scholarly journals Purification and Characterization of WA18, a New Mycocin Produced by Wickerhamomyces anomalus Active in Wine Against Brettanomyces bruxellensis Spoilage Yeasts

2020 ◽  
Vol 9 (1) ◽  
pp. 56
Author(s):  
Francesca Comitini ◽  
Alice Agarbati ◽  
Laura Canonico ◽  
Edoardo Galli ◽  
Maurizio Ciani
Keyword(s):  
Competitive Inhibition ◽  
Receptor Site ◽  
Restriction Enzymes ◽  
Culture Conditions ◽  
Starter Cultures ◽  
Cell Wall Polysaccharides ◽  
Brettanomyces Bruxellensis ◽  
Killer Activity ◽  
Wickerhamomyces Anomalus ◽  
Spoilage Yeasts

Wickerhamomyces anomalus strain 18, isolated from a natural underground cheese ripening pit, secretes a mycocin named WA18 that inhibits wine spoilage yeasts belonging to Brettanomyces bruxellensis species, with a broad-spectrum of activity. WA18 was purified, and the purified protein was digested with specific restriction enzymes (lysine K and arginine R cut sites). The LC–MS and LC–MS/MS analysis after enzymatic digestions revealed a molecular weight of 31 kDa. Bioinformatics processing and database research of digested pure killer protein showed 99% identity with a UDP-glycosyltransferase protein. Competitive inhibition assay of killer activity by cell-wall polysaccharides suggests that branched glucans represent the first receptor site of the toxin on the envelope of the sensitive target. The WA18 partially purified crude extract (PPCE) showed high stability of antimicrobial activity at the physicochemical conditions suitable for the winemaking process. Indeed, in wine WA18 was able to counteract B. bruxellensis and control the production of ethyl phenols. In addition, the strain WA18 was compatible with Saccharomyces cerevisiae in co-culture conditions with a potential application together with commercial starter cultures. These data suggest that WA18 mycocin is a promising biocontrol agent against spoilage yeasts in winemaking, particularly during wine storage.

Kluyveromyces phaffii killer toxin active against wine spoilage yeasts: purification and characterization

Microbiology ◽  
2004 ◽  
Vol 150 (8) ◽  
pp. 2535-2541 ◽  
Author(s):  
Francesca Comitini ◽  
Natalia Di Pietro ◽  
Laura Zacchi ◽  
Ilaria Mannazzu ◽  
Maurizio Ciani
Keyword(s):  
Biochemical Characterization ◽  
Competitive Inhibition ◽  
Killer Toxin ◽  
Receptor Site ◽  
Wine Industry ◽  
Cell Wall Polysaccharides ◽  
Killer Activity ◽  
Spoilage Yeast ◽  
Spoilage Yeasts ◽  
Potential Applications

The killer toxin secreted by Kluyveromyces phaffii (KpKt) is active against spoilage yeast under winemaking conditions and thus has potential applications in the biocontrol of undesired micro-organisms in the wine industry. Biochemical characterization and N-terminal sequencing of the purified toxin show that KpKt is a glycosylated protein with a molecular mass of 33 kDa. Moreover, it shows 93 % and 80 % identity to a β-1,3-glucanase of Saccharomyces cerevisiae and a β-1,3-glucan transferase of Candida albicans, respectively, and it is active on laminarin and glucan, thus showing a β-glucanase activity. Competitive inhibition of killer activity by cell-wall polysaccharides suggests that glucan (β-1,3 and β-1,6 branched glucans) represents the first receptor site of the toxin on the envelope of the sensitive target. Flow cytometry analysis of the sensitive target after treatment with KpKt and K1 toxin of S. cerevisiae, known to cause loss of cell viability via formation of pores in the cell membrane, suggests a different mode of action for KpKt.

scholarly journals PMKT2, a new killer toxin from Pichia membranifaciens, and its promising biotechnological properties for control of the spoilage yeast Brettanomyces bruxellensis

Microbiology ◽  
2009 ◽  
Vol 155 (2) ◽  
pp. 624-634 ◽  
Author(s):  
A. Santos ◽  
M. San Mauro ◽  
E. Bravo ◽  
D. Marquina
Keyword(s):  
Biochemical Characterization ◽  
Gel Filtration ◽  
Chemical Properties ◽  
Killer Toxin ◽  
Small Scale ◽  
Brettanomyces Bruxellensis ◽  
Killer Activity ◽  
Pichia Membranifaciens ◽  
Spoilage Yeast ◽  
Spoilage Yeasts

Pichia membranifaciens CYC 1086 secretes a killer toxin (PMKT2) that is inhibitory to a variety of spoilage yeasts and fungi of agronomical interest. The killer toxin in the culture supernatant was concentrated by ultrafiltration and purified to homogeneity by two successive steps, including native electrophoresis and HPLC gel filtration. Biochemical characterization of the toxin showed it to be a protein with an apparent molecular mass of 30 kDa and an isoelectric point of 3.7. At pH 4.5, optimal killer activity was observed at temperatures up to 20 °C. Above approximately this pH, activity decreased sharply and was barely noticeable at pH 6. The toxin concentrations present in the supernatant during optimal production conditions exerted a fungicidal effect on a variety of fungal and yeast strains. The results obtained suggest that PMKT2 has different physico-chemical properties from PMKT as well as different potential uses in the biocontrol of spoilage yeasts. PMKT2 was able to inhibit Brettanomyces bruxellensis while Saccharomyces cerevisiae was fully resistant, indicating that PMKT2 could be used in wine fermentations to avoid the development of the spoilage yeast without deleterious effects on the fermentative strain. In small-scale fermentations, PMKT2, as well as P. membranifaciens CYC 1086, was able to inhibit B. bruxellensis, verifying the biocontrol activity of PMKT2 in simulated winemaking conditions.

