Preparation of stable cross-linked enzyme aggregates (CLEAs) of a Ureibacillus thermosphaericus esterase for application in malathion removal from wastewater

Yuliya V. Samoylova, Ksenia N. Sorokina, Alexander V. Piligaev, Valentin N. Parmon

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16 Citations (Scopus)

Abstract

In this study, the active and stable cross-linked enzyme aggregates (CLEAs) of the thermostable esterase estUT1 of the bacterium Ureibacillus thermosphaericus were prepared for application in malathion removal from municipal wastewater. Co-expression of esterase with an E. coli chaperone team (KJE, ClpB, and ELS) increased the activity of the soluble enzyme fraction up to 200.7 ± 15.5 U mg–1. Response surface methodology (RSM) was used to optimize the preparation of the CLEA-estUT1 biocatalyst to maximize its activity and minimize enzyme loss. CLEA-estUT1 with the highest activity of 29.4 ± 0.5 U mg–1 (90.6 ± 2.7% of the recovered activity) was prepared with 65.1% (w/v) ammonium sulfate, 120.6 mM glutaraldehyde, and 0.2 mM bovine serum albumin at 5.1 h of cross-linking. The biocatalyst has maximal activity at 80 ºC and pH 8.0. Analysis of the properties of CLEA-estUT1 and free enzyme at 50–80 ºC and pH 5.0–10.0 showed higher stability of the biocatalyst. CLEA-estUT1 showed marked tolerance against a number of chemicals and high operational stability and activity in the reaction of malathion hydrolysis in wastewater (up to 99.5 ± 1.4%). After 25 cycles of malathion hydrolysis at 37 ºC, it retained 55.2 ± 1.1% of the initial activity. The high stability and reusability of CLEA-estUT1 make it applicable for the degradation of insecticides.

Original languageEnglish
Article number154
Number of pages19
JournalCatalysts
Volume8
Issue number4
DOIs
Publication statusPublished - 11 Apr 2018

Keywords

  • Cross-linked enzyme aggregates
  • Esterase
  • Malathion hydrolysis
  • Molecular chaperones
  • Response surface methodology
  • Ureibacillus thermosphaericus
  • BIODEGRADATION
  • MOLECULAR CHAPERONES
  • ESCHERICHIA-COLI
  • response surface methodology
  • BACTERIAL RECOMBINANT LIPASE
  • IDENTIFICATION
  • IMMOBILIZATION
  • cross-linked enzyme aggregates
  • FATTY-ACID
  • malathion hydrolysis
  • esterase
  • molecular chaperones
  • PROTEINS
  • EXPRESSION
  • THERMAL INACTIVATION

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