Alternative Heterologous Expression of L-Arabinose Isomerase from Enterococcus faecium DBFIQ E36 By Residual Whey Lactose Induction

Autor: Denise Cavalcante Hissa, Luciana Rocha Barros Gonçalves, Ticiane C. de Souza, Ricardo Martín Manzo, Enrique José Mammarella, Saulo GonÇalves de Santiago Bezerra, Ravena Casemiro Oliveira
Rok vydání: 2021
Předmět:
0106 biological sciences
Glycerol
Isopropyl Thiogalactoside
Enterococcus faecium
lac operon
Bioengineering
Lactose
L-arabinose isomerase
01 natural sciences
Applied Microbiology and Biotechnology
Biochemistry
Gene Expression Regulation
Enzymologic
l-Arabinose isomerase
03 medical and health sciences
chemistry.chemical_compound
Affinity chromatography
Bacterial Proteins
010608 biotechnology
Whey
Escherichia coli
Inducer
Food science
Cheese whey
Cloning
Molecular
Molecular Biology
Aldose-Ketose Isomerases
030304 developmental biology
chemistry.chemical_classification
0303 health sciences
biology
d-Tagatose
Gene Expression Regulation
Bacterial
Auto-induction
Hydrogen-Ion Concentration
biology.organism_classification
Recombinant Proteins
Culture Media
Enzyme
Glucose
chemistry
Heterologous expression
· Escherichia coli expression
Biotechnology
Zdroj: Repositório Institucional da Universidade Federal do Ceará (UFC)
Universidade Federal do Ceará (UFC)
instacron:UFC
ISSN: 1559-0305
Popis: This study reports an alternative strategy for the expression of a recombinant L-AI from Enterococcus faecium DBFIQ E36 by auto-induction using glucose and glycerol as carbon sources and residual whey lactose as inducer agent. Commercial lactose and isopropyl β-D-1-thiogalactopyranoside (IPTG) were also evaluated as inducers for comparison of enzyme expression levels. The enzymatic extracts were purified by affinity chromatography, characterized, and applied in the bioconversion of D-galactose into D-tagatose. L-AI presented a catalytic activity of 1.67 ± 0.14, 1.52 ± 0.01, and 0.7 ± 0.04 U/mL, when expressed using commercial lactose, lactose from whey, and IPTG, respectively. Higher activities could be obtained by changing the protocol of enzyme extraction and, for instance, the enzymatic extract produced with whey presented a catalytic activity of 3.8 U/mL. The specific activity of the enzyme extracts produced using lactose (commercial or residual whey) after enzyme purification was also higher when compared to the enzyme expressed with IPTG. Best results were achieved when enzyme expression was conducted using 4 g/L of residual whey lactose for 11 h. These results proved the efficacy of an alternative and economic protocol for the effective expression of a recombinant L-AI aiming its high-scale production.
Databáze: OpenAIRE
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