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Hak Jun Kim - One of the best experts on this subject based on the ideXlab platform.

  • structure based characterization and antifreeze properties of a hyperactive ice binding protein from the antarctic bacterium flavobacterium frigoris ps1
    Acta Crystallographica Section D-biological Crystallography, 2014
    Co-Authors: Soonjong Kim, Hak Jun Kim, Jun Hyuck Lee
    Abstract:

    Ice-binding proteins (IBPs) inhibit ice growth through direct interaction with ice crystals to permit the survival of polar organisms in extremely cold environments. FfIBP is an ice-binding protein encoded by the Antarctic bacterium Flavobacterium frigoris PS1. The X-ray crystal structure of FfIBP was determined to 2.1 A resolution to gain insight into its ice-binding mechanism. The refined structure of FfIBP shows an intramolecular disulfide bond, and analytical ultracentrifugation and analytical size-exclusion chromatography show that it behaves as a monomer in solution. Sequence alignments and structural comparisons of IBPs allowed two groups of IBPs to be defined, depending on sequence differences between the α2 and α4 loop regions and the presence of the disulfide bond. Although FfIBP closely resembles Leucosporidium (recently re-classified as Glaciozyma) IBP (LeIBP) in its amino-acid sequence, the thermal hysteresis (TH) activity of FfIBP appears to be tenfold higher than that of LeIBP. A comparison of the FfIBP and LeIBP structures reveals that FfIBP has different ice-binding residues as well as a greater surface area in the ice-binding site. Notably, the ice-binding site of FfIBP is composed of a T-A/G-X-T/N motif, which is similar to the ice-binding residues of hyperactive antifreeze proteins. Thus, it is proposed that the difference in TH activity between FfIBP and LeIBP may arise from the amino-acid composition of the ice-binding site, which correlates with differences in affinity and surface complementarity to the ice crystal. In conclusion, this study provides a molecular basis for understanding the antifreeze mechanism of FfIBP and provides new insights into the reasons for the higher TH activity of FfIBP compared with LeIBP.

  • optimization of the pilot scale production of an ice binding protein by fed batch culture of pichia pastoris
    Applied Microbiology and Biotechnology, 2013
    Co-Authors: Jun Hyuck Lee, Sung Gu Lee, Eunjung Kim, Jongchan Park, Yonghoe Choe, Se Jong Han, Hak Jun Kim
    Abstract:

    Ice-binding proteins (IBPs) can bind to the ice crystal and inhibit its growth. Because this property of IBPs can increase the freeze–thaw survival of cells, IBPs have attracted the attention from industries for their potential use in biotechnological applications. However, their use was largely hampered by the lack of the large-scale recombinant production system. In this study, the codon-optimized IBP from Leucosporidium sp. (LeIBP) was constructed and subjected to high-level expression in methylotrophic Pichia pastoris system. In a laboratory-scale fermentation (7 L), the optimal induction temperature and pH were determined to be 25 °C and 6.0, respectively. Further, employing glycerol fed-batch phase prior to methanol induction phase enhanced the production of recombinant LelBP (rLeIBP) by ∼100 mg/l. The total amount of secreted proteins at these conditions (25 °C, pH 6.0, and glycerol fed-batch phase) was ∼443 mg/l, 60 % of which was rLeIBP, yielding ∼272 mg/l. In the pilot-scale fermentation (700 L) under the same conditions, the yield of rLeIBP was 300 mg/l. To our best knowledge, this result reports the highest production yield of the recombinant IBP. More importantly, the rLeIBP secreted into culture media was stable and active for 6 days of fermentation. The thermal hysteresis (TH) activity of rLeIBP was about 0.42 °C, which is almost the same to those reported previously. The availability of large quantities of rLeIBP may accelerate further application studies.

  • characterization of the ice binding protein from arctic yeast Leucosporidium sp ay30
    Cryobiology, 2012
    Co-Authors: Kyoung Sun Park, Jun Hyuck Lee, Seungil Park, Eunjung Kim, Soonjong Kim, Sungho Kang, Hak Jun Kim
    Abstract:

    Abstract Previously, we reported the ice-binding protein (LeIBP) from the Arctic yeast Leucosporidium sp. AY30. In this study we provide physicochemical characterization of this IBP, which belongs to a class of IBPs that exhibited no significant similarity in primary structure to other known antifreeze proteins (AFPs). We compared native, glycosylated and non-glycosylated recombinant LeIBPs. Interestingly, size-exclusion chromatography and analytical ultracentrifugation revealed that LeIBP self-associates with a reversible dimer with K d values in the range 3.45–7.24 × 10 −6  M. Circular dichroism (CD) spectra showed that LeIBP, glycosylated or non-glycosylated, is predominantly composed of β-strand secondary structural elements (54.6%), similar to other β-helical antifreeze proteins (AFPs). In thermal hysteresis (TH) activity measurements, native LeIBP was twice more active (0.87 °C at 15 mg/mL) than that of the recombinant IBPs (0.43–0.42 °C at 10.8 mg/mL). This discrepancy is probably due to uncharacterized enhancing factors carried over during ice affinity purification, because glycosylated and non-glycosylated recombinant proteins displayed similarly low activity. Ice recrystallization inhibition (RI) activities of the native and recombinant LeIBPs were comparable. Measurements of CD, TH activity, and RI showed that glycosylation does not cause structural changes and is not required for function. An ice-etching experiment using green fluorescent protein-tagged IBP revealed that LeIBP binds, just as hyperactive AFPs, to both basal and pyramidal prism planes of the ice crystal. Taken together, our results indicate that LeIBP, structurally similar to hyperactive AFPs, is moderately active and that a reversible dimer has no effect on its activity.

