The Experts below are selected from a list of 174 Experts worldwide ranked by ideXlab platform
Graciette Matioli - One of the best experts on this subject based on the ideXlab platform.
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immobilization of commercial Cyclomaltodextrin Glucanotransferase into controlled pore silica by the anchorage method and covalent bonding
Process Biochemistry, 2019Co-Authors: Gabriela Gregolin Gimenez, Richard Marllon Silva, Carolina Pereira Francisco, Fabiana Dos Santos Rando, Joao Henrique Dantas, Hâmara Milaneze De Souza, Graciette MatioliAbstract:Abstract Cyclomaltodextrin Glucanotransferase (CGTase) is used mainly for the industrial production of cyclodextrins (CDs). Their potential use and availability have played a decisive role in increasing demand for CDs. The aim of the present study was to immobilize the commercial CGTase (Toruzyme® 3.0L) into controlled pore silica by surface anchoring (SCGT-A) and covalent bonding (SCGT-CB) and to apply the immobilized enzymes in the production of CDs. The SCGT-A method proved to be efficient due to its short immobilization time and, it was possible to immobilize 89.63% of CGTase, in terms of activity. Commercial CGTase immobilized by SCGT-CB provided an immobilization yield of 48.44%, also in terms of activity. There was an increase in total CD production of 60.97% and 34.90% for SCGT-A and SCGT-CB, respectively, in a reaction medium with 10% ethanol, compared to the same media without ethanol. Higher CD production occured at 24 and 48 h for SCGT-A and SCGT-CB methods, respectively. The use of SCGT-A was more efficient than SCGT-CB and free enzyme (CGT-F) after 24 h of reaction. The increase production of CDs obtained by the SCGT-A method and the demonstrated continuous production of CDs with the SCGT-CB method make the use of immobilized CGTase viable for industrial applications.
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sequencing cloning and heterologous expression of Cyclomaltodextrin Glucanotransferase of bacillus firmus strain 37 in bacillus subtilis wb800
Bioprocess and Biosystems Engineering, 2019Co-Authors: Gabriela Gregolin Gimenez, Susana Alicia Ferrarotti, Hernan Costa, Quirino Alves De Lima Neto, Maria Aparecida Fernandez, Graciette MatioliAbstract:Bacillusfirmus strain 37 produces the Cyclomaltodextrin Glucanotransferase (CGTase) enzyme and CGTase produces cyclodextrins (CDs) through a starch cyclization reaction. The strategy for the cloning and expression of recombinant CGTase is a potentially viable alternative for the economically viable production of CGTase for use in industrial processes. The present study used Bacillus subtilis WB800 as a bacterial expression host for the production of recombinant CGTase cloned from the CGTase gene of B. firmus strain 37. The CGTase gene was cloned in TOPO-TA® plasmid, which was transformed in Escherichia coli DH5α. The subcloning was carried out with pWB980 plasmid and transformation in B. subtilis WB800. The 2xYT medium was the most suitable for the production of recombinant CGTase. The enzymatic activity of the crude extract of the recombinant CGTase of B. subtilis WB800 was 1.33 µmol β-CD/min/mL, or 7.4 times greater than the enzymatic activity of the crude extract of CGTase obtained from the wild strain. Following purification, the recombinant CGTase exhibited an enzymatic activity of 157.78 µmol β-CD/min/mL, while the activity of the CGTase from the wild strain was 9.54 µmol β-CD/min/mL. When optimal CDs production conditions for the CGTase from B. firmus strain 37 were used, it was observed that the catalytic properties of the CGTase enzymes were equivalent. The strategy for the cloning and expression of CGTase in B. subtilis WB800 was efficient, with the production of greater quantities of CGTase than with the wild strain, offering essential data for the large-scale production of the recombinant enzyme.
