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At least 19 recordsLinked to original sources
PubMed1995

Biocontrol of mold growth in high-moisture wheat stored under airtight conditions by Pichia anomala, Pichia guilliermondii, and Saccharomyces cerevisiae.

Pichia anomala inhibits the growth of Penicillium roqueforti and Aspergillus candidus on agar. In this investigation, antagonistic activity on agar against 17 mold species was determined. The abilities of Pichia anomala, Pichia guilliermondii, and Saccharomyces cerevisiae to inhibit the growth of the mold Penicillium roqueforti in nonsterile high-moisture wheat were compared by adding 10(3) Penicillium roqueforti spores and different amounts of yeast cells per gram of wheat. Inoculated grain was packed in glass tubes, incubated at 25 degrees C with a restricted air supply, and the numbers of yeast and mold CFU were determined on selective media after 7 and 14 days. Pichia anomala reduced growth on agar plates for all of the mold species tested in a dose-dependent manner. Aspergillus fumigatus and Eurotium amstelodami were the most sensitive, while Penicillium italicum and Penicillium digitatum were the most resistant. Pichia anomala had the strongest antagonistic activity in wheat, with 10(5) and 10(6) CFU/g completely inhibiting the growth of Penicillium roqueforti. Inhibition was least pronounced at the optimum temperature (21 degrees C) and water activity (0.95) for the growth of Penicillium roqueforti. Pichia guilliermondii slightly reduced the growth of Penicillium roqueforti in wheat inoculated with 10(5) and 10(6) yeast CFU/g. S. cerevisiae inhibited mold growth only weakly at the highest inoculum level. Pichia anomala grew from 10(3) to 10(7) CFU/g of wheat in 1 week. To reach the same level, Pichia guilliermondii had to be inoculated at 10(4) CFU while S. cerevisiae required an inoculum of 10(5) CFU to reach 10(7) CFU/g of wheat.

Fungi↗
PubMed2025

Taxogenomic analysis of Pichia senei sp. nov. and new insights into hybridization events in the Pichia cactophila species complex.

Three strains of a novel yeast species were isolated from necrotic cactus tissues of Cereus saddianus and Micranthocereus dolichospermaticus and from phytotelmata of Bromelia karatas. DNA sequence analysis of the Internal Transcribed Spacer (ITS) region and D1/D2 domains of the large subunit ribosomal RNA, along with whole genome phylogenomic analysis, showed that this yeast is most closely related to Pichia insulana, Pichia cactophila, and Pichia inconspicua. The new species differs by 10-13 nucleotide substitutions from these species in D1/D2 sequences and exhibits <90% genome-wide average nucleotide identity to them. The name Pichia senei sp. nov. is proposed for the novel species, which is homothallic and produces asci with one to four hat-shaped ascospores. The holotype is CBS 16311 (MycoBank MB 858723). Taxogenomic analyses of the P. cactophila species complex, including P. senei, provide new insights about the hybridizations events that shaped this group. Pichia insulana and P. inconspicua are identified as the parental lineages that originated P. cactophila, and P. senei also appears closely related to one of the progenitors of P. inconspicua. We assess phylogeny, heterozygosity, and ploidy to explore the processes shaping diversity, showing how genomic data support yeast species delimitation and reveal complex hybridization.

Phylogeny↗
PubMed1976

Heterothallism in Pichia kudriavzevii and Pichia terricola.

Pichia kudriavzevii and P. terricola were found to be heterothallic, but not interfertile with one another; nor did they mate with P. membranaefaciens, P. scutulata, Candida lambica, C. diversa, C. ingens, C. silvae, C. valida, C. vini, C. norvegensis, or Torulopsis inconspicua. Limited conjugation occurred between mating types of P. kudriavzevii and C. krusei and conjugation and sporulation occurred in mixtures with C. sorbosa. The data indicate C. krusei and C. sorbosa to be the same species and to represent imperfect forms of P. kudriavzevii.

Ascomycota↗
PubMedDate not supplied

[Conditions and characteristics of the early stages of the sexual interaction of Pichia amethionina var. amethionina and Pichia amethionina var. pachycereana yeast cells].

Pichia amethionina varieties have different sensitivity to the acidity of the medium: P. amethionina var. amethionina has the optimum pH 3 for agglutination and 5 for conjugation; P. amethionina var. pachycereana has the optimum pH 4-6 for agglutination and 6 for conjugation. The optimum temperature for the both organisms is 24 degrees C. Under the optimum conditions, P. amethionina var. amethionina has a 98% agglutination and a 28% conjugation; for P. amethionina var. pachycereana, these values are 83 and 55%, respectively. The agglutination of the two varieties does not depend on the presence of glucose. The conjugation of P. amethionina var. pachycereana does not occur in a medium without glucose while the conjugation of P. amethionina var. amethionina is reduced twofold under these conditions. Inositol, biotin and folic acid produce positive effect on the conjugation of P. amethionina var. pachycereana, but only folic acid favours the conjugation of P. amethionina var. amethionina. No differences have been found between the varieties by staining their cells and zygotes with FITC-ConA.