  • cryopreservative effects of the recombinant ice binding protein from the arctic yeast Leucosporidium sp on red blood cells
    Applied Biochemistry and Biotechnology, 2012
    Co-Authors: Sung Gu Lee, Jun Hyuck Lee, Sungho Kang, Hye Yeon Koh, Hak Jun Kim
    Abstract:

    Antifreeze proteins (AFPs) have important functions in many freeze-tolerant organisms. The proteins non-colligatively lower the freezing point and functionally inhibit ice recrystallization in frozen solutions. In our previous studies, we found that the Arctic yeast Leucosporidium sp. produces an AFP (LeIBP), and that the protein could be successfully produced in Pichia expression system. The present study showed that recombinant LeIBP possesses the ability to reduce the damage induced to red blood cells (RBCs) by freeze thawing. In addition to 40 % glycerol, both 0.4 and 0.8 mg/ml LeIBPs significantly reduced freeze–thaw-induced hemolysis at either rapid- (45 °C) or slow-warming (22 °C) temperatures. Post-thaw cell counts of the cryopreserved RBCs were dramatically enhanced, in particular, in 0.8 mg/ml LeIBP. Interestingly, the cryopreserved cells in the presence of LeIBP showed preserved cell size distribution. These results indicate that the ability of LeIBP to inhibit ice recrystallization helps the RBCs avoid critically damaging electrolyte concentrations, which are known as solution effects. Considering all these data, LeIBP can be thought of as a key component in improving RBC cryopreservation efficiency.

  • structural basis for antifreeze activity of ice binding protein from arctic yeast
    Journal of Biological Chemistry, 2012
    Co-Authors: Jun Hyuck Lee, Kyoung Sun Park, Ae Kyung Park, Young Min Chi, Sang Hyun Moh, Hak Jun Kim
    Abstract:

    Arctic yeast Leucosporidium sp. produces a glycosylated ice-binding protein (LeIBP) with a molecular mass of ∼25 kDa, which can lower the freezing point below the melting point once it binds to ice. LeIBP is a member of a large class of ice-binding proteins, the structures of which are unknown. Here, we report the crystal structures of non-glycosylated LeIBP and glycosylated LeIBP at 1.57- and 2.43-A resolution, respectively. Structural analysis of the LeIBPs revealed a dimeric right-handed β-helix fold, which is composed of three parts: a large coiled structural domain, a long helix region (residues 96-115 form a long α-helix that packs along one face of the β-helix), and a C-terminal hydrophobic loop region ((243)PFVPAPEVV(251)). Unexpectedly, the C-terminal hydrophobic loop region has an extended conformation pointing away from the body of the coiled structural domain and forms intertwined dimer interactions. In addition, structural analysis of glycosylated LeIBP with sugar moieties attached to Asn(185) provides a basis for interpreting previous biochemical analyses as well as the increased stability and secretion of glycosylated LeIBP. We also determined that the aligned Thr/Ser/Ala residues are critical for ice binding within the B face of LeIBP using site-directed mutagenesis. Although LeIBP has a common β-helical fold similar to that of canonical hyperactive antifreeze proteins, the ice-binding site is more complex and does not have a simple ice-binding motif. In conclusion, we could identify the ice-binding site of LeIBP and discuss differences in the ice-binding modes compared with other known antifreeze proteins and ice-binding proteins.

Jun Hyuck Lee - One of the best experts on this subject based on the ideXlab platform.

  • the protective effect of Leucosporidium derived ice binding protein leibp on bovine oocytes and embryos during vitrification
    Theriogenology, 2020
    Co-Authors: Wusheng Sun, Hoon Jang, Hyo J Kwon, Ki Young Kim, Soo Bin Ahn, Seongsoo Hwang, Sung Gu Lee, Jun Hyuck Lee, Insul Hwang, Jeongwoong Lee
    Abstract:

    Ice-binding proteins (IBPs) facilitate organism survival under extreme conditions by inhibiting thermal hysteresis and ice recrystallization. IBPs have been widely used as cryoprotectants to cryopreserve mammalian gametes and embryos. In the present study, we evaluated the protective effects of an Arctic yeast, Leucosporidium sp. AY30 derived ice-binding protein (LeIBP), on the vitrification of bovine metaphase II (MII) oocytes and embryos. When oocytes and embryos were frozen using the two-step vitrification method, the survival rate was significantly increased in the presence of LeIBP. The LeIBP supplementation decreased the levels of intracellular reactive oxygen species (ROS) and enhanced mitochondrial functions in the vitrified-warmed oocytes. Furthermore, LeIBP improved the developmental potential and suppressed apoptosis of the embryos derived from vitrified-warmed oocytes. Collectively, these data indicate that LeIBP can be used as a promising cryoprotectant to prevent cryoinjury during vitrification in bovine oocytes.

