TY - JOUR

T1 - Preparation of stable cross-linked enzyme aggregates (CLEAs) of a Ureibacillus thermosphaericus esterase for application in malathion removal from wastewater

AU - Samoylova, Yuliya V.

AU - Sorokina, Ksenia N.

AU - Piligaev, Alexander V.

AU - Parmon, Valentin N.

N1 - Publisher Copyright: © 2018 by the authors.

PY - 2018/4/11

Y1 - 2018/4/11

N2 - 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.

AB - 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.

KW - Cross-linked enzyme aggregates

KW - Esterase

KW - Malathion hydrolysis

KW - Molecular chaperones

KW - Response surface methodology

KW - Ureibacillus thermosphaericus

KW - BIODEGRADATION

KW - MOLECULAR CHAPERONES

KW - ESCHERICHIA-COLI

KW - response surface methodology

KW - BACTERIAL RECOMBINANT LIPASE

KW - IDENTIFICATION

KW - IMMOBILIZATION

KW - cross-linked enzyme aggregates

KW - FATTY-ACID

KW - malathion hydrolysis

KW - esterase

KW - molecular chaperones

KW - PROTEINS

KW - EXPRESSION

KW - THERMAL INACTIVATION

UR - http://www.scopus.com/inward/record.url?scp=85045387811&partnerID=8YFLogxK

U2 - 10.3390/catal8040154

DO - 10.3390/catal8040154

M3 - Article

AN - SCOPUS:85045387811

VL - 8

JO - Catalysts

JF - Catalysts

SN - 2073-4344

IS - 4

M1 - 154

ER -