, the mixture of 10% FGP and 1.75% CEE, which was manufactured considering the influence of sensory properties in kimchi, exhibited anti yeast activity against all WCFY. Furthermore, the application of the FGP and CEE mixture supplemented with 0.02% xanthan gum to kimchi to enhance adhesion to the kimchi surface, led to a delay in the formation of a white colony on the surface of the kimchi by an average of 17 d at 10 °C compared to the control group. Collectively, the use of a FGP, CEE, and xanthan gum mixture could be an effective method for the inhibition of white colony formation on the surface of kimchi, extending its shelf life.