Статті в журналах: "Yeast Cell Surface"

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Bae, Jungu, Kouichi Kuroda, and Mitsuyoshi Ueda. "Proximity Effect among Cellulose-Degrading Enzymes Displayed on the Saccharomyces cerevisiae Cell Surface." Applied and Environmental Microbiology 81, no. 1 (October 10, 2014): 59–66. http://dx.doi.org/10.1128/aem.02864-14.

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ABSTRACTProximity effect is a form of synergistic effect exhibited when cellulases work within a short distance from each other, and this effect can be a key factor in enhancing saccharification efficiency. In this study, we evaluated the proximity effect between 3 cellulose-degrading enzymes displayed on theSaccharomyces cerevisiaecell surface, that is, endoglucanase, cellobiohydrolase, and β-glucosidase. We constructed 2 kinds of arming yeasts through genome integration: ALL-yeast, which simultaneously displayed the 3 cellulases (thus, the different cellulases were near each other), and MIX-yeast, a mixture of 3 kinds of single-cellulase-displaying yeasts (the cellulases were far apart). The cellulases were tagged with a fluorescence protein or polypeptide to visualize and quantify their display. To evaluate the proximity effect, we compared the activities of ALL-yeast and MIX-yeast with respect to degrading phosphoric acid-swollen cellulose after adjusting for the cellulase amounts. ALL-yeast exhibited 1.25-fold or 2.22-fold higher activity than MIX-yeast did at a yeast concentration equal to the yeast cell number in 1 ml of yeast suspension with an optical density (OD) at 600 nm of 10 (OD10) or OD0.1. At OD0.1, the distance between the 3 cellulases was greater than that at OD10 in MIX-yeast, but the distance remained the same in ALL-yeast; thus, the difference between the cellulose-degrading activities of ALL-yeast and MIX-yeast increased (to 2.22-fold) at OD0.1, which strongly supports the proximity effect between the displayed cellulases. A proximity effect was also observed for crystalline cellulose (Avicel). We expect the proximity effect to further increase when enzyme display efficiency is enhanced, which would further increase cellulose-degrading activity. This arming yeast technology can also be applied to examine proximity effects in other diverse fields.

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Nayyar, Ashima, Graeme Walker, Elisabetta Canetta, Forbes Wardrop, and Ashok K. Adya. "Influence of Cell Surface and Nanomechanical Properties on the Flocculation Ability of Industrial Saccharomyces cerevisiae Strains." Journal of Food Research 6, no. 5 (August 2, 2017): 1. http://dx.doi.org/10.5539/jfr.v6n5p1.

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In the past few years, atomic force microscopy (AFM) has provided novel information on the ultrastructural and nanomechanical properties of yeast cell walls that play a major role in determining the flocculation characteristics of the yeasts. In this study, we used AFM to visualize at the nanoscale the cell surface topography and to determine cell wall nanomechanical properties (e.g. elasticity and adhesion) of different strains of S. cerevisiae employed for brewing, winemaking and fuel alcohol production. Cell surface topography was found to correlate with the flocculation behaviour of these strains during their late stationary phase, with the cell surface of flocculent cells being rougher than that of weakly flocculent cells. The elastic modulus of the yeast cell walls showed that weakly flocculent strains had a more rigid cell wall than highly flocculent strains. This difference in elasticity seemed to have an effect on the adhesive properties of the yeast cell walls, with weakly flocculent yeasts displaying lower adhesion energy than the highly flocculent strains. These findings seem to indicate that yeast cell surface nanomechanical properties play an important role in governing flocculation.

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Shibasaki, Seiji, and Mitsuyoshi Ueda. "Progress of Molecular Display Technology Using Saccharomyces cerevisiae to Achieve Sustainable Development Goals." Microorganisms 11, no. 1 (January 3, 2023): 125. http://dx.doi.org/10.3390/microorganisms11010125.

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In the long history of microorganism use, yeasts have been developed as hosts for producing biologically active compounds or for conventional fermentation. Since the introduction of genetic engineering, recombinant proteins have been designed and produced using yeast or bacterial cells. Yeasts have the unique property of expressing genes derived from both prokaryotes and eukaryotes. Saccharomyces cerevisiae is one of the well-studied yeasts in genetic engineering. Recently, molecular display technology, which involves a protein-producing system on the yeast cell surface, has been established. Using this technology, designed proteins can be displayed on the cell surface, and novel abilities are endowed to the host yeast strain. This review summarizes various molecular yeast display technologies and their principles and applications. Moreover, S. cerevisiae laboratory strains generated using molecular display technology for sustainable development are described. Each application of a molecular displayed yeast cell is also associated with the corresponding Sustainable Development Goals of the United Nations.

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4

INOKUMA, Kentaro, and Tomohisa HASUNUMA. "Evolution of Yeast Cell Surface Engineering." Oleoscience 22, no. 3 (2022): 99–105. http://dx.doi.org/10.5650/oleoscience.22.99.

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Bagnat, M., and K. Simons. "Cell surface polarization during yeast mating." Proceedings of the National Academy of Sciences 99, no. 22 (October 8, 2002): 14183–88. http://dx.doi.org/10.1073/pnas.172517799.

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Shimoi, Hitoshi, Kazutoshi Sakamoto, Masaki Okuda, Ratchanee Atthi, Kazuhiro Iwashita, and Kiyoshi Ito. "The AWA1 Gene Is Required for the Foam-Forming Phenotype and Cell Surface Hydrophobicity of Sake Yeast." Applied and Environmental Microbiology 68, no. 4 (April 2002): 2018–25. http://dx.doi.org/10.1128/aem.68.4.2018-2025.2002.

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ABSTRACT Sake, a traditional alcoholic beverage in Japan, is brewed with sake yeasts, which are classified as Saccharomyces cerevisiae. Almost all sake yeasts form a thick foam layer on sake mash during the fermentation process because of their cell surface hydrophobicity, which increases the cells' affinity for bubbles. To reduce the amount of foam, nonfoaming mutants were bred from foaming sake yeasts. Nonfoaming mutants have hydrophilic cell surfaces and no affinity for bubbles. We have cloned a gene from a foam-forming sake yeast that confers foaming ability to a nonfoaming mutant. This gene was named AWA1 and structures of the gene and its product were analyzed. The N- and C-terminal regions of Awa1p have the characteristic sequences of a glycosylphosphatidylinositol anchor protein. The entire protein is rich in serine and threonine residues and has a lot of repetitive sequences. These results suggest that Awa1p is localized in the cell wall. This was confirmed by immunofluorescence microscopy and Western blotting analysis using hemagglutinin-tagged Awa1p. Moreover, an awa1 disruptant of sake yeast was hydrophilic and showed a nonfoaming phenotype in sake mash. We conclude that Awa1p is a cell wall protein and is required for the foam-forming phenotype and the cell surface hydrophobicity of sake yeast.

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7

Thiebault, F., and J. Coulon. "Influence of carbon source and surface hydrophobicity on the aggregation of the yeastKluyveromyces bulgaricus." Canadian Journal of Microbiology 51, no. 1 (January 1, 2005): 91–94. http://dx.doi.org/10.1139/w04-106.

