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Effective lipase purification from Bacillus cereus NC7401: a novel approach using an alcohol-/salt-based aqueous two-phase system

Kulsoom Akhter, Eena Sadaf, Azeem Bibi, Jahanzeb Ayyub, Tahseen Ghous, and Tasleem Akhtar

Department of Chemistry, The University of Azad Jammu and Kashmir, Muzaffarabad, Pakistan



Received: 27 June 2023  Accepted: 1 October 2023


Alcohol-/salt-based aqueous two-phase system was utilized for the purification of lipase extracted from Bacillus cereus strains isolated from Tagetes minuta root soil contaminated with waste oil from automobile repairs. Tagetes minuta is known for its allelopathic properties which bear a variety of microbial communities in its root soil including bacteria with the potential to produce lipases with distinct structures and activities. Exploring these microbial strains may help to discover novel lipases with improved stability, substrate specificity and efficiency compared to already known enzymes. Automobile repair waste oil contains hydrocarbons and other contaminants while the presence of lipase-producing bacteria in the root soil of Tagetes minuta presents an opportunity for bioremediation, as lipases can play a vital role in breaking down complex hydrocarbons into more manageable compounds. Nine different systems consisting of alcohols and salts were investigated to study the efficiency of different concentrations of phase-forming constituents for lipase purification. The supreme lipase recovery was observed in an aqueous two-phase system with xylitol (45% w/w) and potassium phosphate (90% w/w) with a purification factor of 24.14 and yield of 87.71%. The highest activity of the purified lipase was obtained at a temperature of 40 °C and pH 7.0. The present study revealed that this revolutionary aqueous two-phase technique can be utilized to purify and extract important enzymes more efficiently and cost-effectively.

Keywords: Lipase; Aqueous two-phase system; Bacillus cereus; Tagetes minuta

Full paper is available at

DOI: 10.1007/s11696-023-03133-3


Chemical Papers 78 (2) 963–971 (2024)

Monday, April 22, 2024

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