Effects of Pichia kluyveri killer toxin on sensitive cells

1980 ◽  
Vol 46 (2) ◽  
pp. 205-220 ◽  
Author(s):  
E. J. Middelbeek ◽  
C. Stumm ◽  
G. D. Vogels
Keyword(s):  
Killer Toxin ◽  
Pichia Kluyveri

The incidence of killer activity and extracellular proteases in tropical yeast communities

1997 ◽  
Vol 43 (4) ◽  
pp. 328-336 ◽  
Author(s):  
Jacqueline Abranches ◽  
Leda C. Mendonça-Hagler ◽  
Allen N. Hagler ◽  
Paula B. Morais ◽  
Carlos A. Rosa
Keyword(s):  
Killer Toxin ◽  
Spatial Separation ◽  
Extracellular Proteases ◽  
Temporal Separation ◽  
Killer Activity ◽  
Killer Yeasts ◽  
Sporobolomyces Roseus ◽  
Nitrogen Nutrients ◽  
Pichia Kluyveri ◽  
Ascomycetous Yeasts

The presence of killer and proteolytic yeasts was studied among 944 isolates representing 105 species from tropical yeast communities. We found 13 killer toxin producing species, with Pichia kluyveri being the most frequent. Other killer yeast isolates were Candida apis, Candida bombicola, Candida fructus, Candida krusei, Candida sorbosa, Hanseniaspora uvarum, Issatchenkia occidentalis, Kloeckera apis, Kluyveromyces marxianus, Pichia membranaefaciens, Pichia ohmeri-like, and Sporobolomyces roseus. The communities from which killer yeasts were isolated had strains sensitive to them, and there were interspecific and intraspecific differences in the spectra of their killer activities. Pichia kluyveri had the broadest spectra of activity against sensitive isolates, and it apparently produced different toxins. The coexistence of sensitive and killer yeasts using the same substrate suggests that there is spatial separation in microhabitats or temporal separation in different stages of successions. Basidiomycetous yeasts were more frequently proteolytic than ascomycetous yeasts. Extracellular proteases could be important for the yeasts to have access to more nitrogen nutrients and obtain a better balance with available carbon sources.Key words: killer yeasts, extracellular proteases, tropical yeast communities.

Killer toxin producing strains of the yeasts Hanseniaspora uvarum and Pichia kluyveri

1988 ◽  
Vol 149 (3) ◽  
pp. 261-267 ◽  
Author(s):  
J. Zorg ◽  
S. Kilian ◽  
F. Radler
Keyword(s):  
Killer Toxin ◽  
Hanseniaspora Uvarum ◽  
Pichia Kluyveri

The ecological role of killer yeasts in natural communities of yeasts

1987 ◽  
Vol 33 (9) ◽  
pp. 783-796 ◽  
Author(s):  
William T. Starmer ◽  
Philip F. Ganter ◽  
Virginia Aberdeen ◽  
Marc-Andre Lachance ◽  
Herman J. Phaff
Keyword(s):  
Yeast Species ◽  
Kluyveromyces Lactis ◽  
Killer Toxin ◽  
Toxin Production ◽  
Killer Activity ◽  
Killer Yeast ◽  
Killer Toxins ◽  
Naturally Occurring ◽  
Pichia Kluyveri