  • effects of Leucosporidium derived ice binding protein leibp on bull semen cryopreservation
    Veterinary Medicine and Science, 2020
    Co-Authors: Hoon Jang, Hyo J Kwon, Seongsoo Hwang, Sung Gu Lee, Jun Hyuck Lee, Wu S Sun, In S Hwang, Sung Woo Kim, Jeong W Lee
    Abstract:

    We examined the effect of ice-binding protein derived from Leucosporidium (LeIBP) on the cryopreservation of bull semen and compared it with that derived from previously reported Antifreeze Protein III (AFPIII). Six concentrations of LeIBP (10-1  ~ 104  μg/ml) and AFPIII (10-1  ~ 104  μg/ml) were added to the bull semen extender, respectively. Sperm kinematic parameters were measured to examine sperm toxicity and cryopreserved sperm quality. Measures of antioxidant activity such as superoxide dismutase (SOD), reduced glutathione/oxidative glutathione (GSH/GSSG), and total antioxidant capacity (TAC) were analysed to identify the effect of LeIBP on sperm quality. In addition, sperm viability was analysed using a flow cytometer and fluorescence microscope by SYBR14/PI staining. The results showed that the LeIBP groups (0.1, 1 and 10 μg/ml) were less toxic, and the quality of the sperm were dramatically improved in the extenders containing 0.1 μg/ml LeIBP among concentrations of LeIBP and AFPIII. The SOD activity of LeIBP was greater than that of AFPIII and control. In addition, sperm viability was enhanced in the LeIBP-treated group. In summary, LeIBP is a useful cryoprotective adjuvant for bull sperm cryopreservation, and the most efficient concentration of LeIBP is 0.1 μg/ml.

  • Effect of the Antifreeze Protein from the Arctic Yeast Leucosporidium sp. AY30 on Cryopreservation of the Marine Diatom Phaeodactylum tricornutum
    Applied Biochemistry and Biotechnology, 2015
    Co-Authors: Hye Yeon Koh, Jun Hyuck Lee, Se Jong Han, Hyun Park, Sung Gu Lee
    Abstract:

    Antifreeze proteins are a group of proteins that allow organisms to survive in subzero environments. These proteins possess thermal hysteresis and ice recrystallization inhibition activities. In the present study, we demonstrated the efficiency of a recombinant antifreeze protein from the Arctic yeast Leucosporidium sp. AY30, LeIBP, in cryopreservation of the marine diatom Phaeodactylum tricornutum , which is one of the classical model diatoms and has most widely been studied with regard to its ecology, physiology, biochemistry, and molecular biology. P. tricornutum cells were frozen by either a fast or two-step freezing method in freezing medium containing 10 % dimethyl sulfoxide, glycerol, propylene glycol, and ethylene glycol, respectively, with or without LeIBP supplement. When cells were frozen using the two-step freezing method, cell survival was significantly increased and statistically the same as that of unfrozen native cells in the presence of 0.1 mg/ml LeIBP in 10 % propylene glycol or 10 % ethylene glycol at day 11 of post-thaw culture. In the presence of LeIBP, the concentration of chlorophyll a was dramatically increased to 14-, 48-, 1.6-, and 8.8-fold when cells were frozen in freezing medium containing dimethyl sulfoxide (DMSO), glycerol, propylene glycol (PG), and ethylene glycol (EG), respectively. Scanning electron microscopy observations demonstrated that the cells were also successfully preserved and epitheca or hypotheca were not deformed. These results demonstrate that LeIBP was successfully applied to improve cryopreservation of the marine diatom P. tricornutum .

  • structure based characterization and antifreeze properties of a hyperactive ice binding protein from the antarctic bacterium flavobacterium frigoris ps1
    Acta Crystallographica Section D-biological Crystallography, 2014
    Co-Authors: Soonjong Kim, Hak Jun Kim, Jun Hyuck Lee
    Abstract:

    Ice-binding proteins (IBPs) inhibit ice growth through direct interaction with ice crystals to permit the survival of polar organisms in extremely cold environments. FfIBP is an ice-binding protein encoded by the Antarctic bacterium Flavobacterium frigoris PS1. The X-ray crystal structure of FfIBP was determined to 2.1 A resolution to gain insight into its ice-binding mechanism. The refined structure of FfIBP shows an intramolecular disulfide bond, and analytical ultracentrifugation and analytical size-exclusion chromatography show that it behaves as a monomer in solution. Sequence alignments and structural comparisons of IBPs allowed two groups of IBPs to be defined, depending on sequence differences between the α2 and α4 loop regions and the presence of the disulfide bond. Although FfIBP closely resembles Leucosporidium (recently re-classified as Glaciozyma) IBP (LeIBP) in its amino-acid sequence, the thermal hysteresis (TH) activity of FfIBP appears to be tenfold higher than that of LeIBP. A comparison of the FfIBP and LeIBP structures reveals that FfIBP has different ice-binding residues as well as a greater surface area in the ice-binding site. Notably, the ice-binding site of FfIBP is composed of a T-A/G-X-T/N motif, which is similar to the ice-binding residues of hyperactive antifreeze proteins. Thus, it is proposed that the difference in TH activity between FfIBP and LeIBP may arise from the amino-acid composition of the ice-binding site, which correlates with differences in affinity and surface complementarity to the ice crystal. In conclusion, this study provides a molecular basis for understanding the antifreeze mechanism of FfIBP and provides new insights into the reasons for the higher TH activity of FfIBP compared with LeIBP.