Optimization of Culture Conditions for Growth Associated with Cr(VI) Removal by Wickerhamomyces anomalus M10

2016 ◽  
Vol 98 (3) ◽  
pp. 400-406 ◽  
Author(s):  
Pablo Marcelo Fernández ◽  
Elías Leonardo Cruz ◽  
Silvana Carolina Viñarta ◽  
Lucía Inés Castellanos de Figueroa
Keyword(s):  
Culture Conditions ◽  
Wickerhamomyces Anomalus ◽  
Optimization Of Culture Conditions

scholarly journals Dynamic modelling of the killing mechanism of action by virus-infected yeasts

2019 ◽  
Vol 16 (152) ◽  
pp. 20190064
Author(s):  
Sean Sheppard ◽  
Duygu Dikicioglu
Keyword(s):  
Large Scale ◽  
Mechanistic Model ◽  
Dynamic Modelling ◽  
Starter Cultures ◽  
Apoptotic Pathway ◽  
Toxic Activity ◽  
Killer Activity ◽  
Killer Yeasts ◽  
Death Kinetics ◽  
Kinetics Of

Killer yeasts are microorganisms, which can produce and secrete proteinaceous toxins, a characteristic gained via infection by a virus. These toxins are able to kill sensitive cells of the same or a related species. From a biotechnological perspective, killer yeasts are beneficial due to their antifungal/antimicrobial activity, but also regarded as problematic for large-scale fermentation processes, whereby those yeasts would kill starter cultures species and lead to stuck fermentations. Here, we propose a mechanistic model of the toxin-binding kinetics pertaining to the killer population coupled with the toxin-induced death kinetics of the sensitive population to study toxic action. The dynamic model captured the transient toxic activity starting from the introduction of killer cells into the culture at the time of inoculation through to induced cell death. The kinetics of K1/K2 activity via its primary pathway of toxicity was 5.5 times faster than its activity at low concentration inducing the apoptotic pathway in sensitive cells. Conversely, we showed that the primary pathway for K28 was approximately three times slower than its equivalent apoptotic pathway, indicating the particular relevance of K28 in biotechnological applications where the toxin concentration is rarely above those limits to trigger the primary pathway of killer activity.

scholarly journals Adhesion of Dextran to Streptococcus muttans

Microbiology ◽  
2000 ◽  
Vol 81 (2) ◽  
pp. 485-489 ◽  
Author(s):  
J. KELSTRUP ◽  
T. D. FUNDER-NIELSEN
Keyword(s):  
Streptococcus Mutans ◽  
Sodium Dodecyl Sulphate ◽  
Competitive Inhibition ◽  
Sodium Dodecyl ◽  
Periodate Oxidation ◽  
Receptor Site ◽  
Spatial Arrangement ◽  
Borohydride Reduction ◽  
Reactive Site ◽  
Sugar Alcohols

Summary: Fifteen strains of Streptococcus mutans were agglutinated by 5 to 50 ng Leuco-nostoc dextran/ml, and one by 500 ng/ml. Competitive inhibition of the agglutination by sugars or sugar alcohols suggested that the spatial arrangement of the HOC(3)H and O(1) groups were important in the dextran but that the dextran receptor is not identical with the dextran reactive site of glucosyl transferase. Periodate oxidation of the dextran, with or without subsequent borohydride reduction, prevented agglutination, again implicating the C3 group. Pretreatment of dextran with cyclohexyl-isocyanide or of S. mutans with 4.0 m-urea, 0.01 m-EDTA or 0.1 % sodium dodecyl sulphate prevented agglutination; the last three reagents dispersed organisms already agglutinated. Ca2+ or Mg2+ reversed the effect of EDTA. Exposure of bacteria to 60°C or to papain for a few minutes impaired agglutination, so the dextran receptor site may be protein.

scholarly journals Effect of Candida intermedia LAMAP1790 Antimicrobial Peptides against Wine-Spoilage Yeasts Brettanomyces bruxellensis and Pichia guilliermondii

Fermentation ◽  
2020 ◽  
Vol 6 (3) ◽  
pp. 65 ◽  
Author(s):  
Rubén Peña ◽  
Jeniffer Vílches ◽  
Camila G.-Poblete ◽  
María Angélica Ganga
Keyword(s):  
Antimicrobial Peptides ◽  
Membrane Damage ◽  
Pichia Guilliermondii ◽  
Yeast Saccharomyces Cerevisiae ◽  
Brettanomyces Bruxellensis ◽  
Candida Intermedia ◽  
Spoilage Yeast ◽  
Spoilage Yeasts ◽  
Wine Spoilage ◽  
Fungicide Activity