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Aggregation of the yeast Kluyveromyces bulgaricus is mediated by the galactose-specific lectin KbCWL1. This lectin contains hydrophobic amino acids and its activity is calcium dependent. A specific fluorescent probe, 1-anilinonaphthalene-8-sulfonic acid in the free acid form (ANS; Sigma Chemical Co., St. Louis, Missouri), was used to study the hydrophobic areas on the cellular surface of K. bulgaricus. Changes in surface hydrophobicity during the growth and aggregation of yeast cells were studied. Surface hydrophobicity increased during growth and depended on the amount of yeast cells in the culture medium. During growth, the size of the hydrophobic areas on the cell surface was measured using ANS and was found to increase with the percentage of flocculating yeasts. Our results strongly suggest that the hydrophobic areas of the cell walls of yeast cells are involved in the aggregation of K. bulgaricus.Key words: aggregation, carbon source, fluorescence probe, hydrophobicity, yeast.

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8

Shipingana, N. N., N. Raghu, S. Veerana Gowda, T. S. Gopenath, M. S. Ranjith, A. Gnanasekaran, M. Karthikeyan, et al. "Cell signaling in yeast: A mini review." Journal of Biomedical Sciences 5, no. 2 (April 17, 2019): 18–22. http://dx.doi.org/10.3126/jbs.v5i2.23634.

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Background: Understanding cellular mechanism of communication is the main goal of systems biology. Unicellular yeasts are effective model to understand the molecular interactions that generate cell polarity induced by external inputs. The mechanisms of many extracellular stimuli are induced by complexes of cell surface receptors, G proteins. The mechanisms of many extracellular stimuli are induced by complexes of cell surface receptors, G proteins and mitogen activated protein (MAP) kinase complexes. Many components, their interrelationships, and their regulators of these mechanisms were initially identified in yeast. A complex web of sensing mechanisms and cooperation among signaling networks such as a cyclic adenosine monophosphate dependent protein kinase, mitogen-activated protein kinase cascade and 5-adenosine monophosphate activated protein kinase induce various changes in physiology, cell polarity, cell cycle progression and gene expression to achieve differentiation. Ras-cAMP pathway explained in yeast model with signalling function of the oncogenic mammalian Ras protein. So studies on yeast cells may enlighten some underlying mechanism which will be beneficial to understand the mechanisms of disease.

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9

Shibasaki, Seiji, Yuki Nakatani, Kazuaki Taketani, Miki Karasaki, Kiyoshi Matsui, Mitsuyoshi Ueda, and Tsuyoshi Iwasaki. "Construction of HGF-Displaying Yeast by Cell Surface Engineering." Microorganisms 10, no. 7 (July 7, 2022): 1373. http://dx.doi.org/10.3390/microorganisms10071373.

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Hepatocyte growth factor (HGF) has been investigated as a regulator for immune reactions caused by transplantation and autoimmune diseases and other biological functions. Previous studies demonstrated that cDNA-encoding HGF administration could inhibit acute graft-versus-host disease (GVHD) after treatment via hematopoietic stem cell transplantation. This study aimed to show the preparation of HGF protein on yeast cell surfaces to develop a tool for the oral administration of HGF to a GVHD mouse model. In this study, full-length HGF and the heavy chain of HGF were genetically fused with α-agglutinin and were successfully displayed on the yeast cell surface. This study suggested that yeast cell surface display engineering could provide a novel administration route for HGF.

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10

Coleman, David A., Soon-Hwan Oh, Xiaomin Zhao, and Lois L. Hoyer. "Heterogeneous distribution of Candida albicans cell-surface antigens demonstrated with an Als1-specific monoclonal antibody." Microbiology 156, no. 12 (December 1, 2010): 3645–59. http://dx.doi.org/10.1099/mic.0.043851-0.

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Despite an abundance of data describing expression of genes in the Candida albicans ALS (agglutinin-like sequence) gene family, little is known about the production of Als proteins on individual cells, their spatial localization or stability. Als proteins are most commonly discussed with respect to function in adhesion of C. albicans to host and abiotic surfaces. Development of a mAb specific for Als1, one of the eight large glycoproteins encoded by the ALS family, provided the opportunity to detect Als1 during growth of yeast and hyphae, both in vitro and in vivo, and to demonstrate the utility of the mAb in blocking C. albicans adhesion to host cells. Although most C. albicans yeast cells in a saturated culture are Als1-negative by indirect immunofluorescence, Als1 is detected on the surface of nearly all cells shortly after transfer into fresh growth medium. Als1 covers the yeast cell surface, with the exception of bud scars. Daughters of the inoculum cells, and sometimes granddaughters, also have detectable Als1, but Als1 is not detectable on cells from subsequent generations. On germ tubes and hyphae, most Als1 is localized proximal to the mother yeast. Once deposited on yeasts or hyphae, Als1 persists long after the culture has reached saturation. Growth stage-dependent production of Als1, coupled with its persistence on the cell surface, results in a heterogeneous population of cells within a C. albicans culture. Anti-Als1 immunolabelling patterns vary depending on the source of the C. albicans cells, with obvious differences between cells recovered from culture and those from a murine model of disseminated candidiasis. Results from this work highlight the temporal parallels for ALS1 expression and Als1 production in yeasts and germ tubes, the specialized spatial localization and persistence of Als1 on the C. albicans cell surface, and the differences in Als1 localization that occur in vitro and in vivo.

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11

UEDA, MITSUYOSHI. "Cell surface layer engineering of the yeast." Kagaku To Seibutsu 35, no. 7 (1997): 525–32. http://dx.doi.org/10.1271/kagakutoseibutsu1962.35.525.

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12

Ueda, Mitsuyoshi, and Atsuo Tanaka. "Cell surface engineering of yeast: Construction of arming yeast with biocatalyst." Journal of Bioscience and Bioengineering 90, no. 2 (January 2000): 125–36. http://dx.doi.org/10.1016/s1389-1723(00)80099-7.

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13

UEDA, MITSUYOSHI, and ATSUO TANAKA. "Cell Surface Engineering of Yeast. Construction of Arming Yeast with Biocatalyst." Journal of Bioscience and Bioengineering 90, no. 2 (2000): 125–36. http://dx.doi.org/10.1263/jbb.90.125.

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14

Braun, Phyllis C. "Nutrient uptake byCandida albicans: the influence of cell surface mannoproteins." Canadian Journal of Microbiology 45, no. 5 (July 1, 1999): 353–59. http://dx.doi.org/10.1139/w99-035.