  • optimization of the pilot scale production of an ice binding protein by fed batch culture of pichia pastoris
    Applied Microbiology and Biotechnology, 2013
    Co-Authors: Jun Hyuck Lee, Sung Gu Lee, Eunjung Kim, Jongchan Park, Yonghoe Choe, Se Jong Han, Hak Jun Kim
    Abstract:

    Ice-binding proteins (IBPs) can bind to the ice crystal and inhibit its growth. Because this property of IBPs can increase the freeze–thaw survival of cells, IBPs have attracted the attention from industries for their potential use in biotechnological applications. However, their use was largely hampered by the lack of the large-scale recombinant production system. In this study, the codon-optimized IBP from Leucosporidium sp. (LeIBP) was constructed and subjected to high-level expression in methylotrophic Pichia pastoris system. In a laboratory-scale fermentation (7 L), the optimal induction temperature and pH were determined to be 25 °C and 6.0, respectively. Further, employing glycerol fed-batch phase prior to methanol induction phase enhanced the production of recombinant LelBP (rLeIBP) by ∼100 mg/l. The total amount of secreted proteins at these conditions (25 °C, pH 6.0, and glycerol fed-batch phase) was ∼443 mg/l, 60 % of which was rLeIBP, yielding ∼272 mg/l. In the pilot-scale fermentation (700 L) under the same conditions, the yield of rLeIBP was 300 mg/l. To our best knowledge, this result reports the highest production yield of the recombinant IBP. More importantly, the rLeIBP secreted into culture media was stable and active for 6 days of fermentation. The thermal hysteresis (TH) activity of rLeIBP was about 0.42 °C, which is almost the same to those reported previously. The availability of large quantities of rLeIBP may accelerate further application studies.

Lara Duraes Sette - One of the best experts on this subject based on the ideXlab platform.

  • glutaminase free l asparaginase production by Leucosporidium muscorum isolated from antarctic marine sediment
    Preparative Biochemistry & Biotechnology, 2021
    Co-Authors: Rominne Karla Barros Freire, Carlos Miguel Nobrega Mendonca, Rafael Bertelli Ferraro, Ignacio S Moguel, Aldo Tonso, Felipe Rebello Lourenco, Joao H P M Santos, Lara Duraes Sette, Adalberto Pessoa
    Abstract:

    L-asparaginase (ASNase) is an essential drug in the treatment of acute lymphoblastic leukemia (ALL). Commercial bacterial ASNases increase patient survival, but the consequent immunological reactio...

  • statistical experimental design applied to extracellular lipase production by the marine antarctic yeast Leucosporidium scottii crm 728
    Biocatalysis and agricultural biotechnology, 2021
    Co-Authors: Lara Duraes Sette, Alysson Wagner Fernandes Duarte, Rafaella Costa Bonuglisantos, Ana Lucia Ferrarezi Duarte, Eleni Gomes
    Abstract:

    Abstract The search for microbial enzymes from extreme environments may result in the discovery of biomolecules with different properties. In this sense, the aim of this study was to evaluate and optimize the conditions for lipase production by the marine-derived Antarctic yeast Leucosporidium scottii CRM 728 using an experimental design with different sources of carbon and nitrogen. The applied strategy was composed of three steps, including Plackett-Burman, Central Composite Design (CCD), and validation assay. Among nine variables applied, corn steep liquor, olive oil, and soybean oil showed a positive effect in the lipase production, while urea showed a negative effect. The production of lipase increased 9-fold in comparison to the basal activity when olive oil and corn steep liquor were used as nutrient sources. On the other hand, there was an increase of 4.8-fold when soybean oil and corn steep liquor were used as nutrient sources. The highest amounts of lipase were achieved in non-saline conditions and at 20.0 °C. In this study, the lipase production by the marine-derived psychrotolerant yeast L. scottii was for the first time reported, providing new knowledgement in the field of enzymatic production by extremophilic and marine microbial resource.

