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Partitioning behavior of recombinant lipase in Escherichia coli by ionic liquid-based aqueous two-phase systems

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논문

Partitioning behavior of recombinant lipase in Escherichia coli by ionic liquid-based aqueous two-phase systems

학술지

RSC advances

저자명

Hadzir, Muhammad Hakimi; Abbasiliasi, Sahar; Ariff, Arbakariya B.; Yusoff, Siti Baidurah; Ng, Hui Suan; Tan, Joo Shun

초록

<P>There has been significant interest in ionic liquid aqueous two-phase systems (ILATPSs) with properties such as rapid phase segregation which can lead to a reduction in time taken for protein recovery. Evaluations of ILATPSs were performed with various types of ionic liquid ((Emim)BF4, (Emim)Br, (Bmim)BF4, and (Bmim)Br) and salts (potassium-, sodium-, and magnesium-based) as phase components to figure out their competencies in the recovery of lipase from a fermentation broth of <I>E. coli</I> using banana waste as a substrate. The results of this study revealed that an ILATPS comprising (Emim)Br/potassium phosphate significantly enhanced lipase recovery upon partitioning lipase. Optimization of the composition of the ILATPS using response surface methodology (RSM) significantly improved the purification factor (<I>P</I>F) and partition coefficient (<I>K</I>e). The influences of crude loading (<I>C</I>L), pH changes, and the presence of NaCl on the recovery performance were also studied. Recovery of <I>E. coli</I> BL21 lipase was accomplished in an (Emim)Br/potassium phosphate ILATPS using 26.5% (w/w) (Emim)Br, 19% (w/w) potassium phosphate at pH 7.6, 3% NaCl and a crude loading of 7% (w/w). Using this ILATPS, lipase was successfully recovered in a single purification step, which gave a yield, <I>P</I>F and <I>K</I>e of 93.75%, 3.394 and 1.352, respectively. A high <I>P</I>F value indicates that (Emim)Br/potassium phosphate is capable of attaining an excellent degree of lipase purity, suggesting that the proposed ILATPS is suitable for implementation in a large scale process for lipase purification.</P>

발행연도

2016

발행기관

The Royal Society of Chemistry

ISSN

2046-2069

6

86

페이지

pp.82571-82580

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1 2023-12-11

논문; 2016-01-01

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