Agglutination↗
PubMed1976

Serological relationships among some Pichia species.

Antigenic analyses of five species of the genus Pichia were carried out for taxonomic study by the slide agglutination method using monospecific and absorbed antisera and the agglutinin absorption technique. Comparative studies were also performed with a few strains of each of the same species and their classifications are discussed with respect to the antigenic structures and the patterns of proton magnetic resonance (PMR) spectra of their cell wall polysaccharides. ichia delftensis and Pichia zaruensis possessed thermostable antigens 1,2,5 and 11, and the former had also thermoabile antigen m. Both species were closely related to Candida krusei. Pichia toletana possessed thermostable antigens 1,2,5,11,17 and 49. Pichia bovis contained thermostable antigens 1,2,14,15,16,20 and 21, and it was related to most species of the genus Hansenula, although assimilation of potassium nitrate was negative. Finally, Pichia etchellsii possessed thermostable antigens 1,2,3,4,9 and 14, and was closely related to Pichia vini. Patterns of PMR spectra of mannans of these species also supported their serological relationships. Therfore, P. delftensis, P. zaruensis and P. etchellsii are considered to be the synonyms of Pichia fluxuum, Pichia dispora and P. vini respectively, although P. toletanan and P. bovis are independent species.

Antigens, Fungal↗
PubMed1983

Pichia euphorbiae sp. nov., a new haploid heterothallic yeast species.

Representatives of an undescribed, haploid, heterothallic yeast species, Pichia euphorbiae, have been recovered from insect-infested specimens of Euphorbia ingens. The new species is considered to be another representative of the apparently related group of heterothallic species, comprising Pichia rhodanensis, Pichia wickerhamii, Pichia veronae, Pichia amylophila, Pichia mississippiensis and Pichia meyerae. While no mating response was observed in interspecific mixtures of the mating types of the new species with the mating types of the first five mentioned species, a sexual response, manifested by the formation of non-sporulating zygotes, was nevertheless detected with the mating types of P. meyerae. Prototrophic hybrids obtained by crossing auxotrophic mutants of the mating types of P. euphorbiae and P. meyerae were found to be only partial recombinants. A description of the new species is given.

Ascomycota↗
PubMed1997

Assembly of human prolyl 4-hydroxylase and type III collagen in the yeast pichia pastoris: formation of a stable enzyme tetramer requires coexpression with collagen and assembly of a stable collagen requires coexpression with prolyl 4-hydroxylase.

Prolyl 4-hydroxylase, the key enzyme of collagen synthesis, is an alpha2beta2 tetramer, the beta subunit of which is protein disulfide isomerase (PDI). Coexpression of the human alpha subunit and PDI in Pichia produced trace amounts of an active tetramer. A much higher, although still low, assembly level was obtained using a Saccharomyces pre-pro sequence in PDI. Coexpression with human type III procollagen unexpectedly increased the assembly level 10-fold, with no increase in the total amounts of the subunits. The recombinant enzyme was active not only in Pichia extracts but also inside the yeast cell, indicating that Pichia must have a system for transporting all the cosubstrates needed by the enzyme into the lumen of the endoplasmic reticulum. The 4-hydroxyproline-containing procollagen polypeptide chains were of full length and formed molecules with stable triple helices even though Pichia probably has no Hsp47-like protein. The data indicate that collagen synthesis in Pichia, and probably also in other cells, involves a highly unusual control mechanism, in that production of a stable prolyl 4-hydroxylase requires collagen expression while assembly of a stable collagen requires enzyme expression. This Pichia system seems ideal for the high-level production of various recombinant collagens for numerous scientific and medical purposes.

Collagen↗
PubMed2000

Isolation and characterization of the TIM10 homologue from the yeast Pichia sorbitophila: a putative component of the mitochondrial protein import system.

The Saccharomyces cerevisiae TIM10 gene encodes one of the few essential mitochondrial proteins that are required for the import of nuclear-encoded precursor proteins from the cytosol and their subsequent sorting into the different mitochondrial compartments. We have isolated and characterized a putative homologue of TIM10 from the halotolerant yeast Pichia sorbitophila. The Pichia TIM10 gene encodes a protein of 90 amino acids with 66% identity to S. cerevisiae Tim10p. It was capable of suppressing the temperature sensitivity of tim10-1 mutant in S. cerevisiae, suggesting that Pichia TIM10 is both a functional and structural homologue of S. cerevisiae TIM10. The putative Pichia TIM10 gene product contains all the four conserved cysteine residues and the two CX(3)C motifs typical of the Tim family proteins in the mitochondrial intermembrane space. Using anti-Tim10p serum, Western blots detected a protein of about 10 kDa, suggesting that the Pichia Tim10p is a mitochondrial protein. The results suggest that mitochondrial import and sorting systems might be also strongly conserved in other fungi. The coding sequence of the P. sorbitophila TIM10 has been deposited in the EMBL Nucleotide Sequence Database under Accession No. AJ243940.