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Numerous ultrastructural and biochemical analyses have been performed to characterize the cell wall composition and structure of Candida albicans. However, little investigation has focused on how subtle differences in cell wall structure influence the intracellular transport of amino acids and monosaccharides. In this study C. albicans 4918 and ATCC 10231 were grown in culture conditions capable of modifying surface mannoproteins and induced surface hydrophobic or hydrophilic yeast cell wall states. Subcultures of these hydrophobic and hydrophilic yeasts were subsequently incubated with one of seven L-[3H] amino acids: glycine, leucine, proline, serine, aspartic acid, lysine, or arginine. The transport of [3H] mannose and [3H] N-acetyl-D-glucosamine were also investigated. This study revealed significant strain differences (P [Formula: see text] 0.05) between hydrophilic and hydrophobic yeast transport of these nutrients throughout a 2 h incubation. Hydrophilic cultures of 4918 and ATCC 10231 transported nearly two times more (pmol mg-1dry weight) proline, mannose, and N-acetyl-D-glucosamine than hydrophobic yeast. Hydrophobic cultures preferentially incorporated serine and aspartic acid in both these strains. Strain variation was indicated with the transport of leucine, lysine, and arginine, as follows: experiments showed that hydrophilic 4918 cultures selectively transported leucine, lysine, and arginine, whereas, the hydrophobic ATCC 10231 cultures incorporated these amino acids.Key words: Candida albicans, mannoproteins, amino acid transport.

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15

Sayin, Ismail, Mehmet Kahraman, Fikrettin Sahin, Dilsad Yurdakul, and Mustafa Culha. "Characterization of Yeast Species Using Surface-Enhanced Raman Scattering." Applied Spectroscopy 63, no. 11 (November 2009): 1276–82. http://dx.doi.org/10.1366/000370209789806849.

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Surface-enhanced Raman scattering (SERS) is used for the characterization of six yeast species and six isolates. The sample for SERS analysis is prepared by mixing the yeast cells with a four times concentrated silver colloidal suspension. The scanning electron microscopy (SEM) images show that the strength of the interaction between silver nanoparticles and the yeast cells depends on the biochemical structure of the cell wall. The SERS spectra are used to identify the biochemical structures on the yeast cell wall. It is found that the density of –SH and –NH2 groups might be higher on certain yeast cell walls. Finally, the obtained SERS spectra from yeast is used for the classification of the yeast.

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Gregoire, S., J. Xiao, B. B. Silva, I. Gonzalez, P. S. Agidi, M. I. Klein, K. S. Ambatipudi, et al. "Role of Glucosyltransferase B in Interactions of Candida albicans with Streptococcus mutans and with an Experimental Pellicle on Hydroxyapatite Surfaces." Applied and Environmental Microbiology 77, no. 18 (July 29, 2011): 6357–67. http://dx.doi.org/10.1128/aem.05203-11.

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ABSTRACTCandida albicansand mutans streptococci are frequently detected in dental plaque biofilms from toddlers afflicted with early childhood caries. Glucosyltransferases (Gtfs) secreted byStreptococcus mutansbind to saliva-coated apatite (sHA) and to bacterial surfaces, synthesizing exopolymersin situ, which promote cell clustering and adherence to tooth enamel. We investigated the potential role Gtfs may play in mediating the interactions betweenC. albicansSC5314 andS. mutansUA159, both with each other and with the sHA surface. GtfB adhered effectively to theC. albicansyeast cell surface in an enzymatically active form, as determined by scintillation spectroscopy and fluorescence imaging. The glucans formed on the yeast cell surface were more susceptible to dextranase than those synthesized in solution or on sHA and bacterial cell surfaces (P< 0.05), indicating an elevated α-1,6-linked glucose content. Fluorescence imaging revealed that larger numbers ofS. mutanscells bound toC. albicanscells with glucans present on their surface than to yeast cells without surface glucans (uncoated). The glucans formedin situalso enhancedC. albicansinteractions with sHA, as determined by a novel single-cell micromechanical method. Furthermore, the presence of glucan-coated yeast cells significantly increased the accumulation ofS. mutanson the sHA surface (versusS. mutansincubated alone or mixed with uncoatedC. albicans;P< 0.05). These data reveal a novel cross-kingdom interaction that is mediated by bacterial GtfB, which readily attaches to the yeast cell surface. Surface-bound GtfB promotes the formation of a glucan-rich matrixin situand may enhance the accumulation ofS. mutanson the tooth enamel surface, thereby modulating the development of virulent biofilms.

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Klis, Frans M., Marian de Jong, Stanley Brul, and Piet W. J. de Groot. "Extraction of cell surface-associated proteins from living yeast cells." Yeast 24, no. 4 (2007): 253–58. http://dx.doi.org/10.1002/yea.1476.

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MacDonald, Chris, and Robert C. Piper. "Cell surface recycling in yeast: mechanisms and machineries." Biochemical Society Transactions 44, no. 2 (April 11, 2016): 474–78. http://dx.doi.org/10.1042/bst20150263.

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Sorting internalized proteins and lipids back to the cell surface controls the supply of molecules throughout the cell and regulates integral membrane protein activity at the surface. One central process in mammalian cells is the transit of cargo from endosomes back to the plasma membrane (PM) directly, along a route that bypasses retrograde movement to the Golgi. Despite recognition of this pathway for decades we are only beginning to understand the machinery controlling this overall process. The budding yeast Saccharomyces cerevisiae, a stalwart genetic system, has been routinely used to identify fundamental proteins and their modes of action in conserved trafficking pathways. However, the study of cell surface recycling from endosomes in yeast is hampered by difficulties that obscure visualization of the pathway. Here we briefly discuss how recycling is likely a more prevalent process in yeast than is widely appreciated and how tools might be built to better study the pathway.

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19

Colling, Lisa, Michael Essmann, Cara Hollmer, and Bryan Larsen. "Surface Modifying Substances that Reduce Apparent Yeast Cell Hydrophobicity." Infectious Diseases in Obstetrics and Gynecology 13, no. 3 (2005): 171–77. http://dx.doi.org/10.1080/10647440500068149.

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Conclusions.Several commercially available compounds were able to block binding of styrene microspheres to yeast. Some of the binding activity appeared to be attributable to mannose-containing surface components. These findings have implications for formulating therapeutic products that might block yeast binding to tissues.

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20

Colling, Lisa, Richard N. Carter, Michael Essmann, and Bryan Larsen. "Evaluation of Relative Yeast Cell Surface Hydrophobicity Measured by Flow Cytometry." Infectious Diseases in Obstetrics and Gynecology 13, no. 1 (2005): 43–48. http://dx.doi.org/10.1155/2005/739101.

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Objective:To develop an efficient method for evaluating cell surface hydrophobicity and to apply the method to demonstrate the effects of fungal growth conditions on cell surface properties.Methods:Yeast isolates were suspended in phosphate-buffered saline and mixed with deep blue-dyed polystyrene microspheres. Flow cytometry was used to detect the degree of microsphere binding to yeast cells. Different strains of yeast were compared for intrinsic microsphere binding activity and changes in growth conditions were invoked to modify the relative surface hydrophobicity.Results:Commercially available blue-dyed polystyrene microspheres showed strong fluorescence in the FL3 channel, whereas yeast cells did not show appreciable FL3 fluorescence. Microspheres and yeast were generally distinguishable on the basis of size revealed by forward light scatter. This method showed a wide variation in intrinsic cell surface hydrophobicity amongCandida albicansstrains. Likewise, variation in hydrophobicity of non-albicans yeast species was observed. Growth on solid media, incubation at 25°C, or 250 mg/dl glucose concentration increased hydrophobicity compared with growth in liquid media, incubation at 37°C, or 50 mg/dl glucose, respectively. Growth in1×10−9M estradiol had no appreciable effect on hydrophobicity.Conclusions:Stained latex microspheres fluoresced in the FL3 channel of the flow cytometer and bound to yeast cells to an extent related to the surface hydrophobicity of the yeast. Binding detected by flow cytometry showed that clinical yeast isolates varied in intrinsic binding capacity and this binding ability was altered by different growth conditions. The implications for virulence regulation among yeast isolates are discussed.