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bovine bone charcoal as support material for immobilization of bacillus firmus strain 37 and production of Cyclomaltodextrin Glucanotransferase by batch fermentation in a fluidized bed
Advances in Chemical Engineering and Science, 2018Co-Authors: Larissa Albunio Silva, Gisella Maria Zanin, Graciette Matioli, Bruno Cesar Bieli, Osvaldo Valarini, Flavio Faria De MoraesAbstract:The process described in the present work uses air supplementation in a fluidized bed reactor containing Bacillus firmus strain 37 immobilized on active bovine bone charcoal, to produce by batch fermentation the enzyme CGTase (Cyclomaltodextrin-Glucanotransferase). Three different aeration rates were evaluated. The maximum CGTase activity was achieved after 120 hours of fermentation with aeration rate of 2 vvm and was equal to 2.48 U/mL. When 0.5 and 1 vvm were used the enzymatic activities achieved 1.1 and 0.57 U/mL, respectively. Bovine bone charcoal was characterized in terms of surface area, pore size and volume. To the best of our knowledge, the immobilization of microorganism cells in bovine bone charcoal for CGTase production has not been reported in the literature. Our results showed that fluidized bed reactor allows retaining high concentration of biomass, improving biomass-substrate contact and operation at low residence times, which resulted in improved enzyme production. Therefore, the process as proposed has great potential for industrial development.
Tadayuki Imanaka - One of the best experts on this subject based on the ideXlab platform.
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purification and characterization of an extremely thermostable Cyclomaltodextrin Glucanotransferase from a newly isolated hyperthermophilic archaeon a thermococcus sp
Applied and Environmental Microbiology, 1999Co-Authors: Yoshihisa Tachibana, Akiko Kuramura, Naoki Shirasaka, Yuji Suzuki, Tomoko Yamamoto, Shinsuke Fujiwara, Masahiro Takagi, Tadayuki ImanakaAbstract:The extremely thermophilic anaerobic archaeon strain B1001 was isolated from a hot-spring environment in Japan. The cells were irregular cocci, 0.5 to 1.0 μm in diameter. The new isolate grew at temperatures between 60 and 95°C (optimum, 85°C), from pH 5.0 to 9.0 (optimum, pH 7.0), and from 1.0 to 6.0% NaCl (optimum, 2.0%). The G+C content of the genomic DNA was 43.0 mol%. The 16S rRNA gene sequencing of strain B1001 indicated that it belongs to the genus Thermococcus. During growth on starch, the strain produced a thermostable Cyclomaltodextrin Glucanotransferase (CGTase). The enzyme was purified 1,750-fold, and the molecular mass was determined to be 83 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Incubation at 120°C with SDS and 2-mercaptoethanol was required for complete unfolding. The optimum temperatures for starch-degrading activity and cyclodextrin synthesis activity were 110 and 90 to 100°C, respectively. The optimum pH for enzyme activity was pH 5.0 to 5.5. At pH 5.0, the half-life of the enzyme was 40 min at 110°C. The enzyme formed mainly α-cyclodextrin with small amounts of β- and γ-cyclodextrins from starch. This is the first report on the presence of the extremely thermostable CGTase from hyperthermophilic archaea.
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purification and characterization of an extremely thermostable Cyclomaltodextrin Glucanotransferase from a newly isolated hyperthermophilic archaeon a thermococcus sp
Applied and Environmental Microbiology, 1999Co-Authors: Yoshihisa Tachibana, Akiko Kuramura, Naoki Shirasaka, Yuji Suzuki, Tomoko Yamamoto, Shinsuke Fujiwara, Masahiro Takagi, Tadayuki ImanakaAbstract:The extremely thermophilic anaerobic archaeon strain B1001 was isolated from a hot-spring environment in Japan. The cells were irregular cocci, 0.5 to 1.0 micrometers in diameter. The new isolate grew at temperatures between 60 and 95 degrees C (optimum, 85 degrees C), from pH 5.0 to 9.0 (optimum, pH 7.0), and from 1.0 to 6.0% NaCl (optimum, 2.0%). The G+C content of the genomic DNA was 43.0 mol%. The 16S rRNA gene sequencing of strain B1001 indicated that it belongs to the genus Thermococcus. During growth on starch, the strain produced a thermostable Cyclomaltodextrin Glucanotransferase (CGTase). The enzyme was purified 1,750-fold, and the molecular mass was determined to be 83 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Incubation at 120 degrees C with SDS and 2-mercaptoethanol was required for complete unfolding. The optimum temperatures for starch-degrading activity and cyclodextrin synthesis activity were 110 and 90 to 100 degrees C, respectively. The optimum pH for enzyme activity was pH 5.0 to 5.5. At pH 5.0, the half-life of the enzyme was 40 min at 110 degrees C. The enzyme formed mainly alpha-cyclodextrin with small amounts of beta- and gamma-cyclodextrins from starch. This is the first report on the presence of the extremely thermostable CGTase from hyperthermophilic archaea.