  • liquid liquid extraction of lipase produced by psychrotrophic yeast Leucosporidium scottii l117 using aqueous two phase systems
    Separation and Purification Technology, 2015
    Co-Authors: Alysson Wagner Fernandes Duarte, Lara Duraes Sette, Adalberto Pessoa, Andre Moreni Lopes, Joao Vitor Dutra Molino
    Abstract:

    Abstract Aqueous two-phase systems (ATPS) have been used in biomolecules separation and as an efficient alternative to traditional purification systems for lipases extraction. Here, we investigated the partitioning and recovery of lipase derived from Leucosporidium scottii L117 using ATPS and aqueous two-phase micellar systems (ATPMS). Thus, we evaluated three ATPS: (i) polyethylene glycol (PEG)/phosphate salts and (ii) PEG/polyacrylic acid (NaPA) in different molecular weights (1500, 4000 and 8000 g/mol). (iii) Triton X-114 (TX-114)/McIlvaine buffer pH 7.0 in different conditions (2.0% (w/w) of TX-114 at 25.0 and 28.0 °C). The PEG/phosphate and PEG/NaPA systems resulted in a great loss of enzymatic activity; thus these systems do not represent viable alternatives for these lipase extraction. The micellar systems yielded the best results for lipase extraction with enzyme activity balances ranging between 84.7% and 113.05%. After optimizing the micellar system by experimental design of the partition coefficient of lipase increased by 10.3-fold (0.75–7.76). Lipase preferentially partitioned into the micelle-rich phase with KLip = 7.76, %RECBot = 93.85% and PF = 1.2 at 25.03 °C, 5.1 pH and 10.38% TX-114 and KLip = 4.77, %RECBot = 73.53% and PF = 1.97 at 28.00 °C, 4.5 pH and 8.0% TX-114, indicating that the ATPMS represents an alternative to purification/extraction of lipase L. scottii L117. A crude lipase extract was also evaluated to define the optimum pH and temperature. Lipase reached optimal activity at 40 °C, and remained stable in pH values ranging from pH 3.0 to 8.0 and temperatures from 20.0 to 45.0 °C, with relative residual lipase activity above 80% after 30 min of incubation.

  • optimization of lipase extraction recovery produced by the psychrotrophic yeast Leucosporidium scottii l117 using aqueous two phase micellar systems
    New Biotechnology, 2014
    Co-Authors: Alysson Wagner Fernandes Duarte, Adalberto Pessoa, Joao Vitor Dutra Molino, Andre Lopes, Lara Duraes Sette
    Abstract:

    Universidade Estadual Paulista, Departamento de Bioquimica e Microbiologia, Instituto de Biociencias de Rio Claro

Sara Filippucci - One of the best experts on this subject based on the ideXlab platform.

  • yeast lipids from cardoon stalks stranded driftwood and olive tree pruning residues as possible extra sources of oils for producing biofuels and biochemicals
    Biotechnology for Biofuels, 2018
    Co-Authors: Giorgia Tasselli, Sara Filippucci, E Borsella, Silvia Dantonio, Mattia Gelosia, Gianluca Cavalaglio, Benedetta Turchetti, Ciro Sannino, Andrea Onofri, Silvio Mastrolitti
    Abstract:

    Some lignocellulosic biomass feedstocks occur in Mediterranean Countries. They are still largely unexploited and cause considerable problems due to the lack of cost-effective harvesting, storage and disposal technologies. Recent studies found that some basidiomycetous yeasts are able to accumulate high amount of intracellular lipids for biorefinery processes (i.e., biofuels and biochemicals). Accordingly, the above biomass feedstocks could be used as carbon sources (after their pre-treatment and hydrolysis) for lipid accumulation by oleaginous yeasts. Cardoon stalks, stranded driftwood and olive tree pruning residues were pre-treated with steam-explosion and enzymatic hydrolysis for releasing free mono- and oligosaccharides. Lipid accumulation tests were performed at two temperatures (20 and 25 °C) using Leucosporidium creatinivorum DBVPG 4794, Naganishia adeliensis DBVPG 5195 and Solicoccozyma terricola DBVPG 5870. S. terricola grown on cardoon stalks at 20 °C exhibited the highest lipid production (13.20 g/l), a lipid yield (28.95%) close to the maximum theoretical value and a lipid composition similar to that found in palm oil. On the contrary, N. adeliensis grown on stranded driftwood and olive tree pruning residues exhibited a lipid composition similar to those of olive and almonds oils. A predictive evaluation of the physical properties of the potential biodiesel obtainable by lipids produced by tested yeast strains has been reported and discussed. Lipids produced by some basidiomycetous yeasts grown on Mediterranean lignocellulosic biomass feedstocks could be used as supplementary sources of oils for producing biofuels and biochemicals.