The killer phenomenon of yeasts was investigated in naturally occurring yeast communities. Yeast species from communities associated with the decaying stems and fruits of cactus and the slime fluxes of trees were studied for production of killer toxins and sensitivity to killer toxins produced by other yeasts. Yeasts found in decaying fruits showed the highest incidence of killing activity (30/112), while yeasts isolated from cactus necroses and tree fluxes showed lower activity (70/699 and 11/140, respectively). Cross-reaction studies indicated that few killer-sensitive interactions occur within the same habitat at a particular time and locality, but that killer-sensitive reactions occur more frequently among yeasts from different localities and habitats. The conditions that should be optimal for killer activity were found in fruits and young rots of Opuntia cladodes where the pH is low. The fruit habitat appears to favor the establishment of killer species. Killer toxin may affect the natural distribution of the killer yeast Pichia kluyveri and the sensitive yeast Cryptococcus cereanus. Their distributions indicate that the toxin produced by P. kluyveri limits the occurrence of Cr. cereanus in fruit and Opuntia pads. In general most communities have only one killer species. Sensitive strains are more widespread than killer strains and few species appear to be immune to all toxins. Genetic study of the killer yeast P. kluyveri indicates that the mode of inheritance of killer toxin production is nuclear and not cytoplasmic as is found in Saccharomyces cerevisiae and Kluyveromyces lactis.

Production, purification and properties of a Pichia kluyveri killer toxin

1979 ◽  
Vol 45 (3) ◽  
pp. 437-450 ◽  
Author(s):  
E. J. Middelbeek ◽  
J. M. H. Hermans ◽  
C. Stumm
Keyword(s):  
Killer Toxin ◽  
Purification And Properties ◽  
Pichia Kluyveri

Physiological conditions affecting the sensitivity of Saccharomyces cerevisiae to a Pichia kluyveri killer toxin and energy requirement for toxin action

1980 ◽  
Vol 46 (5) ◽  
pp. 483-497 ◽  
Author(s):  
E. J. Middelbeek ◽  
H. H. A. M. van de Laar ◽  
J. M. H. Hermans ◽  
C. Stumm ◽  
G. D. Vogels
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Energy Requirement ◽  
Killer Toxin ◽  
Physiological Conditions ◽  
Pichia Kluyveri

scholarly journals High Potential of Pichia kluyveri and Other Pichia Species in Wine Technology

2021 ◽  
Vol 22 (3) ◽  
pp. 1196
Author(s):  
Javier Vicente ◽  
Fernando Calderón ◽  
Antonio Santos ◽  
Domingo Marquina ◽  
Santiago Benito
Keyword(s):  
Ethanol Yield ◽  
Color Stability ◽  
Quality Parameters ◽  
Wine Industry ◽  
Wine Aroma ◽  
Wine Quality ◽  
Promising Tool ◽  
Pichia Kluyveri ◽  
The Impact ◽  
Fermentative Yeast

The surfaces of grapes are covered by different yeast species that are important in the first stages of the fermentation process. In recent years, non-Saccharomyces yeasts such as Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, and Pichia kluyveri have become popular with regard to winemaking and improved wine quality. For that reason, several manufacturers started to offer commercially available strains of these non-Saccharomyces species. P. kluyveri stands out, mainly due to its contribution to wine aroma, glycerol, ethanol yield, and killer factor. The metabolism of the yeast allows it to increase volatile molecules such as esters and varietal thiols (aroma-active compounds), which increase the quality of specific varietal wines or neutral ones. It is considered a low- or non-fermentative yeast, so subsequent inoculation of a more fermentative yeast such as Saccharomyces cerevisiae is indispensable to achieve a proper fermented alcohol. The impact of P. kluyveri is not limited to the grape wine industry; it has also been successfully employed in beer, cider, durian, and tequila fermentation, among others, acting as a promising tool in those fermentation processes. Although no Pichia species other than P. kluyveri is available in the regular market, several recent scientific studies show interesting improvements in some wine quality parameters such as aroma, polysaccharides, acid management, and color stability. This could motivate yeast manufacturers to develop products based on those species in the near future.

scholarly journals Secretion of killer toxin encoded on the linear DNA plasmid pGKL1 from Saccharomyces cerevisiae.