Gabriela Gregolin Gimenez - One of the best experts on this subject based on the ideXlab platform.
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immobilization of commercial Cyclomaltodextrin Glucanotransferase into controlled pore silica by the anchorage method and covalent bonding
Process Biochemistry, 2019Co-Authors: Gabriela Gregolin Gimenez, Richard Marllon Silva, Carolina Pereira Francisco, Fabiana Dos Santos Rando, Joao Henrique Dantas, Hâmara Milaneze De Souza, Graciette MatioliAbstract:Abstract Cyclomaltodextrin Glucanotransferase (CGTase) is used mainly for the industrial production of cyclodextrins (CDs). Their potential use and availability have played a decisive role in increasing demand for CDs. The aim of the present study was to immobilize the commercial CGTase (Toruzyme® 3.0L) into controlled pore silica by surface anchoring (SCGT-A) and covalent bonding (SCGT-CB) and to apply the immobilized enzymes in the production of CDs. The SCGT-A method proved to be efficient due to its short immobilization time and, it was possible to immobilize 89.63% of CGTase, in terms of activity. Commercial CGTase immobilized by SCGT-CB provided an immobilization yield of 48.44%, also in terms of activity. There was an increase in total CD production of 60.97% and 34.90% for SCGT-A and SCGT-CB, respectively, in a reaction medium with 10% ethanol, compared to the same media without ethanol. Higher CD production occured at 24 and 48 h for SCGT-A and SCGT-CB methods, respectively. The use of SCGT-A was more efficient than SCGT-CB and free enzyme (CGT-F) after 24 h of reaction. The increase production of CDs obtained by the SCGT-A method and the demonstrated continuous production of CDs with the SCGT-CB method make the use of immobilized CGTase viable for industrial applications.
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sequencing cloning and heterologous expression of Cyclomaltodextrin Glucanotransferase of bacillus firmus strain 37 in bacillus subtilis wb800
Bioprocess and Biosystems Engineering, 2019Co-Authors: Gabriela Gregolin Gimenez, Susana Alicia Ferrarotti, Hernan Costa, Quirino Alves De Lima Neto, Maria Aparecida Fernandez, Graciette MatioliAbstract:Bacillusfirmus strain 37 produces the Cyclomaltodextrin Glucanotransferase (CGTase) enzyme and CGTase produces cyclodextrins (CDs) through a starch cyclization reaction. The strategy for the cloning and expression of recombinant CGTase is a potentially viable alternative for the economically viable production of CGTase for use in industrial processes. The present study used Bacillus subtilis WB800 as a bacterial expression host for the production of recombinant CGTase cloned from the CGTase gene of B. firmus strain 37. The CGTase gene was cloned in TOPO-TA® plasmid, which was transformed in Escherichia coli DH5α. The subcloning was carried out with pWB980 plasmid and transformation in B. subtilis WB800. The 2xYT medium was the most suitable for the production of recombinant CGTase. The enzymatic activity of the crude extract of the recombinant CGTase of B. subtilis WB800 was 1.33 µmol β-CD/min/mL, or 7.4 times greater than the enzymatic activity of the crude extract of CGTase obtained from the wild strain. Following purification, the recombinant CGTase exhibited an enzymatic activity of 157.78 µmol β-CD/min/mL, while the activity of the CGTase from the wild strain was 9.54 µmol β-CD/min/mL. When optimal CDs production conditions for the CGTase from B. firmus strain 37 were used, it was observed that the catalytic properties of the CGTase enzymes were equivalent. The strategy for the cloning and expression of CGTase in B. subtilis WB800 was efficient, with the production of greater quantities of CGTase than with the wild strain, offering essential data for the large-scale production of the recombinant enzyme.