Amino Acid Sequence↗
PubMed1991

DNA relatedness among saturn-spored yeasts assigned to the genera Williopsis and Pichia.

Saturn-spored species assigned to the genera Williopsis and Pichia were compared from extent of nuclear DNA complementarity. Of the Pichia spp., four were recognized as distinct taxa: P. dispora, P. saitoi, P. zaruensis and Pichia sp. nov. Among Williopsis spp., the following were accepted: W. californica, W. mucosa comb. nov., W. pratensis, W. saturnus var. saturnus, W. saturnus var. mrakii comb. nov., W. saturnus var. sargentensis comb. nov., W. saturnus var. subsufficiens comb. nov. and Williopsis sp. nov. The new Pichia and Williopsis species are described elsewhere. Moderate (36-68%) DNA relatedness was detected between the former Pichia sargentensis and varieties of W. saturnus again demonstrating that nitrate assimilation is not a reliable criterion for separating yeast species.

DNA, Fungal↗
PubMed1984

Synonomy of the yeast genera Hansenula and Pichia demonstrated through comparisons of deoxyribonucleic acid relatedness.

The relationship between the genera Hansenula and Pichia was examined through comparisons of DNA relatedness among phenotypically similar species. Hansenula minuta and Pichia lindneri showed 75% DNA base sequence complementarity. In other comparisons, H. nonfermentans was found to share nearly 50% of its DNA sequences with both H. minuta and P. lindneri. Because of the high degree of relatedness observed, it is proposed that ability to assimilate nitrate, the sole distinction between Hansenula and Pichia, is of insufficient taxonomic value for the reliable separation of either species or genera. Hat-spored species of Hansenula H. et P. Sydow 1919 are being transferred to Pichia Hansen 1904. Species of Hansenula and Pichia with Saturn-shaped ascospores will be transferred to the genus Williopsis.

Ascomycota↗
PubMed1993

Cloning and improving the expression of Pichia stipitis xylose reductase gene in Saccharomyces cerevisiae.

The intact Pichia stipitis xylose reductase gene (XR) has been cloned and expressed in Saccharomyces cerevisiae. The possible further improvement of the expression of the Pichia gene in the new host was studied. To improve the expression of the XR gene in yeast (Saccharomyces cerevisiae), its 5'noncoding sequence containing the genetic elements for transcription and translation was systematically replaced by that from the yeast genes. It was found that the Pichia genetic signal for transcription of XR is more effective than the yeast TRP5 promoter, but is about half as effective as the yeast strong promoter of the alcohol dehydrogenase gene (ADC1). However, the nucleotide sequence immediately adjacent to the initiation codon of XR, which controls the translation of the gene product, seemed to be five times less effective than the corresponding sequence of the ADC1 gene. By totally replacing its 5'-noncoding sequence with that of the yeast ADC1 gene, the expression of XR in yeast was found to be nearly ten times higher. Furthermore, the cloned Pichia XR described in this article contains very little of its 3'-noncoding sequence. In order to study whether the 3'-noncoding sequence is important to its expression in S. cerevisiae, the intact 3'-noncoding sequences of the yeast xylulokinase gene was spliced to the 3' end of the PADC1-XR structural gene. This latter modification has resulted in a twofold further increase in the expression of the Pichia XR in yeast.

Alcohol Dehydrogenase↗
PubMed1991

Genetic transformation of xylose-fermenting yeast Pichia stipitis. Scientific note.

A plasmid-mediated transformation system has been developed for the xylose-fermenting yeast Pichia stipitis. We found that plasmid vectors containing the Saccharomyces cerevisiae 2 mu replicon and the kanamycin resistance gene (KmR) could be introduced into the Pichia cells and maintained as extrachromosomal elements. Pichia transformants containing such vectors will be resistant to the antibiotic geneticin that can be inactivated by the protein product of KmR. Plasmids identical to those used for transformation can be recovered from the Pichia transformants. Protocols for transformation of P. stipitis by the CaCl2-polyethylene glycol-protoplast process or by direct electroporation of intact Pichia cells have both been developed.

Escherichia coli↗
PubMed1998

High-level expression of cockroach allergen, Bla g 4, in Pichia pastoris.