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21

Tran, Clara T. H., Alexey Kondyurin, Stacey L. Hirsh, David R. McKenzie, and Marcela M. M. Bilek. "Ion-implanted polytetrafluoroethylene enhances Saccharomyces cerevisiae biofilm formation for improved immobilization." Journal of The Royal Society Interface 9, no. 76 (June 13, 2012): 2923–35. http://dx.doi.org/10.1098/rsif.2012.0347.

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The surface of polytetrafluoroethylene (PTFE) was modified using plasma immersion ion implantation (PIII) with the aim of improving its ability to immobilize yeast. The density of immobilized cells on PIII-treated and -untreated PTFE was compared as a function of incubation time over 24 h. Rehydrated yeast cells attached to the PIII-treated PTFE surface more rapidly, with higher density, and greater attachment strength than on the untreated surface. The immobilized yeast cells were removed mechanically or chemically with sodium hydroxide and the residues left on the surfaces were analysed with Fourier transform infrared spectroscopy-attenuated total reflection (FTIR-ATR) and X-ray photoelectron spectroscopy (XPS). The results revealed that the mechanism of cell attachment on both surfaces differs and a model is presented for each. Rapid attachment on the PIII-treated surface occurs through covalent bonds of cell wall proteins and the radicals on the treated surface. In contrast, on the untreated surface, only physisorbed molecules were found in the residue and lipids were more highly concentrated than proteins. The presence of lipids in the residue was found to be a consequence of damage to the plasma membrane during the rehydration process and the increased cell stress was also apparent by the amount of Hsp12 in the protein residue. The immobilized yeast cells on PIII-treated PTFE were found to be as active as yeast cells in suspension.

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Nakari-Setälä, Tiina, Joana Azeredo, Mariana Henriques, Rosário Oliveira, José Teixeira, Markus Linder, and Merja Penttilä. "Expression of a Fungal Hydrophobin in the Saccharomyces cerevisiae Cell Wall: Effect on Cell Surface Properties and Immobilization." Applied and Environmental Microbiology 68, no. 7 (July 2002): 3385–91. http://dx.doi.org/10.1128/aem.68.7.3385-3391.2002.

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ABSTRACT The aim of this work was to modify the cell surface properties of Saccharomyces cerevisiae by expression of the HFBI hydrophobin of the filamentous fungus Trichoderma reesei on the yeast cell surface. The second aim was to study the immobilization capacity of the modified cells. Fusion to the Flo1p flocculin was used to target the HFBI moiety to the cell wall. Determination of cell surface characteristics with contact angle and zeta potential measurements indicated that HFBI-producing cells are more apolar and slightly less negatively charged than the parent cells. Adsorption of the yeast cells to different commercial supports was studied. A twofold increase in the binding affinity of the hydrophobin-producing yeast to hydrophobic silicone-based materials was observed, while no improvement in the interaction with hydrophilic carriers could be seen compared to that of the parent cells. Hydrophobic interactions between the yeast cells and the support are suggested to play a major role in attachment. Also, a slight increase in the initial adsorption rate of the hydrophobin yeast was observed. Furthermore, due to the engineered cell surface, hydrophobin-producing yeast cells were efficiently separated in an aqueous two-phase system by using a nonionic polyoxyethylene detergent, C12-18EO5.

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Li, Yumei, Lili Lu, Hongmei Wang, Xiaodong Xu та Min Xiao. "Cell Surface Engineering of a β-Galactosidase for Galactooligosaccharide Synthesis". Applied and Environmental Microbiology 75, № 18 (17 липня 2009): 5938–42. http://dx.doi.org/10.1128/aem.00326-09.

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ABSTRACT A novel gene encoding transglycosylating β-galactosidase (BGase) was cloned from Penicillium expansum F3. The sequence contained a 3,036-bp open reading frame encoding a 1,011-amino-acid protein. This gene was subsequently expressed on the cell surface of Saccharomyces cerevisiae EBY-100 by galactose induction. The BGase-anchored yeast could directly utilize lactose to produce galactooligosaccharide (GOS), as well as the by-products glucose and a small quantity of galactose. The glucose was consumed by the yeast, and the galactose was used for BGase expression, thus greatly facilitating GOS synthesis. The GOS yield reached 43.64% when the recombinant yeast was cultivated in yeast nitrogen base-Casamino Acids medium containing 100 g/liter initial lactose at 25°C for 5 days. The yeast cells were harvested and recycled for the next batch of GOS synthesis. During sequential operations, both oligosaccharide synthesis and BGase expression were maintained at high levels with GOS yields of over 40%, and approximately 8 U/ml of BGase was detected in each batch.

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Konnova, S. A., Y. M. Lvov, and R. F. Fakhrullin. "Magnetic halloysite nanotubes for yeast cell surface engineering." Clay Minerals 51, no. 3 (June 2016): 429–33. http://dx.doi.org/10.1180/claymin.2016.051.3.07.

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AbstractHalloysite clay nanotubes are safe and biocompatible nanomaterials and their application in biomaterials is very promising. The microencapsulation of yeast cells in the shell of clay nanotubes modifying their properties was demonstrated here. Each cell was coated with a 200–300 nm-thick tube shell and this coating was not harmful for these cells’ reproduction. Synthesis of magnetic nanoparticles on the surfaces of the nanotubes allowed for magnetic-field manipulation of the coated cells, including their separation. Providing nano-designed shells for biological cells is a step forward in development of ‘cyborg’ microorganisms combining their intrinsic properties with functions added through nano-engineering.

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FUKUDA, Takeshi, Danya ISOGAWA, Madoka TAKAGI, Michiko KATO-MURAI, Hisashi KIMOTO, Hideo KUSAOKE, Mitsuyoshi UEDA, and Shin-ichiro SUYE. "Yeast Cell-Surface Expression of Chitosanase fromPaenibacillus fukuinensis." Bioscience, Biotechnology, and Biochemistry 71, no. 11 (November 23, 2007): 2845–47. http://dx.doi.org/10.1271/bbb.70315.

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Kondo, A., and M. Ueda. "Yeast cell-surface display?applications of molecular display." Applied Microbiology and Biotechnology 64, no. 1 (March 1, 2004): 28–40. http://dx.doi.org/10.1007/s00253-003-1492-3.