Yoshihisa Tachibana - One of the best experts on this subject based on the ideXlab platform.
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purification and characterization of an extremely thermostable Cyclomaltodextrin Glucanotransferase from a newly isolated hyperthermophilic archaeon a thermococcus sp
Applied and Environmental Microbiology, 1999Co-Authors: Yoshihisa Tachibana, Akiko Kuramura, Naoki Shirasaka, Yuji Suzuki, Tomoko Yamamoto, Shinsuke Fujiwara, Masahiro Takagi, Tadayuki ImanakaAbstract:The extremely thermophilic anaerobic archaeon strain B1001 was isolated from a hot-spring environment in Japan. The cells were irregular cocci, 0.5 to 1.0 μm in diameter. The new isolate grew at temperatures between 60 and 95°C (optimum, 85°C), from pH 5.0 to 9.0 (optimum, pH 7.0), and from 1.0 to 6.0% NaCl (optimum, 2.0%). The G+C content of the genomic DNA was 43.0 mol%. The 16S rRNA gene sequencing of strain B1001 indicated that it belongs to the genus Thermococcus. During growth on starch, the strain produced a thermostable Cyclomaltodextrin Glucanotransferase (CGTase). The enzyme was purified 1,750-fold, and the molecular mass was determined to be 83 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Incubation at 120°C with SDS and 2-mercaptoethanol was required for complete unfolding. The optimum temperatures for starch-degrading activity and cyclodextrin synthesis activity were 110 and 90 to 100°C, respectively. The optimum pH for enzyme activity was pH 5.0 to 5.5. At pH 5.0, the half-life of the enzyme was 40 min at 110°C. The enzyme formed mainly α-cyclodextrin with small amounts of β- and γ-cyclodextrins from starch. This is the first report on the presence of the extremely thermostable CGTase from hyperthermophilic archaea.
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purification and characterization of an extremely thermostable Cyclomaltodextrin Glucanotransferase from a newly isolated hyperthermophilic archaeon a thermococcus sp
Applied and Environmental Microbiology, 1999Co-Authors: Yoshihisa Tachibana, Akiko Kuramura, Naoki Shirasaka, Yuji Suzuki, Tomoko Yamamoto, Shinsuke Fujiwara, Masahiro Takagi, Tadayuki ImanakaAbstract:The extremely thermophilic anaerobic archaeon strain B1001 was isolated from a hot-spring environment in Japan. The cells were irregular cocci, 0.5 to 1.0 micrometers in diameter. The new isolate grew at temperatures between 60 and 95 degrees C (optimum, 85 degrees C), from pH 5.0 to 9.0 (optimum, pH 7.0), and from 1.0 to 6.0% NaCl (optimum, 2.0%). The G+C content of the genomic DNA was 43.0 mol%. The 16S rRNA gene sequencing of strain B1001 indicated that it belongs to the genus Thermococcus. During growth on starch, the strain produced a thermostable Cyclomaltodextrin Glucanotransferase (CGTase). The enzyme was purified 1,750-fold, and the molecular mass was determined to be 83 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Incubation at 120 degrees C with SDS and 2-mercaptoethanol was required for complete unfolding. The optimum temperatures for starch-degrading activity and cyclodextrin synthesis activity were 110 and 90 to 100 degrees C, respectively. The optimum pH for enzyme activity was pH 5.0 to 5.5. At pH 5.0, the half-life of the enzyme was 40 min at 110 degrees C. The enzyme formed mainly alpha-cyclodextrin with small amounts of beta- and gamma-cyclodextrins from starch. This is the first report on the presence of the extremely thermostable CGTase from hyperthermophilic archaea.
Flavio Faria De Moraes - One of the best experts on this subject based on the ideXlab platform.