Wine spoilage yeasts are one of the main issues in the winemaking industry, and the control of the Brettanomyces and Pichia genus is an important goal to reduce economic loses from undesired aromatic profiles. Previous studies have demonstrated that Candida intermedia LAMAP1790 produces antimicrobial peptides of molecular mass under 10 kDa with fungicide activity against Brettanomyces bruxellensis, without affecting the yeast Saccharomyces cerevisiae. So far, it has not been determined whether these peptides show biocontroller effect in this yeast or other spoilage yeasts, such as Pichia guilliermondii. In this work, we determined that the exposure of B. bruxellensis to the low-mass peptides contained in the culture supernatant of C. intermedia LAMAP1790 produces a continuous rise of reactive oxygen species (ROS) in this yeast, without presenting a significant effect on membrane damage. These observations can give an approach to the antifungal mechanism. In addition, we described a fungicide activity of these peptides fraction against two strains of P. guilliermondii in a laboratory medium. However, carrying out assays on synthetic must, peptides must show an effect on the growth of B. bruxellensis. Moreover, these results can be considered as a start to develop new strategies for the biocontrol of spoilage yeast.

scholarly journals Raman Spectroscopy and Chemometrics for Identification and Strain Discrimination of the Wine Spoilage Yeasts Saccharomyces cerevisiae, Zygosaccharomyces bailii, and Brettanomyces bruxellensis

2013 ◽  
Vol 79 (20) ◽  
pp. 6264-6270 ◽  
Author(s):  
Susan B. Rodriguez ◽  
Mark A. Thornton ◽  
Roy J. Thornton
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Raman Spectroscopy ◽  
High Sensitivity ◽  
Brettanomyces Bruxellensis ◽  
Content Type ◽  
Zygosaccharomyces Bailii ◽  
Spoilage Yeasts ◽  
Wine Spoilage ◽  
Novel Method ◽  
Classification Tool

ABSTRACTThe yeastsZygosaccharomyces bailii,Dekkera bruxellensis(anamorph,Brettanomyces bruxellensis), andSaccharomyces cerevisiaeare the major spoilage agents of finished wine. A novel method using Raman spectroscopy in combination with a chemometric classification tool has been developed for the identification of these yeast species and for strain discrimination of these yeasts. Raman spectra were collected for six strains of each of the yeastsZ. bailii,B. bruxellensis, andS. cerevisiae. The yeasts were classified with high sensitivity at the species level: 93.8% forZ. bailii, 92.3% forB. bruxellensis, and 98.6% forS. cerevisiae. Furthermore, we have demonstrated that it is possible to discriminate between strains of these species. These yeasts were classified at the strain level with an overall accuracy of 81.8%.

Modeling the Efficacy of Triplet Antimicrobial Combinations: Yeast Suppression by Lauric Arginate, Cinnamic Acid, and Sodium Benzoate or Potassium Sorbate as a Case Study

2010 ◽  
Vol 73 (3) ◽  
pp. 515-523 ◽  
Author(s):  
YUMEI DAI ◽  
MARK D. NORMAND ◽  
JOCHEN WEISS ◽  
MICHA PELEG
Keyword(s):  
Optical Density ◽  
Cinnamic Acid ◽  
Sodium Benzoate ◽  
Three Dimensional ◽  
Potassium Sorbate ◽  
Brettanomyces Bruxellensis ◽  
Yeast Cultures ◽  
Spoilage Yeasts ◽  
Lauric Arginate ◽  
Equal Efficacy

The growth of four spoilage yeasts, Saccharomyces cerevisiae, Zygosaccharomyces bailii, Brettanomyces bruxellensis, and Brettanomyces naardenensis, was inhibited with three-agent (triplet) combinations of lauric arginate, cinnamic acid, and sodium benzoate or potassium sorbate. The inhibition efficacy was determined by monitoring the optical density of yeast cultures grown in microtiter plates for 7 days. The relationship between the optical density and the sodium benzoate and potassium sorbate concentrations followed a single-term exponential decay model. The critical effective concentration was defined as the concentration at which the optical density was 0.05, which became an efficacy criterion for the mixtures. Critical concentrations of sodium benzoate or potassium sorbate as a function of the lauric arginate and cinnamic acid concentrations were then fitted with an empirical model that mapped three-agent combinations of equal efficacy. The contours of this function are presented in tabulated form and as two- and three-dimensional plots. Triplet combinations were highly effective against all four spoilage yeasts at three practical pH levels, especially at pH 3.0. The triplet combinations were particularly effective for inhibiting growth of Z. bailii, and combinations containing potassium sorbate had synergistic activities. The equal efficacy concentration model also allowed tabulation of the cost of the various combinations of agents and identification of those most economically feasible.

Sign in / Sign up

Export Citation Format

Share Document