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27

Roemer, Terry, and Howard Bussey. "Yeast Kre1p is a cell surface O-glycoprotein." Molecular and General Genetics MGG 249, no. 2 (March 1995): 209–16. http://dx.doi.org/10.1007/bf00290368.

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Hossain, SK Amir, SM Rifat Rahman, Toufiq Ahmed, and Chanchal Mandal. "An overview of yeast cell wall proteins and their contribution in yeast display system." Asian Journal of Medical and Biological Research 5, no. 4 (February 3, 2020): 246–57. http://dx.doi.org/10.3329/ajmbr.v5i4.45261.

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Yeast surface display has become an increasingly popular tool for protein engineering and library screening applications. Although, recent advances have greatly expanded the capability of yeast surface display, the protein display system is still far away from industrial application. One of the major components of a stable, efficient and successful yeast surface display system is cell wall anchor protein with which our desired foreign protein will be attached. We studied 80 different yeast cell wall anchored proteins originated mostly from Saccharomyces cerevisiae and Candida albicans. We studied in details all the cell wall proteins in order to find out suitable cell wall proteins to recommend for the researchers to use in the construction of yeast display system. We considered selective physical properties of different yeast cell wall proteins that are crucial for selecting best suited cell surface anchor proteins which are molecular weight, binding domain of anchor protein, length of amino acid and fusion site. Finally, our studies showed that Ccw11, Ccw12. Cwp1, Cwp2, Dan1, Gas1, Gas5, Exg1, Ycr89, Ecm33, Pga4, Sap9, Sap10, Pst1, Pir1, Pir2, Pir3, Pir4, Cis1, Scw4, Scw6, Bgl2, Uth1, Scw1 are the promising and suitable cell wall anchor proteins could be used in construction of yeast cell surface display system. Additionally, this review presents detailed information about all the cell wall proteins in a single work. The future researchers in this field will be able to construct more efficient yeast display system for recombinant protein production at industrial scale using the knowledge presented in this work. Asian J. Med. Biol. Res. June 2019, 5(4): 246-257

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Lu, Dongdong, Songsong Tang, Yangyang Li, Zhaoqing Cong, Xueji Zhang, and Song Wu. "Magnetic-Propelled Janus Yeast Cell Robots Functionalized with Metal-Organic Frameworks for Mycotoxin Decontamination." Micromachines 12, no. 7 (July 5, 2021): 797. http://dx.doi.org/10.3390/mi12070797.

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Cell robots that transform natural cells into active platforms hold great potential to enrich the biomedical prospects of artificial microrobots. Here, we present Janus yeast cell microrobots (JYC-robots) prepared by asymmetrically coating Fe3O4 nanoparticles (NPs) and subsequent in situ growth of zeolitic imidazolate framework-67 (ZIF-67) on the surface of yeast cells. The magnetic actuation relies on the Fe3O4 NPs wrapping. As the compositions of cell robots, the cell wall with abundant polysaccharide coupling with porous and oxidative ZIF-67 can concurrently remove mycotoxin (e.g., zearalenone (ZEN)). The magnetic propulsion accelerates the decontamination efficiency of JYC-robots against ZEN. Although yeast cells with fully coating of Fe3O4 NPs and ZIF-67 (FC-yeasts) show faster movement than JYC-robots, higher toxin-removal efficacy is observed for JYC-robots compared with that of FC-yeasts, reflecting the vital factor of the yeast cell wall in removing mycotoxin. Such design with Janus modification of magnetic NPs (MNPs) and entire coating of ZIF-67 generates active cell robot platform capable of fuel-free propulsion and enhanced detoxification, offering a new formation to develop cell-based robotics system for environmental remediation.

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BEUCHAT, L. R., B. V. NAIL, R. E. BRACKETT, and T. L. FOX. "Evaluation of a Culture Film (Petrifilm™ YM) Method for Enumerating Yeasts and Molds in Selected Dairy and High-Acid Foods." Journal of Food Protection 53, no. 10 (October 1, 1990): 869–74. http://dx.doi.org/10.4315/0362-028x-53.10.869.

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The Petrifilm™ Yeast and Mold (YM) plate was compared to acidified potato dextrose agar (APDA) and chloramphenicol-supplemented plate count agar (CPCA) using pour- and surface-plating techniques for its ability to recover yeasts and molds from hard and soft cheeses, cottage cheese, yogurt, sour cream, fruit juice, salad dressing, relishes, and tomato-based sauces. Correlation coefficients of Petrifilm™ YM plates versus pour-APDA, surface-APDA, pour-CPCA, and surface-CPCA for recovering total yeasts and molds from a composite of the eight test foods were, respectively, 0.993, 0.993, 0.994, and 0.995. Slope and intercept values for populations detected using Petrifilm™ YM plates versus traditional systems ranged, respectively, from 0.984 to 1.008 and −0.051 to 0.149. The coefficient of variation for total yeast and mold populations recovered on Petrifilm™ YM plates was 1.0% compared to 1.2 to 1.7% for traditional enumeration systems. Regardless of the enumeration system employed or the type of fungal cell, i.e., yeast or mold, being enumerated, significantly (P ≤ 0.05) higher populations were generally detected after 5 d compared to 3 d of incubation. After 5 d of incubation, in no case were yeast or total yeast and mold populations detected in the eight food products using Petrifilm™ YM plates significantly lower than respective populations detected using traditional pour- and surface-plating techniques and media. When Petrifilm™ YM plates were used, significantly higher total yeast and mold populations were detected in 3, 1, and 1 out of eight food products compared to using, respectively, pour-APDA, surface-APDA, and surface-CPCA enumeration systems. The Petrifilm™ YM plate offers an acceptable alternative to traditional methods for enumerating yeasts and molds in the dairy and high-acid products evaluated in this study.

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UEDA, MITSUYOSHI, TOSHIYUKI MURAI, YUMI SHIBASAKI, NAOMI KAMASAWA, MASAKO OSUMI, and ATSUO TANAKA. "Molecular Breeding of Polysaccharide-Utilizing Yeast Cells by Cell Surface Engineering." Annals of the New York Academy of Sciences 864, no. 1 ENZYME ENGINE (December 1998): 528–37. http://dx.doi.org/10.1111/j.1749-6632.1998.tb10374.x.

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32

Watanabe, Yukio, Wataru Aoki, and Mitsuyoshi Ueda. "Improved ammonia production from soybean residues by cell surface-displayed l-amino acid oxidase on yeast." Bioscience, Biotechnology, and Biochemistry 85, no. 4 (December 21, 2020): 972–80. http://dx.doi.org/10.1093/bbb/zbaa112.

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ABSTRACT Ammonia is critical for agricultural and chemical industries. The extracellular production of ammonia by yeast (Saccharomyces cerevisiae) using cell surface engineering can be efficient approach because yeast can avoid growth deficiencies caused by knockout of genes for ammonia assimilation. In this study, we produced ammonia outside the yeast cells by displaying an l-amino acid oxidase with a wide substrate specificity derived from Hebeloma cylindrosporum (HcLAAO) on yeast cell surfaces. The HcLAAO-displaying yeast successfully produced 12.6 m m ammonia from a mixture of 20 proteinogenic amino acids (the theoretical conversion efficiency was 63%). We also succeeded in producing ammonia from a food processing waste, soybean residues (okara) derived from tofu production. The conversion efficiency was 88.1%, a higher yield than reported in previous studies. Our study demonstrates that ammonia production outside of yeast cells is a promising strategy to utilize food processing wastes.