Exposure to cockroach allergens is a risk factor for allergic disease and has been linked to an increase in asthma morbidity among cockroach-sensitive inner-city children. Bla g 4 is a ligand-binding protein (or calycin) that causes IgE antibody responses in 40% to 60% of patients allergic to cockroaches. Recombinant Bla g 4 was expressed in Escherichia coli as an 18 kd protein but provided poor yields (only 0.25 mg/L culture). To improve yields, Bla g 4 was expressed in the Pichia pastoris yeast system as a 23 kd secreted protein at concentrations of 50 mg allergen/L. By cross-inhibition radioimmunoassay, Bla g 4 expressed in E. coli or P. pastoris provided overlapping inhibition curves. Both allergen preparations bound comparable levels of serum IgE antibody and showed similar skin test reactivity in individuals allergic to cockroaches (10[-1] to 10[-3] microg/ml). Deglycosylation of Pichia-expressed Bla g 4 with endoglycosidase F resulted in an 18 to 20 kd doublet, and liquid chromatography-mass spectrometry results suggested that the 20 kd band contained residual sugar residues. Both glycosylated and deglycosylated Pichia Bla g 4 showed comparable inhibition of IgE antibody binding in radioimmunoassay. Pichia-produced Bla g 4 had the same antigenic reactivity as that produced in E. coli, and glycosylation had no effect on IgE antibody binding. The high yield of Bla g 4 obtained in the Pichia system will facilitate studies on the structure and function of calycin allergens and on the immune response of asthma patients to cockroach allergens.

Allergens↗
PubMed1998

Expression of human interleukin-17 in Pichia pastoris: purification and characterization.

A Pichia pastoris expression clone has been developed to produce the human cytokine interleukin-17 (hIL-17). Characterization of purified recombinant hIL-17 made with this clone demonstrated that it shared many characteristics with hIL-17 produced in mammalian cells. The hIL-17 produced in Pichia had the correct N-terminus of natural mature hIL-17 and a glycosylation pattern similar to hIL-17 produced in mammalian cells; both Pichia and human cells add approximately 5 kDa of sugars via N-linked glycosylation and both express a mixture of the glycosylated and nonglycosylated forms. Gel filtration provides evidence that the Pichia produced hIL-17 exists as a dimer in solution. A combination of cation-exchange and gel-filtration chromatography yielded 3.5 mg of highly purified and biologically active hIL-17 from a 10-liter fermentation. These results show that P. pastoris is a useful system to produce recombinant hIL-17 in structure/function studies of this molecule.

Amidohydrolases↗
PubMed1991

Differential toxinogenesis in the genus Pichia detected by an anti-yeast killer toxin monoclonal antibody.

The differential toxinogenesis of 25 isolates belonging to species of the potential yeast killer genus Pichia that were previously classified in the genus Hansenula was comparatively demonstrated by two serologic techniques (indirect immunofluorescence and double immunodiffusion) by using a monoclonal antibody against a yeast killer toxin produced by a selected strain of Pichia anomala (UCSC 25F). The killer phenotypes of the Pichia isolates were evaluated by their ability to kill each other. The results, although of insufficient taxonomic value for a reliable separation of either species or genera, attest to the genomic heterogeneity for the killer character in the genus Pichia as well as the presumptive dual killer/sensitive identity for each single isolate.

Antibodies, Monoclonal↗
PubMed2000

Four new yeasts in the Pichia anomala clade.

Four new yeasts are described that were recognized as novel from nucleotide substitutions in domain D1/D2 of 26S rDNA, a region that is sufficiently divergent to allow resolution of most ascomycetous yeast species. The new species and their type strains are as follows: Pichia maclurae NRRL Y-5377T (= CBS 8671T); Pichia misumaiensis NRRL Y-17389T (= CBS 8062T); Candida mycetangii NRRL Y-6843T (= CBS 8675T); and Candida ulmi NRRL YB-2694T (= CBS 8670T). The two Pichia species form spherical ascospores and are heterothallic. Phylogenetic analysis of domain D1/D2 sequences placed the four new species in the Pichia anomala clade.

Candida↗
PubMed1975

The entry of D-ribose into some yeasts of the genus Pichia.

The utilization of D-ribose by yeasts of the genus Pichia was examined with respect to aerobic growth, respiration and entry of ribose into the cells. Pichia etchellsii (CBS2011) could respire D-ribose, but not use it for aerobic growth. Pichia fermentans (CBS187) neither respired nor grew on D-ribose, though it entered the cells of this yeast either by simple diffusion, or possibly, by the D-glucose carrier, this having a very low affinity for D-ribose. Pichia pinus (CBS5097) respired and grew on D-ribose; kinetic evidence is given for this yeast having two ribose carriers, one inducible and the other constitutive.

Aerobiosis↗