  • Yeast lipids from cardoon stalks, stranded driftwood and olive tree pruning residues as possible extra sources of oils for producing biofuels and biochemicals
    BMC, 2018
    Co-Authors: Giorgia Tasselli, Sara Filippucci, E Borsella, Mattia Gelosia, Gianluca Cavalaglio, Benedetta Turchetti, Ciro Sannino, Andrea Onofri, Silvia D’antonio, Silvio Mastrolitti
    Abstract:

    Abstract Background Some lignocellulosic biomass feedstocks occur in Mediterranean Countries. They are still largely unexploited and cause considerable problems due to the lack of cost-effective harvesting, storage and disposal technologies. Recent studies found that some basidiomycetous yeasts are able to accumulate high amount of intracellular lipids for biorefinery processes (i.e., biofuels and biochemicals). Accordingly, the above biomass feedstocks could be used as carbon sources (after their pre-treatment and hydrolysis) for lipid accumulation by oleaginous yeasts. Results Cardoon stalks, stranded driftwood and olive tree pruning residues were pre-treated with steam-explosion and enzymatic hydrolysis for releasing free mono- and oligosaccharides. Lipid accumulation tests were performed at two temperatures (20 and 25 °C) using Leucosporidium creatinivorum DBVPG 4794, Naganishia adeliensis DBVPG 5195 and Solicoccozyma terricola DBVPG 5870. S. terricola grown on cardoon stalks at 20 °C exhibited the highest lipid production (13.20 g/l), a lipid yield (28.95%) close to the maximum theoretical value and a lipid composition similar to that found in palm oil. On the contrary, N. adeliensis grown on stranded driftwood and olive tree pruning residues exhibited a lipid composition similar to those of olive and almonds oils. A predictive evaluation of the physical properties of the potential biodiesel obtainable by lipids produced by tested yeast strains has been reported and discussed. Conclusions Lipids produced by some basidiomycetous yeasts grown on Mediterranean lignocellulosic biomass feedstocks could be used as supplementary sources of oils for producing biofuels and biochemicals

  • study of holtermanniella wattica Leucosporidium creatinivorum naganishia adeliensis solicoccozyma aeria and solicoccozyma terricola for their lipogenic aptitude from different carbon sources
    Biotechnology for Biofuels, 2016
    Co-Authors: Sara Filippucci, Giorgia Tasselli, Benedetta Turchetti, Andrea Onofri, Alessandro Scardua, Simone Mauro, Maria Cramarossa, Davide Perini, Luca Forti, Pietro Buzzini
    Abstract:

    The ability of some microorganisms to accumulate lipids is well known; however, only recently the number of studies on microbial lipid biosynthesis for obtaining oleochemical products, namely biofuels and some building blocks for chemistry, is rapidly and spectacularly increased. Since 1990s, some oleaginous yeasts were studied for their ability to accumulate lipids up to 60–70% of their dry weight. Due to the vast array of engineering techniques currently available, the recombinant DNA technology was the main approach followed so far for obtaining lipid-overproducing yeasts, mainly belonging to the Yarrowia lipolytica. However, an alternative approach can be offered by worldwide diversity as source of novel oleaginous yeasts. Lipogenic aptitude of a number of yeast strains has been reviewed, but many of these studies utilized a limited number of species and/or different culture conditions that make impossible the comparison of different results. Accordingly, the lipogenic aptitude inside the yeast world is still far from being fully explored, and finding new oleaginous yeast species can acquire a strategic importance. Holtermanniella wattica, Leucosporidium creatinivorum, Naganishia adeliensis, Solicoccozyma aeria, and Solicoccozyma terricola strains were selected as a result of a large-scale screening on 706 yeasts (both Ascomycota and Basidiomycota). Lipid yields and fatty acid profiles of selected strains were evaluated at 20 and 25 °C on glucose, and on glycerol, xylose, galactose, sucrose, maltose, and cellobiose. A variable fatty acid profile was observed in dependence of both temperature and different carbon sources. On the whole, L. creatinivorum exhibited the highest performances: total lipid yield (YL) >7 g/l on glucose and glycerol, % of intracellular lipids on cell biomass (YL/DW) >70% at 20 °C on glucose, lipid coefficient (YL/Glu) around 20% on glucose, and daily productivity (YL/d) on glucose and sucrose >1.6 g/(l*d). This study provides some meaningful information about the lipogenic ability of some yeast species. Variable lipid yields and fatty acid profiles were observed in dependence of both temperature and different carbon sources. L. creatinivorum exhibited the highest lipogenic performances.

  • MOESM3 of Study of Holtermanniella wattica, Leucosporidium creatinivorum, Naganishia adeliensis, Solicoccozyma aeria, and Solicoccozyma terricola for their lipogenic aptitude from different carbon sources
    2016
    Co-Authors: Sara Filippucci, Giorgia Tasselli, Benedetta Turchetti, Andrea Onofri, Alessandro Scardua, Simone Mauro, Maria Cramarossa, Davide Perini, Luca Forti, Pietro Buzzini
    Abstract:

    Additional file 3: Figure S2. A few examples of micrographs of yeast strains before and during fluorescence emission. Increasing intracellular lipid yield evaluated by Nile Red before (A, C, E and G) and during fluorescence emission (B, D, F and H) photographed with a Wild MP 552 camera (Leica). A and B = Debaryomyces hansenii DBVPG 3326; C and D = Zygosaccharomyces bisporus DBVPG 3018; E and F = Magnusiomyces capitatus DBVPG 3250; G and H = Naganishia albida DBVPG 4919