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Wahyuni, I., U. Purwandari, A. Subagio, and N. Nurhayati. "Isolation and identification of gastric acid-tolerant yeast from tapai." Food Research 7, Supplementary 1 (August 15, 2023): 276–82. http://dx.doi.org/10.26656/fr.2017.7(s1).13.

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Lactobacilli and Bifidobacteria are the most extensively employed bacterial strains in marketable probiotic supplements. However, another probiotic was recently developed from yeast screening based on tolerance against gastric acid. This research aimed to isolate yeasts from traditional Indonesian fermented food (tapai). Screening of probiotic yeasts was based on their survival in gastric acid of pH 2.0. Yeast strains were cultured in malt extract agar, and their phenotype and genotype characteristics were identified. Phenotype characteristics were based on yeast cells’ colony, microscopy, and physiology. Meanwhile, genotype characteristics were determined using the PCR-fingerprinting technique to identify the sequence homology compared to the GenBank database and the phylogenetic tree construction. The result showed that SUL and SM isolates have the highest survival on artificial gastric acid of pH 2.0. The SUL isolate from tapai brand “Sumber Madu” has a morphologically wrinkled colony, no pseudo mycelium, white surface colony, and round cell shape. In contrast, the SM isolate from tapai brand “Sari Madu” has a thin wide colony with no pseudo mycelium, turbid white surface, and oval cell shape. After 2 h incubation on gastric acid, SUL and SM isolates grew up to 6.20±0.35 CFU/mL (survival yeast of 82.71%) and 5.75±0.45 CFU/mL (survival yeast of 79.74%), respectively. The SUL isolate was identified as Kodamaea ohmeri, while the SM isolate was identified as Pichia kudriavzevii.

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Bouyx, Clara, Marion Schiavone, and Jean Marie François. "FLO11, a Developmental Gene Conferring Impressive Adaptive Plasticity to the Yeast Saccharomyces cerevisiae." Pathogens 10, no. 11 (November 19, 2021): 1509. http://dx.doi.org/10.3390/pathogens10111509.

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The yeast Saccharomyces cerevisiae has a remarkable ability to adapt its lifestyle to fluctuating or hostile environmental conditions. This adaptation most often involves morphological changes such as pseudofilaments, biofilm formation, or cell aggregation in the form of flocs. A prerequisite for these phenotypic changes is the ability to self-adhere and to adhere to abiotic surfaces. This ability is conferred by specialized surface proteins called flocculins, which are encoded by the FLO genes family in this yeast species. This mini-review focuses on the flocculin encoded by FLO11, which differs significantly from other flocculins in domain sequence and mode of genetic and epigenetic regulation, giving it an impressive plasticity that enables yeast cells to swiftly adapt to hostile environments or into new ecological niches. Furthermore, the common features of Flo11p with those of adhesins from pathogenic yeasts make FLO11 a good model to study the molecular mechanism underlying cell adhesion and biofilm formation, which are part of the initial step leading to fungal infections.

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35

UEDA, Mitsuyoshi, and Atsuo TANAKA. "Novel Molecular Breeding of Yeast by Cell Surface Engineering." JOURNAL OF THE BREWING SOCIETY OF JAPAN 94, no. 11 (1999): 860–67. http://dx.doi.org/10.6013/jbrewsocjapan1988.94.860.

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36

Kondo, Akihiko, Tsutomu Tanaka, Tomohisa Hasunuma, and Chiaki Ogino. "Applications of Yeast Cell-Surface Display in Bio-Refinery." Recent Patents on Biotechnology 4, no. 3 (November 1, 2010): 226–34. http://dx.doi.org/10.2174/187220810793611509.

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37

Ueda, Mitsuyoshi, and Atsuo Tanaka. "Genetic immobilization of proteins on the yeast cell surface." Biotechnology Advances 18, no. 2 (April 2000): 121–40. http://dx.doi.org/10.1016/s0734-9750(00)00031-8.

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38

Harsay, E., and A. Bretscher. "Parallel secretory pathways to the cell surface in yeast." Journal of Cell Biology 131, no. 2 (October 15, 1995): 297–310. http://dx.doi.org/10.1083/jcb.131.2.297.

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Saccharomyces cerevisiae mutants that have a post-Golgi block in the exocytic pathway accumulate 100-nm vesicles carrying secretory enzymes as well as plasma membrane and cell-wall components. We have separated the vesicle markers into two groups by equilibrium isodensity centrifugation. The major population of vesicles contains Bg12p, an endoglucanase destined to be a cell-wall component, as well as Pma1p, the major plasma membrane ATPase. In addition, Snc1p, a synaptobrevin homologue, copurifies with these vesicles. Another vesicle population contains the periplasmic enzymes invertase and acid phosphatase. Both vesicle populations also contain exoglucanase activity; the major exoglucanase normally secreted from the cell, encoded by EXG1, is carried in the population containing periplasmic enzymes. Electron microscopy shows that both vesicle groups have an average diameter of 100 nm. The late secretory mutants sec1, sec4, and sec6 accumulate both vesicle populations, while neither is detected in wild-type cells, early sec mutants, or a sec13 sec6 double mutant. Moreover, a block in endocytosis does not prevent the accumulation of either vesicle species in an end4 sec6 double mutant, further indicating that both populations are of exocytic origin. The accumulation of two populations of late secretory vesicles indicates the existence of two parallel routes from the Golgi to the plasma membrane.

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39

Kono, Keiko, Hiroki Okada, and Yoshikazu Ohya. "Local and Acute Disruption of the Yeast Cell Surface." Cold Spring Harbor Protocols 2016, no. 8 (August 2016): pdb.prot085266. http://dx.doi.org/10.1101/pdb.prot085266.

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40

Zhang, Wei, Bo Zhang, Yu Long Zhang, Da Han, and Yong Liang Zhou. "Trapping Yeast Cells on PDMS Micropillar Array." Advanced Materials Research 476-478 (February 2012): 2096–99. http://dx.doi.org/10.4028/www.scientific.net/amr.476-478.2096.

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A selectively yeast cell trapping and arraying method was presented, utilizing PDMS micropillar array combining with chemical adhesion. Yeast cells were trapped on the top of micropillar array while swept on the flat area for the pinning of liquid/surface contact line on the micropillars and moving on the flat surface. And the modification of poly-L-lysine on the yeast cells improved the immobilization of cells on the surface of PDMS surface. Both of simulation and experiment results shows that by adjusting the diameter of micropillars, the number of yeast cells on each pillars could be controlled. Single cell array was formed with a 8.3 μm micropillar array, and majority yeast numbers of 3,4,5 was got for the 13.7 μm、18.0 μm and 18.8 μm micropillar array.

