Partition separation and characterization of the polyhydroxyalkanoates synthase produced from recombinant Escherichia coli using an aqueous two-phase system

Natural products span broad activities and applications; however, their access and production are often limited by native cellular sources. As a result, the heterologous production of a siderophore termed yersiniabactin (Ybt) was completed using the surrogate host Escherichia coli. Post-production and purification steps are complicated by the complex nature of most media used for cell growth, prompting the development in this work of an aqueous two-phase pre-purification system capable of rapidly and simply enhancing the concentration of the target Ybt compound.

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Purification and characterization of a collagenase from Penicillium sp. UCP 1286 by polyethylene glycol-phosphate aqueous two-phase system

Protein Expression and Purification ◽

10.1016/j.pep.2017.02.010 ◽

2017 ◽

Vol 133 ◽

pp. 8-14 ◽

Cited By ~ 14

Author(s):

Maria Carolina de Albuquerque Wanderley ◽

José Manoel Wanderley Duarte Neto ◽

Wendell Wagner Campos Albuquerque ◽

Daniela de Araújo Viana Marques ◽

Carolina de Albuquerque Lima ◽

...

Keyword(s):

Polyethylene Glycol ◽

Phase System ◽

Purification And Characterization ◽

Two Phase ◽

Aqueous Two Phase System ◽

Penicillium Sp

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Production of styrene oxide from styrene by a recombinant Escherichia coli with enhanced AcrAB-TolC efflux pump level in an aqueous-organic solvent two-phase system

Bioscience Biotechnology and Biochemistry ◽

10.1080/09168451.2020.1755219 ◽

2020 ◽

Vol 84 (7) ◽

pp. 1513-1520

Author(s):

Noriyuki Doukyu ◽

Shinichiro Iida

Keyword(s):

Escherichia Coli ◽

Organic Solvent ◽

Efflux Pump ◽

Styrene Oxide ◽

Recombinant Escherichia Coli ◽

Phase System ◽

Pump Level ◽

Two Phase

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One-step purification of histone deacetylase from Escherichia coli cell-lysate by counter-current chromatography using aqueous two-phase system

Journal of Chromatography A ◽

10.1016/j.chroma.2007.01.111 ◽

2007 ◽

Vol 1151 (1-2) ◽

pp. 158-163 ◽

Cited By ~ 5

Author(s):

Yoichi Shibusawa ◽

Naoko Takeuchi ◽

Kanako Tsutsumi ◽

Shigeru Nakano ◽

Akio Yanagida ◽

...

Keyword(s):

Escherichia Coli ◽

Histone Deacetylase ◽

Coli Cell ◽

Phase System ◽

Two Phase ◽

Aqueous Two Phase System ◽

Cell Lysate ◽

One Step ◽

Counter Current

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Extraction of plasmid DNA from Escherichia coli cell lysate in a thermoseparating aqueous two-phase system

Journal of Chromatography A ◽

10.1016/j.chroma.2003.10.028 ◽

2004 ◽

Vol 1024 (1-2) ◽

pp. 95-104 ◽

Cited By ~ 53

Author(s):

Cecilia Kepka ◽

Jenny Rhodin ◽

Raf Lemmens ◽

Folke Tjerneld ◽

Per-Erik Gustavsson

Keyword(s):

Escherichia Coli ◽

Plasmid Dna ◽

Coli Cell ◽

Phase System ◽

Two Phase ◽

Aqueous Two Phase System ◽

Cell Lysate

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Purification and Characterization of Lipase Produced by Leuconostoc mesenteroides Subsp. mesenteroides ATCC 8293 Using an Aqueous Two-Phase System (ATPS) Composed of Triton X-100 and Maltitol

Molecules ◽

10.3390/molecules23071800 ◽

2018 ◽

Vol 23 (7) ◽

pp. 1800 ◽

Cited By ~ 2

Author(s):

Nurfadhilah Eko Sukohidayat ◽

Mohammad Zarei ◽

Badlishah Baharin ◽

Mohd Manap

Keyword(s):

Phase System ◽

Two Phase ◽

Aqueous Two Phase System ◽

Process Characteristics ◽

Sds Page ◽

Ph Range ◽

Lipase Purification ◽

Triton X ◽

First Time

Purification of lipase produced by L. mesenteroides subsp. mesenteroides ATCC 8293 was conducted for the first time using a novel aqueous two-phase system (ATPS) composed of Triton X-100 and maltitol. The partitioning of lipase was optimized according to several parameters including pH, temperature, and crude load. Results showed that lipase preferentially migrated to the Triton X-100 rich phase and optimum lipase partitioning was achieved in ATPS at TLL of 46.4% and crude load of 20% at 30 °C and pH 8, resulting in high lipase purification factor of 17.28 and yield of 94.7%. The purified lipase showed a prominent band on SDS-PAGE with an estimated molecular weight of 50 kDa. The lipase was stable at the temperature range of 30–60 °C and pH range of 6–11, however, it revealed its optimum activity at the temperature of 37 °C and pH 8. Moreover, lipase exhibited enhanced activity in the presence of non-ionic surfactants with increased activity up to 40%. Furthermore, results exhibited that metals ions such as Na+, Mg2+, K+ and Ca2+ stimulated lipase activity. This study demonstrated that this novel system could be potentially used as an alternative to traditional ATPS for the purification and recovery of enzymes since the purified lipase still possesses good process characteristics after undergoing the purification process.

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