  • MOESM4 of Study of Holtermanniella wattica, Leucosporidium creatinivorum, Naganishia adeliensis, Solicoccozyma aeria, and Solicoccozyma terricola for their lipogenic aptitude from different carbon sources
    2016
    Co-Authors: Sara Filippucci, Giorgia Tasselli, Benedetta Turchetti, Andrea Onofri, Alessandro Scardua, Simone Mauro, Maria Cramarossa, Davide Perini, Luca Forti, Pietro Buzzini
    Abstract:

    Additional file 4: Figure S3. High resolution images of NR staining of Leucosporidium creatinivorum DBVPG 4794 (incubated at 20 °C), Naganishia adeliensis DBVPG 5195 and Solicoccozyma terricola DBVPG 5870 (both at 25 °C). High resolution images of Nile Red staining of Naganishia adeliensis DBVPG 5195 (A and B, incubated at 25 °C), Solicoccozyma terricola DBVPG 5870 (C and D, 25 °C) and Leucosporidium creatinivorum DBVPG 4794 (E and F, 20 °C). Photographs were captured before (A, C and E) and during fluorescence emission (B, D and F) with an UV epifluorescence microscope Olympus BX53 (Olympus Co., Centre Valley, PA, USA) equipped with excitation filter BP470-500 and barrier filter BA515-560 and a XC50 camera (Olympus Co.)

Giorgia Tasselli - One of the best experts on this subject based on the ideXlab platform.

  • yeast lipids from cardoon stalks stranded driftwood and olive tree pruning residues as possible extra sources of oils for producing biofuels and biochemicals
    Biotechnology for Biofuels, 2018
    Co-Authors: Giorgia Tasselli, Sara Filippucci, E Borsella, Silvia Dantonio, Mattia Gelosia, Gianluca Cavalaglio, Benedetta Turchetti, Ciro Sannino, Andrea Onofri, Silvio Mastrolitti
    Abstract:

    Some lignocellulosic biomass feedstocks occur in Mediterranean Countries. They are still largely unexploited and cause considerable problems due to the lack of cost-effective harvesting, storage and disposal technologies. Recent studies found that some basidiomycetous yeasts are able to accumulate high amount of intracellular lipids for biorefinery processes (i.e., biofuels and biochemicals). Accordingly, the above biomass feedstocks could be used as carbon sources (after their pre-treatment and hydrolysis) for lipid accumulation by oleaginous yeasts. Cardoon stalks, stranded driftwood and olive tree pruning residues were pre-treated with steam-explosion and enzymatic hydrolysis for releasing free mono- and oligosaccharides. Lipid accumulation tests were performed at two temperatures (20 and 25 °C) using Leucosporidium creatinivorum DBVPG 4794, Naganishia adeliensis DBVPG 5195 and Solicoccozyma terricola DBVPG 5870. S. terricola grown on cardoon stalks at 20 °C exhibited the highest lipid production (13.20 g/l), a lipid yield (28.95%) close to the maximum theoretical value and a lipid composition similar to that found in palm oil. On the contrary, N. adeliensis grown on stranded driftwood and olive tree pruning residues exhibited a lipid composition similar to those of olive and almonds oils. A predictive evaluation of the physical properties of the potential biodiesel obtainable by lipids produced by tested yeast strains has been reported and discussed. Lipids produced by some basidiomycetous yeasts grown on Mediterranean lignocellulosic biomass feedstocks could be used as supplementary sources of oils for producing biofuels and biochemicals.

  • Yeast lipids from cardoon stalks, stranded driftwood and olive tree pruning residues as possible extra sources of oils for producing biofuels and biochemicals
    BMC, 2018
    Co-Authors: Giorgia Tasselli, Sara Filippucci, E Borsella, Mattia Gelosia, Gianluca Cavalaglio, Benedetta Turchetti, Ciro Sannino, Andrea Onofri, Silvia D’antonio, Silvio Mastrolitti
    Abstract:

    Abstract Background Some lignocellulosic biomass feedstocks occur in Mediterranean Countries. They are still largely unexploited and cause considerable problems due to the lack of cost-effective harvesting, storage and disposal technologies. Recent studies found that some basidiomycetous yeasts are able to accumulate high amount of intracellular lipids for biorefinery processes (i.e., biofuels and biochemicals). Accordingly, the above biomass feedstocks could be used as carbon sources (after their pre-treatment and hydrolysis) for lipid accumulation by oleaginous yeasts. Results Cardoon stalks, stranded driftwood and olive tree pruning residues were pre-treated with steam-explosion and enzymatic hydrolysis for releasing free mono- and oligosaccharides. Lipid accumulation tests were performed at two temperatures (20 and 25 °C) using Leucosporidium creatinivorum DBVPG 4794, Naganishia adeliensis DBVPG 5195 and Solicoccozyma terricola DBVPG 5870. S. terricola grown on cardoon stalks at 20 °C exhibited the highest lipid production (13.20 g/l), a lipid yield (28.95%) close to the maximum theoretical value and a lipid composition similar to that found in palm oil. On the contrary, N. adeliensis grown on stranded driftwood and olive tree pruning residues exhibited a lipid composition similar to those of olive and almonds oils. A predictive evaluation of the physical properties of the potential biodiesel obtainable by lipids produced by tested yeast strains has been reported and discussed. Conclusions Lipids produced by some basidiomycetous yeasts grown on Mediterranean lignocellulosic biomass feedstocks could be used as supplementary sources of oils for producing biofuels and biochemicals