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41

Buzzini, Pietro, Benedetta Turchetti, Guglielmina Diolaiuti, Carlo D’Agata, and Alessandro Martini. "Culturable yeasts in meltwaters draining from two glaciers in the Italian Alps." Annals of Glaciology 40 (2005): 119–22. http://dx.doi.org/10.3189/172756405781813591.

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AbstractThe meltwaters draining from two glaciers in the Italian Alps contain metabolically active yeasts isolable by culture-based laboratory procedures. The average number of culturable yeast cells in the meltwaters was 10–20 colony-forming units (CFU) L–1, whereas supraglacial stream waters originating from overlying glacier ice contained <1 CFUL–1. Yeast cell number increased as the suspended-sediment content of the water samples increased. Basidiomycetous yeasts represent >80% of isolated strains (Cryptococcus spp. and Rhodotorula spp. were 33.3% and 17.8% of total strains, respectively). Culturable yeasts were psychrotolerant, predominantly obligate aerobes and able to degrade organic macromolecules (e.g. starch, esters, lipids, proteins). To the authors’ knowledge, this is the first study to report the presence of culturable yeasts in meltwaters originating from glaciers. On the basis of these results, it is reasonable to suppose that the viable yeasts observed in meltwaters derived predominantly from the subglacial zone and that they originated from the subglacial microbial community. Their metabolic abilities could contribute to the microbial activity occurring in subglacial environments.

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Liu, Yun, Rui Zhang, Zhongshuai Lian, Shihui Wang, and Aaron T. Wright. "Yeast cell surface display for lipase whole cell catalyst and its applications." Journal of Molecular Catalysis B: Enzymatic 106 (August 2014): 17–25. http://dx.doi.org/10.1016/j.molcatb.2014.04.011.

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43

Shenoy, Anjali, Srisaimaneesh Yalamanchili, Alexander R. Davis, and Adam W. Barb. "Expression and Display of Glycoengineered Antibodies and Antibody Fragments with an Engineered Yeast Strain." Antibodies 10, no. 4 (September 29, 2021): 38. http://dx.doi.org/10.3390/antib10040038.

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Interactions with cell surface receptors enhance the therapeutic properties of many important antibodies, including the low-affinity Fc γ Receptors (FcγRs). These interactions require proper processing of the immunoglobulin G Fc N-glycan, and eliminating the N-glycan abolishes binding, restricting antibody production to mammalian expression platforms. Yeasts, for example, generate extensively mannosylated N-glycans that are unsuitable for therapeutics. However, Fc with a specifically truncated N-glycan still engages receptors with considerable affinity. Here we describe the creation and applications of a novel Saccharomyces cerevisiae strain that specifically modifies the IgG1 Fc domain with an N-glycan consisting of a single N-acetylglucosamine residue. This strain displayed glycoengineered Fc on its surface for screening yeast surface display libraries and also served as an alternative platform to produce glycoengineered Rituximab. An IgG-specific endoglycosidase (EndoS2) truncates the IgG1 Fc N-glycan. EndoS2 was targeted to the yeast ER using the signal peptide from the yeast protein disulfide isomerase (PDI) and a yeast ER retention signal (HDEL). Furthermore, >99% of the yeast expressed Rituximab displayed the truncated glycoform as determined by SDS-PAGE and ESI-MS analyses. Lastly, the yeast expressed Rituximab engaged the FcγRIIIa with the expected affinity (KD = 2.0 ± 0.5 μM) and bound CD20 on Raji B cells.

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Vialás, Vital, Palani Perumal, Dolores Gutierrez, Pilar Ximénez-Embún, César Nombela, Concha Gil, and W. LaJean Chaffin. "Cell surface shaving of Candida albicans biofilms, hyphae, and yeast form cells." PROTEOMICS 12, no. 14 (August 2012): 2331–39. http://dx.doi.org/10.1002/pmic.201100588.

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45

Zou, Wen, Mitsuyoshi Ueda, and Atsuo Tanaka. "Genetically Controlled Self-Aggregation of Cell-Surface-Engineered Yeast Responding to Glucose Concentration." Applied and Environmental Microbiology 67, no. 5 (May 1, 2001): 2083–87. http://dx.doi.org/10.1128/aem.67.5.2083-2087.2001.

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ABSTRACT We constructed an arming (cell-surface-engineered) yeast displaying two types of agglutinin (modified a-agglutinin and α-agglutinin) on the cell surface, with agglutination being independent of both mating type and pheromones. The modified a-agglutinin was artificially prepared by the fusion of the genes encoding Aga1p and Aga2p. The modified a-agglutinin could induce agglutination of cells displaying Agα1p (α-agglutinin). The upstream region of the isocitrate lyase gene of Candida tropicalis (UPR-ICL), active at a low glucose concentration, was used as the promoter to express the modified a-agglutinin- and α-agglutinin-encoding genes. The arming yeast displaying both agglutinins agglutinated and sedimented in response to decreased glucose concentration. When the glucose concentration was high, the arming yeast grew normally. In the late log phase, when the glucose concentration became very low, agglutination occurred suddenly and drastically and yeast cells sedimented completely. Sedimentation was confirmed by weighing the aggregated cells after filtration of the broth. Strains in which aggregation can be genetically controlled can be used in industrial processes in which the separation of yeast cells from the supernatant is necessary.

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Klotz, Stephen A., Nicole Bradley, and Peter N. Lipke. "Blocking Serum Amyloid-P Component from Binding to Macrophages and Augmenting Fungal Functional Amyloid Increases Macrophage Phagocytosis of Candida albicans." Pathogens 11, no. 9 (September 1, 2022): 1000. http://dx.doi.org/10.3390/pathogens11091000.

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Candida-macrophage interactions are important immune defense responses associated with disseminated and deep-seated candidiasis in humans. Cells of Candida spp. express functional amyloids on their surfaces during the pathogenesis of disseminated candidiasis. These amyloids become decorated with serum amyloid P-component (SAP) that binds to Candida cells and macrophages and downregulates the cellular and cytokine response to the fungi. In this report, further characterization of the interactions of SAP and fungal functional amyloid are demonstrated. Blocking the binding of SAP to macrophage FcγR1 receptors increases phagocytosis of yeast cells; seeding a pro-amyloid-forming peptide on the yeast cell surface also increases phagocytosis of yeasts by macrophages; and, lastly, miridesap, a small palindromic molecule, prevents binding of SAP to yeasts and removes SAP that is bound to C. albicans thus, potentially increasing phagocytosis of yeasts by macrophages. Some, or all, of these interventions may be useful in boosting the host immune response to disseminated candidiasis.

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Breinig, Frank, Björn Diehl, Sabrina Rau, Christian Zimmer, Helmut Schwab, and Manfred J. Schmitt. "Cell Surface Expression of Bacterial Esterase A by Saccharomyces cerevisiae and Its Enhancement by Constitutive Activation of the Cellular Unfolded Protein Response." Applied and Environmental Microbiology 72, no. 11 (September 15, 2006): 7140–47. http://dx.doi.org/10.1128/aem.00503-06.

