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Catherine Madzak - One of the best experts on this subject based on the ideXlab platform.

  • Heterologous expression of histidine acid phytase from pantoea sp. 3.5.1 in Yarrowia lipolytica
    BioNanoScience, 2019
    Co-Authors: Aliya D. Suleimanova, Catherine Madzak, Daria S. Troshagina, Margarita R. Sharipova
    Abstract:

    The dimorphic yeasts Yarrowia lipolytica are used as effective expression system and are characterized by a high level of production of heterologous protein. In this work, we aimed to clone and express Pantoea sp. 3.5.1 agpP phytase gene in Yarrowia lipolytica. Genetic constructs containing the native phytase gene (agpP) under the control of the strong hybrid promoter hp4d and the signal peptide of the alkaline extracellular protease XPR2 gene, as well as the optimized gene (agpP-opt) of phytase under the control of two signal peptides—bacterial and yeast were obtained. Recombinant Y. lipolytica strains with integrated bacterial phytase genes were obtained and expression of phytase was analyzed.

  • Effect of different substrates on heterologous production of a fungal laccase in a recombinant strain of Yarrowia lipolytica
    2015
    Co-Authors: Catherine Madzak, M Moradi, Farshad Darvishi, Claude Jolivalt
    Abstract:

    Effect of different substrates on heterologous production of a fungal laccase in a recombinant strain of Yarrowia lipolytica. 16. International and Iranian Congress of Microbiology

  • Designing artificial merozoites for a potential blood stage malaria vaccine by surface display of multiple Plasmodium antigens on Yarrowia yeast cells
    2015
    Co-Authors: Catherine Madzak, Bart Faber, Roberto Rodriguez Garcia
    Abstract:

    Designing artificial merozoites for a potential blood stage malaria vaccine by surface display of multiple Plasmodium antigens on Yarrowia yeast cells. 8. European Recombinant Protein Production Meeting (RPP8)

  • Heterologous Protein Expression and Secretion in Yarrowia lipolytica
    Yarrowia lipolytica, 2013
    Co-Authors: Catherine Madzak, Jean-marie Beckerich
    Abstract:

    Heterologous protein production constitutes an important research field, having both academic and commercial applications. The use of yeasts as host systems took advantage of the combination of their easy manipulation and high growth capacity, with their eukaryotic subcellular organisation allowing post-translational processing. Among available yeast host systems, Yarrowia lipolytica (Yarrowia) appears as one of the most attractive. This non-conventional dimorphic yeast has been distinguished due to its remarkable regularity of performance in the efficient secretion of various heterologous proteins. We will present in this chapter Yarrowia’s characteristics regarding heterologous protein production and describe the genetic and molecular tools available. Recent developments of the Yarrowia expression system, such as surface display vectors, engineered strains and high-throughput screening processes, will be reviewed. A comprehensive survey of the literature allowed us to list more than 100 heterologous proteins, from more than 60 species, successfully produced until now in this yeast. This amount of data sometimes offers a comparison with other yeast host systems and globally demonstrates the reliability and versatility of Yarrowia as host for heterologous production.

  • Tunable nano-oleosomes derived from engineered Yarrowia lipolytica
    Biotechnology and Bioengineering, 2013
    Co-Authors: Zhenlin Han, Catherine Madzak
    Abstract:

    Oleosomes are discrete organelles filled with neutral lipids surrounded by a protein-embedded phospholipid monolayer. Their simple yet robust structure, as well as their amenability to biological, chemical, and physical processing, can be exploited for various biotechnology applications. In this study, we report facile biosynthesis of functionalized oleosomes within oleaginous yeast Yarrowia lipolytica, through expression of oleosin fusion proteins. By fusing a cDNA clone of a sesame oleosin with either the coding sequence of a red fluorescent protein mCherry or a cellulosomal scaffolding protein cohesin from Clostridium cellulolyticum, these oleosin-fusion proteins were efficiently expressed and specifically targeted to and anchored on the surface of the oleosomes within the Y. lipolytica cells. The engineered oleosomes can be easily separated from the Y. lipolytica cell extract via floating centrifugation and both mCherry and cohesin domains are shown to be functional. Upon sonication, the engineered Yarrowia oleosomes exhibit a mean diameter of 200300nm and are found to be highly stable. The feasibility of co-displaying multiple proteins on the Yarrowia oleosomes was demonstrated by incubating cohesin-displaying oleosomes with different dockerin-fusion proteins. Based on this strategy, engineered oleosomes with both cell-targeting and reporting activities were created and shown to be functional. Taken together, the Yarrowia oleosome surface display system in which oleosin serves as an efficient membrane anchor motif shows great promise as a simple platform for creating tunable nanoparticles. Biotechnol. Bioeng. 2013; 110: 702710. (c) 2012 Wiley Periodicals, Inc.

Jean-marie Beckerich - One of the best experts on this subject based on the ideXlab platform.