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bovine bone charcoal as support material for immobilization of bacillus firmus strain 37 and production of Cyclomaltodextrin Glucanotransferase by batch fermentation in a fluidized bed
Advances in Chemical Engineering and Science, 2018Co-Authors: Larissa Albunio Silva, Gisella Maria Zanin, Graciette Matioli, Bruno Cesar Bieli, Osvaldo Valarini, Flavio Faria De MoraesAbstract:The process described in the present work uses air supplementation in a fluidized bed reactor containing Bacillus firmus strain 37 immobilized on active bovine bone charcoal, to produce by batch fermentation the enzyme CGTase (Cyclomaltodextrin-Glucanotransferase). Three different aeration rates were evaluated. The maximum CGTase activity was achieved after 120 hours of fermentation with aeration rate of 2 vvm and was equal to 2.48 U/mL. When 0.5 and 1 vvm were used the enzymatic activities achieved 1.1 and 0.57 U/mL, respectively. Bovine bone charcoal was characterized in terms of surface area, pore size and volume. To the best of our knowledge, the immobilization of microorganism cells in bovine bone charcoal for CGTase production has not been reported in the literature. Our results showed that fluidized bed reactor allows retaining high concentration of biomass, improving biomass-substrate contact and operation at low residence times, which resulted in improved enzyme production. Therefore, the process as proposed has great potential for industrial development.
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Methods and Supports for Immobilization and Stabilization of Cyclomaltodextrin Glucanotransferase from Thermoanaerobacter
Applied Biochemistry and Biotechnology, 2008Co-Authors: Ana Elisa Amud, Paulo Waldir Tardioli, Flavio Faria De Moraes, Gercio Rodrigo Presa Da Silva, Cleide Mara Faria Soares, Gisella Maria ZaninAbstract:Thermoanaerobacter Cyclomaltodextrin Glucanotransferase (CGTase) was immobilized using different supports and immobilization methods to study the effect on activity recovery. The enzyme covalently attached into glyoxyl-silica showed low activity recovery of 1.5%. The hydrophobic adsorption of the enzyme on Octadecyl-Sepabeads yielded also low activity recovery, 3.83%, and the enzyme could easily leak from the support at low ionic strength, although the immobilization yield was satisfactory, approximately 76%. The CGTase encapsulated in a sol–gel matrix gave an activity recovery of 6.94% and maximum cyclization activity at 60 °C, at pH 6.0. The half-time life at 60 °C, pH 6.0, in the presence of substrate was 100 min, which was lower than that of the free enzyme. The best activity recovery in this work (6.94%) is approximately five times smaller than that obtained previously using glyoxyl-agarose as support and covalent immobilization. Thus, the best support and method we tested so far for immobilization of CGTase is covalent attachment on glyoxyl-agarose.
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characterization of thermoanaerobacter Cyclomaltodextrin Glucanotransferase immobilized on glyoxyl agarose
Enzyme and Microbial Technology, 2006Co-Authors: Paulo Waldir Tardioli, Gisella Maria Zanin, Flavio Faria De MoraesAbstract:Abstract This paper presents the immobilization of the Thermoanaerobacter Cyclomaltodextrin Glucanotransferase (CGTase) enzyme into cross-linked 6% agarose beads activated by high density of linear aldehyde groups (glyoxyl-agarose) that allow the establishment of multi-attachment enzyme-support bonds. The immobilization conditions were 25 °C, pH 10 and 5 h of contact time. The immobilization yield was almost 100% and the activity recovery was ca. 32%. The biocatalyst at 85 °C was capable of producing cyclodextrins (CDs) from dextrin or soluble starch (both at 1% (w/v)) at a greater rate than the soluble enzyme. In addition, the biocatalyst maintained 90% of its initial activity after 5 h at 85 °C. The maximum conversion of dextrin to β-CD and γ-CD (total mass of produced CDs/substrate initial mass × 100) was 29% both for the soluble and immobilized enzymes. Using starch as substrate the maximum starch conversion to β-CD and γ-CD was 29% and 38%, for the immobilized and soluble enzyme, respectively. The β-CD selectivity yield [mass of β-CD produced/(mass of β-CD produced + mass of γ-CD produced) × 100] increased from 67.9% for the free enzyme to 85.4% for the immobilized CGTase.