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ABSTRACT Yeast cell surface display is a powerful tool for expression and immobilization of biocatalytically active proteins on a unicellular eukaryote. Here bacterial carboxylesterase EstA from Burkholderia gladioli was covalently anchored into the cell wall of Saccharomyces cerevisiae by in-frame fusion to the endogenous yeast proteins Kre1p, Cwp2p, and Flo1p. When p-nitrophenyl acetate was used as a substrate, the esterase specific activities of yeast expressing the protein fusions were 103 mU mg−1 protein for Kre1/EstA/Cwp2p and 72 mU mg−1 protein for Kre1/EstA/Flo1p. In vivo cell wall targeting was confirmed by esterase solubilization after laminarinase treatment and immunofluorescence microscopy. EstA expression resulted in cell wall-associated esterase activities of 2.72 U mg−1 protein for Kre1/EstA/Cwp2p and 1.27 U mg−1 protein for Kre1/EstA/Flo1p. Furthermore, esterase display on the yeast cell surface enabled the cells to effectively grow on the esterase-dependent carbon source glycerol triacetate (Triacetin). In the case of Kre1/EstA/Flo1p, in vivo maturation within the yeast secretory pathway and final incorporation into the wall were further enhanced when there was constitutive activation of the unfolded protein response pathway. Our results demonstrate that esterase cell surface display in yeast, which, as shown here, is remarkably more effective than EstA surface display in Escherichia coli, can be further optimized by activating the protein folding machinery in the eukaryotic secretion pathway.

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48

Tsygankov, Miklhail A., Andrey M. Rumyantsev, Anastasiya S. Makeeva та Marina V. Padkina. "Comparasion of the effectiveness of anchor proteins ScAGα1p, KpCW51p, KpCW61p for surface display in yeast <i>Komagataella phaffii</i>". Ecological genetics 20, № 4 (24 грудня 2022): 359–71. http://dx.doi.org/10.17816/ecogen112509.

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BACKGROUND: Yeast display is an effective technology for exposure target proteins to the cell surface by fusing them with cell wall proteins. This technique, among other things, makes it possible to obtain vaccine preparations based on yeast by exposing antigen proteins on their cell surface. Finding and selecting proteins that allow effective exposure of target proteins on the surface of yeast cells is an urgent task. AIM: The aim of this work was to evaluate the efficiency of cell wall proteins ScAG1p, KpCW51p, KpCW61p for displaying the reporter protein on the Komagataella phaffii cell surface, including the study of several variants of the ScAG1 gene coding sequence. MATERIALS AND METHODS: The studied gene sequences were cloned under the control of the AOX1 gene promoter in the same reading frame as the eGFP reporter protein gene and integrated into the genome of the K. Phaffii yeast strain X-33. RESULTS: Cytoimmunochemical analysis and confocal microscopy of strains displaying the eGFP protein on their surface under conditions of induction of the AOX1 gene promoter made it possible to identify the most effective anchor protein. The best efficiency was demonstrated for the sequence of the ScAG1 gene without the native 3' non-coding region. CONCLUSIONS: The plasmids obtained in the work will make it possible to obtain a yeast strain K. phaffii that effectively exposure proteins, including antigens, on its surface, which can be used as a vaccine preparation.

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49

Le, Phuc H., Duy H. K. Nguyen, Arturo Aburto Medina, Denver P. Linklater, Christian Loebbe, Russell J. Crawford, Shane MacLaughlin, and Elena P. Ivanova. "Surface Architecture Influences the Rigidity of Candida albicans Cells." Nanomaterials 12, no. 3 (February 7, 2022): 567. http://dx.doi.org/10.3390/nano12030567.

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Atomic force microscopy (AFM) was used to investigate the morphology and rigidity of the opportunistic pathogenic yeast, Candida albicans ATCC 10231, during its attachment to surfaces of three levels of nanoscale surface roughness. Non-polished titanium (npTi), polished titanium (pTi), and glass with respective average surface roughness (Sa) values of 389 nm, 14 nm, and 2 nm, kurtosis (Skur) values of 4, 16, and 4, and skewness (Sskw) values of 1, 4, and 1 were used as representative examples of each type of nanoarchitecture. Thus, npTi and glass surfaces exhibited similar Sskw and Skur values but highly disparate Sa. C. albicans cells that had attached to the pTi surfaces exhibited a twofold increase in rigidity of 364 kPa compared to those yeast cells attached to the surfaces of npTi (164 kPa) and glass (185 kPa). The increased rigidity of the C. albicans cells on pTi was accompanied by a distinct round morphology, condensed F-actin distribution, lack of cortical actin patches, and the negligible production of cell-associated polymeric substances; however, an elevated production of loose extracellular polymeric substances (EPS) was observed. The differences in the physical response of C. albicans cells attached to the three surfaces suggested that the surface nanoarchitecture (characterized by skewness and kurtosis), rather than average surface roughness, could directly influence the rigidity of the C. albicans cells. This work contributes to the next-generation design of antifungal surfaces by exploiting surface architecture to control the extent of biofilm formation undertaken by yeast pathogens and highlights the importance of performing a detailed surface roughness characterization in order to identify and discriminate between the surface characteristics that may influence the extent of cell attachment and the subsequent behavior of the attached cells.

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Buck, James W., and John H. Andrews. "Localized, Positive Charge Mediates Adhesion ofRhodosporidium toruloides to Barley Leaves and Polystyrene." Applied and Environmental Microbiology 65, no. 5 (May 1, 1999): 2179–83. http://dx.doi.org/10.1128/aem.65.5.2179-2183.1999.

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ABSTRACT The physicochemical forces that mediate attachment of yeasts to the phylloplane are unknown. Cell surface charge and hydrophobicity and adhesion to polystyrene, glass, and barley were assessed for wild-typeRhodosporidium toruloides and attachment-minus (Att−) mutants. Cells were grown under conditions promoting (excess carbon) or not promoting (excess nitrogen) capsule production. Hydrophobicity was measured by adhesion to xylenes, and surface charge characteristics were assessed by attachment to either DEAE (positive)- or carboxymethyl (CM) (negative)-Sephadex ion-exchange beads. Hydrophobicity and adhesiveness of nonencapsulated, wild-typeR. toruloides decreased from mid-log to late stationary phase. Encapsulated wild-type R. toruloides cells were more hydrophobic and more adhesive than nonencapsulated cells. However, two encapsulated Att− mutants were more hydrophobic than the wild type and levels of adhesion of R. toruloides were similar on polystyrene and less hydrophobic glass surfaces. Adhesion of wild-type yeast to barley and polystyrene was correlated with attachment to CM-Sephadex beads, indicating a positive cell surface charge. Sixteen Att− mutants did not exhibit a positive cell surface charge, and wild-type yeast cells that did not attach to CM-Sephadex did not adhere to either polystyrene or barley. Wild-typeR. toruloides attached to CM-Sephadex beads by the poles of the cells, indicating a localization of positive charge which was also visualized with India ink. We conclude that localized, positive charge, and not hydrophobic interactions, mediates attachment of R. toruloides to barley leaves.

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