High level production of tyrosinase in recombinant Escherichia coli
Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particu...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Ren, Qun [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2013 |
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| Schlagwörter: |
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| Anmerkung: |
© Ren et al.; licensee BioMed Central Ltd. 2013 |
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| Übergeordnetes Werk: |
Enthalten in: BMC biotechnology - London : BioMed Central, 2001, 13(2013), 1 vom: 27. Feb. |
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| Übergeordnetes Werk: |
volume:13 ; year:2013 ; number:1 ; day:27 ; month:02 |
| Links: |
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| DOI / URN: |
10.1186/1472-6750-13-18 |
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| Katalog-ID: |
SPR028407180 |
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| 245 | 1 | 0 | |a High level production of tyrosinase in recombinant Escherichia coli |
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| 520 | |a Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. | ||
| 650 | 4 | |a Tyrosinase |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Recombinant protein production |7 (dpeaa)DE-He213 | |
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| 650 | 4 | |a Fed batch culture |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Bioprocess engineering |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Exponential feeding |7 (dpeaa)DE-He213 | |
| 700 | 1 | |a Henes, Bernhard |4 aut | |
| 700 | 1 | |a Fairhead, Michael |4 aut | |
| 700 | 1 | |a Thöny-Meyer, Linda |4 aut | |
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10.1186/1472-6750-13-18 doi (DE-627)SPR028407180 (SPR)1472-6750-13-18-e DE-627 ger DE-627 rakwb eng Ren, Qun verfasserin aut High level production of tyrosinase in recombinant Escherichia coli 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Ren et al.; licensee BioMed Central Ltd. 2013 Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. Tyrosinase (dpeaa)DE-He213 Recombinant protein production (dpeaa)DE-He213 High cell density (dpeaa)DE-He213 Fed batch culture (dpeaa)DE-He213 Bioprocess engineering (dpeaa)DE-He213 Exponential feeding (dpeaa)DE-He213 Henes, Bernhard aut Fairhead, Michael aut Thöny-Meyer, Linda aut Enthalten in BMC biotechnology London : BioMed Central, 2001 13(2013), 1 vom: 27. Feb. (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:13 year:2013 number:1 day:27 month:02 https://dx.doi.org/10.1186/1472-6750-13-18 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2013 1 27 02 |
| spelling |
10.1186/1472-6750-13-18 doi (DE-627)SPR028407180 (SPR)1472-6750-13-18-e DE-627 ger DE-627 rakwb eng Ren, Qun verfasserin aut High level production of tyrosinase in recombinant Escherichia coli 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Ren et al.; licensee BioMed Central Ltd. 2013 Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. Tyrosinase (dpeaa)DE-He213 Recombinant protein production (dpeaa)DE-He213 High cell density (dpeaa)DE-He213 Fed batch culture (dpeaa)DE-He213 Bioprocess engineering (dpeaa)DE-He213 Exponential feeding (dpeaa)DE-He213 Henes, Bernhard aut Fairhead, Michael aut Thöny-Meyer, Linda aut Enthalten in BMC biotechnology London : BioMed Central, 2001 13(2013), 1 vom: 27. Feb. (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:13 year:2013 number:1 day:27 month:02 https://dx.doi.org/10.1186/1472-6750-13-18 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2013 1 27 02 |
| allfields_unstemmed |
10.1186/1472-6750-13-18 doi (DE-627)SPR028407180 (SPR)1472-6750-13-18-e DE-627 ger DE-627 rakwb eng Ren, Qun verfasserin aut High level production of tyrosinase in recombinant Escherichia coli 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Ren et al.; licensee BioMed Central Ltd. 2013 Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. Tyrosinase (dpeaa)DE-He213 Recombinant protein production (dpeaa)DE-He213 High cell density (dpeaa)DE-He213 Fed batch culture (dpeaa)DE-He213 Bioprocess engineering (dpeaa)DE-He213 Exponential feeding (dpeaa)DE-He213 Henes, Bernhard aut Fairhead, Michael aut Thöny-Meyer, Linda aut Enthalten in BMC biotechnology London : BioMed Central, 2001 13(2013), 1 vom: 27. Feb. (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:13 year:2013 number:1 day:27 month:02 https://dx.doi.org/10.1186/1472-6750-13-18 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2013 1 27 02 |