Zhenlin Han - One of the best experts on this subject based on the ideXlab platform.

  • Tunable nano-oleosomes derived from engineered Yarrowia lipolytica
    Biotechnology and Bioengineering, 2013
    Co-Authors: Zhenlin Han, Catherine Madzak
    Abstract:

    Oleosomes are discrete organelles filled with neutral lipids surrounded by a protein-embedded phospholipid monolayer. Their simple yet robust structure, as well as their amenability to biological, chemical, and physical processing, can be exploited for various biotechnology applications. In this study, we report facile biosynthesis of functionalized oleosomes within oleaginous yeast Yarrowia lipolytica, through expression of oleosin fusion proteins. By fusing a cDNA clone of a sesame oleosin with either the coding sequence of a red fluorescent protein mCherry or a cellulosomal scaffolding protein cohesin from Clostridium cellulolyticum, these oleosin-fusion proteins were efficiently expressed and specifically targeted to and anchored on the surface of the oleosomes within the Y. lipolytica cells. The engineered oleosomes can be easily separated from the Y. lipolytica cell extract via floating centrifugation and both mCherry and cohesin domains are shown to be functional. Upon sonication, the engineered Yarrowia oleosomes exhibit a mean diameter of 200300nm and are found to be highly stable. The feasibility of co-displaying multiple proteins on the Yarrowia oleosomes was demonstrated by incubating cohesin-displaying oleosomes with different dockerin-fusion proteins. Based on this strategy, engineered oleosomes with both cell-targeting and reporting activities were created and shown to be functional. Taken together, the Yarrowia oleosome surface display system in which oleosin serves as an efficient membrane anchor motif shows great promise as a simple platform for creating tunable nanoparticles. Biotechnol. Bioeng. 2013; 110: 702710. (c) 2012 Wiley Periodicals, Inc.

  • Tunable nano-oleosomes derived from engineered Yarrowia lipolytica.
    Biotechnology and bioengineering, 2012
    Co-Authors: Zhenlin Han, Catherine Madzak
    Abstract:

    Oleosomes are discrete organelles filled with neutral lipids surrounded by a protein-embedded phospholipid monolayer. Their simple yet robust structure, as well as their amenability to biological, chemical, and physical processing, can be exploited for various biotechnology applications. In this study, we report facile biosynthesis of functionalized oleosomes within oleaginous yeast Yarrowia lipolytica, through expression of oleosin fusion proteins. By fusing a cDNA clone of a sesame oleosin with either the coding sequence of a red fluorescent protein mCherry or a cellulosomal scaffolding protein cohesin from Clostridium cellulolyticum, these oleosin-fusion proteins were efficiently expressed and specifically targeted to and anchored on the surface of the oleosomes within the Y. lipolytica cells. The engineered oleosomes can be easily separated from the Y. lipolytica cell extract via floating centrifugation and both mCherry and cohesin domains are shown to be functional. Upon sonication, the engineered Yarrowia oleosomes exhibit a mean diameter of 200-300 nm and are found to be highly stable. The feasibility of co-displaying multiple proteins on the Yarrowia oleosomes was demonstrated by incubating cohesin-displaying oleosomes with different dockerin-fusion proteins. Based on this strategy, engineered oleosomes with both cell-targeting and reporting activities were created and shown to be functional. Taken together, the Yarrowia oleosome surface display system in which oleosin serves as an efficient membrane anchor motif shows great promise as a simple platform for creating tunable nanoparticles.

Cletus P. Kurtzman - One of the best experts on this subject based on the ideXlab platform.

  • a survey of yeast from the Yarrowia clade for lipid production in dilute acid pretreated lignocellulosic biomass hydrolysate
    Applied Microbiology and Biotechnology, 2017
    Co-Authors: Josh Quarterman, Cletus P. Kurtzman, Patricia J Slininger, Stephanie R Thompson, Bruce S Dien
    Abstract:

    Yarrowia lipolytica is an oleaginous yeast species that has attracted attention as a model organism for synthesis of single cell oil. Among over 50 isolates of Y. lipolytica identified, only a few of the strains have been studied extensively. Furthermore, 12 other yeast species were recently assigned to the Yarrowia clade, and most are not well characterized in terms of cell growth and lipid accumulation, especially in industrially relevant conditions. In the present study, we investigated biomass and lipid production by 57 yeast isolates, representing all 13 species in the Yarrowia clade, on a non-detoxified dilute acid-pretreated switchgrass hydrolysate under highly aerobic conditions. The objective was to compare yeast physiology during growth in an abundant, low-cost biomass feedstock and to expand diversity of genetically tractable, oleaginous yeasts available for lipid research. Screening of 45 Y. lipolytica isolates demonstrated considerable variation within the species in terms of lipid accumulation (min = 0.1 g/L; max = 5.1 g/L; mean = 2.3 g/L); three strains (NRRL YB-420, YB-419, and YB-392) were especially promising for cellulosic biomass conversion with average improvements of 43, 57, and 64%, respectively, in final lipid titer as compared to control strain W29. Subsequently, evaluation of strains from 13 distinct species in the Yarrowia clade identified Candida phangngensis PT1-17 as the top lipid producer with a maximum titer of 9.8 g/L lipid, which was over twofold higher than the second-best species in the clade (Candida hollandica NRRL Y-48254). A small set of the most promising strains from the screenings was further characterized to evaluate inhibitor tolerance, lipid production kinetics, and fatty acid distribution. We expect that the results of this study will pave the way for new biotechnological applications involving previously overlooked and under-characterized strains within the Yarrowia clade.