1990 ◽  
Vol 265 (28) ◽  
pp. 17274-17280
Author(s):  
M Tokunaga ◽  
A Kawamura ◽  
K Kitada ◽  
F Hishinuma
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Killer Toxin ◽  
Linear Dna ◽  
Dna Plasmid

scholarly journals Site-Directed Mutagenesis of the Heterotrimeric Killer Toxin Zymocin Identifies Residues Required for Early Steps in Toxin Action

2014 ◽  
Vol 80 (20) ◽  
pp. 6549-6559 ◽  
Author(s):  
Sabrina Wemhoff ◽  
Roland Klassen ◽  
Friedhelm Meinhardt
Keyword(s):  
Kluyveromyces Lactis ◽  
Killer Toxin ◽  
Cysteine Residue ◽  
Site Directed Mutagenesis ◽  
Target Cells ◽  
Directed Mutagenesis ◽  
Content Type ◽  
Protein Toxin ◽  
Assisted Delivery

ABSTRACTZymocin is aKluyveromyces lactisprotein toxin composed of αβγ subunits encoded by the cytoplasmic virus-like element k1 and functions by αβ-assisted delivery of the anticodon nuclease (ACNase) γ into target cells. The toxin binds to cells' chitin and exhibits chitinase activityin vitrothat might be important during γ import.Saccharomyces cerevisiaestrains carrying k1-derived hybrid elements deficient in either αβ (k1ORF2) or γ (k1ORF4) were generated. Loss of either gene abrogates toxicity, and unexpectedly, Orf2 secretion depends on Orf4 cosecretion. Functional zymocin assembly can be restored by nuclear expression of k1ORF2 or k1ORF4, providing an opportunity to conduct site-directed mutagenesis of holozymocin. Complementation required active site residues of α's chitinase domain and the sole cysteine residue of β (Cys250). Since βγ are reportedly disulfide linked, the requirement for the conserved γ C231 was probed. Toxicity of intracellularly expressed γ C231A indicated no major defect in ACNase activity, while complementation of k1ΔORF4 by γ C231A was lost, consistent with a role of β C250 and γ C231 in zymocin assembly. To test the capability of αβ to carry alternative cargos, the heterologous ACNase fromPichia acaciae(P. acaciaeOrf2 [PaOrf2]) was expressed, along with its immunity gene, in k1ΔORF4. While efficient secretion of PaOrf2 was detected, suppression of the k1ΔORF4-derived k1Orf2 secretion defect was not observed. Thus, the dependency of k1Orf2 on k1Orf4 cosecretion needs to be overcome prior to studying αβ's capability to deliver other cargo proteins into target cells.

Killer toxin of Pichia membranifaciens and its possible use as a biocontrol agent against grey mould disease of grapevine

Microbiology ◽  
2004 ◽  
Vol 150 (8) ◽  
pp. 2527-2534 ◽  
Author(s):  
A. Santos ◽  
D. Marquina
Keyword(s):  
Killer Toxin ◽  
Grey Mould ◽  
Toxin Production ◽  
Killer Activity ◽  
Pichia Membranifaciens ◽  
Sds Page ◽  
Toxin Protein ◽  
Killing Action ◽  
Killer Protein ◽  
Potential Use

The use of Pichia membranifaciens CYC 1106 killer toxin against Botrytis cinerea was investigated. This strain exerted a broad-specificity killing action against other yeasts and fungi. At pH 4, optimal killer activity was observed at temperatures up to 20 °C. At 25 °C the toxic effect was reduced to 70 %. The killer activity was higher in acidic medium. Above about pH 4·5 activity decreased sharply and was barely noticeable at pH 6. The killer toxin protein from P. membranifaciens CYC 1106 was purified to electrophoretic homogeneity. SDS-PAGE of the purified killer protein indicated an apparent molecular mass of 18 kDa. Killer toxin production was stimulated in the presence of non-ionic detergents. The toxin concentrations present in the supernatant during optimal production conditions exerted a fungicidal effect on a strain of B. cinerea. The symptoms of infection and grey mould observed in Vitis vinifera plants treated with B. cinerea were prevented in the presence of purified P. membranifaciens killer toxin. The results obtained suggest that P. membranifaciens CYC 1106 killer toxin is of potential use in the biocontrol of B. cinerea.

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