Media improvement for 10 L bioreactor production of rPOXA 1B laccase by P. pastoris
| العنوان: | Media improvement for 10 L bioreactor production of rPOXA 1B laccase by P. pastoris |
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| المؤلفون: | Ángela M. Cardozo-Bernal, Dennis J. Díaz-Rincón, Carlos J. Alméciga-Díaz, Edwin D. Morales-Álvarez, Claudia L. Cuervo-Patiño, Leidy D. Ardila-Leal, Claudia M. Rivera-Hoyos, Aura M. Pedroza-Rodríguez, Raúl A. Poutou-Piñales, Balkys Quevedo-Hidalgo, Alexander Rodríguez-López, Diego A. Albarracín-Pardo |
| المصدر: | 3 Biotech. 9 |
| بيانات النشر: | Springer Science and Business Media LLC, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | chemistry.chemical_classification, Laccase, ABTS, Chromatography, biology, Chemistry, Substrate (chemistry), Environmental Science (miscellaneous), biology.organism_classification, Agricultural and Biological Sciences (miscellaneous), Pichia pastoris, chemistry.chemical_compound, Enzyme, Volume (thermodynamics), Bioreactor, Enzyme kinetics, Biotechnology |
| الوصف: | In this work, we statistically improved culture media for rPOXA 1B laccase production, expressed in Pichia pastoris containing pGAPZαA-LaccPost-Stop construct and assayed at 10 L bioreactor production scale (6 L effective work volume). The concentrated enzyme was evaluated for temperature and pH stability and kinetic parameter, characterized by monitoring oxidation of different ABTS [2, 20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] substrate concentrations. Plackett–Burman experimental design (PBED) implementation improved previous work results by 3.05-fold, obtaining a laccase activity of 1373.72 ± 0.37 U L−1 at 168 h of culture in a 500 mL shake flask. In contrast, one factor experimental design (OFED) applied after PBED improved by threefold the previous study, additionally increasing the C/N ratio. Employing OFED media at 10 L bioreactor scale was capable of producing 3159.93 ± 498.90 U L−1 at 192 h, representing a 2.4-fold increase. rPOXA 1B concentrate remained stable between 10 and 50 °C and retained over 70% residual enzymatic activity at 60 °C and 50% at 70 °C. Concerning pH stability, the enzyme was stable at pH 4.0 ± 0.2 with a residual activity greater than 90%. The lowest residual activity (60%) was obtained at pH 10.0 ± 0.2. Furthermore, the apparent kinetic parameters were Vmax of 3.163 × 10−2 mM min−1 and Km of 1.716 mM. Collectively, regarding enzyme stability our data provide possibilities for applications involving a wide range of pH and temperatures. |
| تدمد: | 2190-5738 2190-572X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::e15e16ef49aabcf50205128f4740e8b9 https://doi.org/10.1007/s13205-019-1979-y |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi...........e15e16ef49aabcf50205128f4740e8b9 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 21905738 2190572X |
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