  • study of holtermanniella wattica Leucosporidium creatinivorum naganishia adeliensis solicoccozyma aeria and solicoccozyma terricola for their lipogenic aptitude from different carbon sources
    Biotechnology for Biofuels, 2016
    Co-Authors: Sara Filippucci, Giorgia Tasselli, Benedetta Turchetti, Andrea Onofri, Alessandro Scardua, Simone Mauro, Maria Cramarossa, Davide Perini, Luca Forti, Pietro Buzzini
    Abstract:

    The ability of some microorganisms to accumulate lipids is well known; however, only recently the number of studies on microbial lipid biosynthesis for obtaining oleochemical products, namely biofuels and some building blocks for chemistry, is rapidly and spectacularly increased. Since 1990s, some oleaginous yeasts were studied for their ability to accumulate lipids up to 60–70% of their dry weight. Due to the vast array of engineering techniques currently available, the recombinant DNA technology was the main approach followed so far for obtaining lipid-overproducing yeasts, mainly belonging to the Yarrowia lipolytica. However, an alternative approach can be offered by worldwide diversity as source of novel oleaginous yeasts. Lipogenic aptitude of a number of yeast strains has been reviewed, but many of these studies utilized a limited number of species and/or different culture conditions that make impossible the comparison of different results. Accordingly, the lipogenic aptitude inside the yeast world is still far from being fully explored, and finding new oleaginous yeast species can acquire a strategic importance. Holtermanniella wattica, Leucosporidium creatinivorum, Naganishia adeliensis, Solicoccozyma aeria, and Solicoccozyma terricola strains were selected as a result of a large-scale screening on 706 yeasts (both Ascomycota and Basidiomycota). Lipid yields and fatty acid profiles of selected strains were evaluated at 20 and 25 °C on glucose, and on glycerol, xylose, galactose, sucrose, maltose, and cellobiose. A variable fatty acid profile was observed in dependence of both temperature and different carbon sources. On the whole, L. creatinivorum exhibited the highest performances: total lipid yield (YL) >7 g/l on glucose and glycerol, % of intracellular lipids on cell biomass (YL/DW) >70% at 20 °C on glucose, lipid coefficient (YL/Glu) around 20% on glucose, and daily productivity (YL/d) on glucose and sucrose >1.6 g/(l*d). This study provides some meaningful information about the lipogenic ability of some yeast species. Variable lipid yields and fatty acid profiles were observed in dependence of both temperature and different carbon sources. L. creatinivorum exhibited the highest lipogenic performances.

  • MOESM3 of Study of Holtermanniella wattica, Leucosporidium creatinivorum, Naganishia adeliensis, Solicoccozyma aeria, and Solicoccozyma terricola for their lipogenic aptitude from different carbon sources
    2016
    Co-Authors: Sara Filippucci, Giorgia Tasselli, Benedetta Turchetti, Andrea Onofri, Alessandro Scardua, Simone Mauro, Maria Cramarossa, Davide Perini, Luca Forti, Pietro Buzzini
    Abstract:

    Additional file 3: Figure S2. A few examples of micrographs of yeast strains before and during fluorescence emission. Increasing intracellular lipid yield evaluated by Nile Red before (A, C, E and G) and during fluorescence emission (B, D, F and H) photographed with a Wild MP 552 camera (Leica). A and B = Debaryomyces hansenii DBVPG 3326; C and D = Zygosaccharomyces bisporus DBVPG 3018; E and F = Magnusiomyces capitatus DBVPG 3250; G and H = Naganishia albida DBVPG 4919

  • MOESM4 of Study of Holtermanniella wattica, Leucosporidium creatinivorum, Naganishia adeliensis, Solicoccozyma aeria, and Solicoccozyma terricola for their lipogenic aptitude from different carbon sources
    2016
    Co-Authors: Sara Filippucci, Giorgia Tasselli, Benedetta Turchetti, Andrea Onofri, Alessandro Scardua, Simone Mauro, Maria Cramarossa, Davide Perini, Luca Forti, Pietro Buzzini
    Abstract:

    Additional file 4: Figure S3. High resolution images of NR staining of Leucosporidium creatinivorum DBVPG 4794 (incubated at 20 °C), Naganishia adeliensis DBVPG 5195 and Solicoccozyma terricola DBVPG 5870 (both at 25 °C). High resolution images of Nile Red staining of Naganishia adeliensis DBVPG 5195 (A and B, incubated at 25 °C), Solicoccozyma terricola DBVPG 5870 (C and D, 25 °C) and Leucosporidium creatinivorum DBVPG 4794 (E and F, 20 °C). Photographs were captured before (A, C and E) and during fluorescence emission (B, D and F) with an UV epifluorescence microscope Olympus BX53 (Olympus Co., Centre Valley, PA, USA) equipped with excitation filter BP470-500 and barrier filter BA515-560 and a XC50 camera (Olympus Co.)