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10.1186/1472-6750-13-18 doi (DE-627)SPR028407180 (SPR)1472-6750-13-18-e DE-627 ger DE-627 rakwb eng Ren, Qun verfasserin aut High level production of tyrosinase in recombinant Escherichia coli 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Ren et al.; licensee BioMed Central Ltd. 2013 Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. Tyrosinase (dpeaa)DE-He213 Recombinant protein production (dpeaa)DE-He213 High cell density (dpeaa)DE-He213 Fed batch culture (dpeaa)DE-He213 Bioprocess engineering (dpeaa)DE-He213 Exponential feeding (dpeaa)DE-He213 Henes, Bernhard aut Fairhead, Michael aut Thöny-Meyer, Linda aut Enthalten in BMC biotechnology London : BioMed Central, 2001 13(2013), 1 vom: 27. Feb. (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:13 year:2013 number:1 day:27 month:02 https://dx.doi.org/10.1186/1472-6750-13-18 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2013 1 27 02 |
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10.1186/1472-6750-13-18 doi (DE-627)SPR028407180 (SPR)1472-6750-13-18-e DE-627 ger DE-627 rakwb eng Ren, Qun verfasserin aut High level production of tyrosinase in recombinant Escherichia coli 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Ren et al.; licensee BioMed Central Ltd. 2013 Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. Tyrosinase (dpeaa)DE-He213 Recombinant protein production (dpeaa)DE-He213 High cell density (dpeaa)DE-He213 Fed batch culture (dpeaa)DE-He213 Bioprocess engineering (dpeaa)DE-He213 Exponential feeding (dpeaa)DE-He213 Henes, Bernhard aut Fairhead, Michael aut Thöny-Meyer, Linda aut Enthalten in BMC biotechnology London : BioMed Central, 2001 13(2013), 1 vom: 27. Feb. (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:13 year:2013 number:1 day:27 month:02 https://dx.doi.org/10.1186/1472-6750-13-18 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2013 1 27 02 |
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Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. 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High level production of tyrosinase in recombinant Escherichia coli Tyrosinase (dpeaa)DE-He213 Recombinant protein production (dpeaa)DE-He213 High cell density (dpeaa)DE-He213 Fed batch culture (dpeaa)DE-He213 Bioprocess engineering (dpeaa)DE-He213 Exponential feeding (dpeaa)DE-He213 |
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high level production of tyrosinase in recombinant escherichia coli |
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High level production of tyrosinase in recombinant Escherichia coli |
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Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. © Ren et al.; licensee BioMed Central Ltd. 2013 |
| abstractGer |
Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. © Ren et al.; licensee BioMed Central Ltd. 2013 |
| abstract_unstemmed |
Background Tyrosinase is a bifunctional enzyme that catalyzes both the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the subsequent oxidation of the diphenols to o-quinones (diphenolase activity). Due to the potential applications of tyrosinase in biotechnology, in particular in biocatalysis and for biosensors, it is desirable to develop a suitable low-cost process for efficient production of this enzyme. So far, the best production yield reported for tyrosinase was about 1 g $ L^{-1} $, which was achieved by cultivating the filamentous fungus Trichoderma reesei for 6 days. Results In this work, tyrosinase from Verrucomicrobium spinosum was expressed in Escherichia coli and its production was studied in both batch and fed-batch cultivations. Effects of various key cultivation parameters on tyrosinase production were first examined in batch cultures to identify optimal conditions. It was found that a culture temperature of 32 °C and induction at the late growth stage were favorable, leading to a highest tyrosinase activity of 0.76 U $ mL^{-1} $. The fed-batch process was performed by using an exponential feeding strategy to achieve high cell density. With the fed-batch process, a final biomass concentration of 37 g $ L^{-1} $ (based on optical density) and a tyrosinase activity of 13 U $ mL^{-1} $ were obtained in 28 hours, leading to a yield of active tyrosinase of about 3 g $ L^{-1} $. The highest overall volumetric productivity of 103 mg of active tyrosinase per liter and hour (corresponding to 464 mU $ L^{-1} $ $ h^{-1} $) was determined, which is approximately 15 times higher than that obtained in batch cultures. Conclusions We have successfully expressed and produced gram quantities per liter of active tyrosinase in recombinant E. coli by optimizing the expression conditions and fed-batch cultivation strategy. Exponential feed of substrate helped to prolong the exponential phase of growth, to reduce the fermentation time and thus the cost. A specific tyrosinase production rate of 103 mg $ L^{−1} $ $ h^{−1} $ and a maximum volumetric activity of 464 mU $ L^{−1} $ $ h^{-1} $ were achieved in this study. These levels have not been reported previously. © Ren et al.; licensee BioMed Central Ltd. 2013 |
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| score |
7.4010954 |