  • Yarrowia van der Walt & von Arx (1980)
    The Yeasts, 2011
    Co-Authors: Cletus P. Kurtzman
    Abstract:

    Publisher Summary This chapter studies the genus Yarrowia. Aasexual reproduction is by multilateral budding on a narrow base. Arthroconidia may also be formed. Budded cells are spherical, ellipsoidal, or often elongate. True hyphae and pseudohyphae are formed; true hyphae have septa with a single central micropore. In sexual reproduction it is found that the asci are unconjugated and generally arise from diploid hyphae. The asci, which slowly deliquesce, form one to four ascospores that may be spherical, hat-shaped, hemispheroidal, or somewhat angular. The single described species is heterothallic. The chapter also discusses physiology/biochemistry and phylogenetic placement of the genus in which sugars are not fermented, nitrate is not assimilated, and lipases and proteases are produced. Coenzyme Q-9 is formed and the diazonium blue B reaction is negative. The type species taken is Yarrowia lipolytica. Yarrowia lipolytica, the only described species of the genus Yarrowia, is widespread in nature and has considerable industrial utility as well as importance to the food and medical fields.

  • Characterization of Yarrowia lipolytica and related species for citric acid production from glycerol
    Enzyme and Microbial Technology, 2007
    Co-Authors: William E. Levinson, Cletus P. Kurtzman, Tsung Min Kuo
    Abstract:

    Abstract Twenty-seven Yarrowia lipolytica strains as well as five strains from three other species of the Yarrowia clade ( Aciculoconidium aculeatum , Candida hispaniensis and Candida bentonensis ) were screened for citric acid production with pure glycerol as the carbon source. The cultures were grown under nitrogen-limited conditions. None of the non- Yarrowia strains produced citric acid, although they were able to grow on glycerol. All of the Y. lipolytica strains were able to produce citric acid in varying concentrations and, under the screening conditions used, the yields obtained formed a continuum from very low to among the highest reported for this substrate. The highest yielding strain, Y. lipolytica NRRL YB-423, produced 21.6 g/L citric acid from 40 g/L glycerol (54% yield). The citric acid to isocitric acid ratio produced by this strain in the initial screen was 11.3, while most of the strains produced ratios of between 2 and 6. Further work on medium optimization with this strain showed that the optimum carbon to nitrogen ratio for the rate of citric acid production was 172 while the best combination of rate and yield was obtained at a C/N ratio of 343. The citric acid to isocitric acid ratios produced reached an optimum at C/N ratios of 343–686.

João A. P. Coutinho - One of the best experts on this subject based on the ideXlab platform.

  • Deposition of Yarrowia lipolytica on plasma prepared teflonlike thin films
    Surface Engineering, 2008
    Co-Authors: Marián Lehocký, Priscilla Filomena Fonseca Amaral, Maria Alice Zarur Coelho, Ana Barros-timmons, Pavel Sťahel, João A. P. Coutinho
    Abstract:

    The adhesion of Yarrowia lipolytica to teflonlike thin films deposited by plasma on polycarbonate substrates was investigated through a series of tests in order to develop a substrate for strong and selective adhesion of Yarrowia lipolytica cells. Teflonlike thin films were prepared using atmospheric pressure surface barrier discharge with mixtures of octafluorocyclobutane (C4F8) and nitrogen as plasma gas. A variety of plasma gas feedrates and different deposition times were studied. The films were characterised by Fourier transform infrared and contact angle measurements using the sessile drop technique. Total surface energy and its components were calculated using the acid base theory. Attachment of the yeast cells was assessed by optical and scanning electron microscopy. The optimal deposition conditions for cell adhesion were determined using standard adhesion tests.

  • Preparation and characterization of organosilicon thin films for selective adhesion of Yarrowia lipolytica yeast cells
    Journal of Chemical Technology & Biotechnology, 2007
    Co-Authors: Marián Lehocký, Priscilla Filomena Fonseca Amaral, Pavel Stahel, Maria Alice Zarur Coelho, Ana Barros-timmons, João A. P. Coutinho
    Abstract:

    Preparation and characterization of organosilicon thin films for selective adhesion of Yarrowia lipolytica yeast cells

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