Abstract
Recombinantly produced enzymes are applied in many fields, ranging from medicine to food and nutrition, production of detergents, textile, leather, paper, pulp, and plastics. Thus, the cost-effectiveness of recombinant enzyme synthesis is an important issue in biotechnological industry. Isopropyl-β-D-thiogalactoside (IPTG), an analog of lactose, is currently the most widely used chemical agent for the induction of recombinant enzyme synthesis. However, the use of IPTG can lead to production of toxic elements and can introduce physiological stress to cells. Thus, this study aims to find a simpler, cheaper, and safer way to produce recombinant enzymes. In this study, production of several previously designed recombinant lipolytic enzymes (GDEst-95 esterase, GD-95RM lipase, fused GDEst-lip lipolytic enzyme, and putative cutinase Cut+SP from Streptomyces scabiei 87.22) is induced in E. coli BL21 (DE3) using 4 mM milk permeate, a type of waste of the milk manufacturing process possessing >82% lactose. The SDS-PAGE analysis clearly indicates synthesis of all target enzymes during a 2–12 h post-induction timeframe. Further investigation of GDEst-95, GD-95RM, GDEst-lip, and Cut+SP biocatalysts was carried out spectrophotometrically and using zymography method, confirming production of fully active enzymes.
Funder
Research Council of Lithuania
Subject
Plant Science,Biochemistry, Genetics and Molecular Biology (miscellaneous),Food Science
Reference96 articles.
1. Demain, A.L., and Vaishnav, P. (2016). Reference Module in Food Science, Elsevier.
2. How to achieve high-level expression of microbial enzymes;Liu;Bioengineered,2013
3. E. coli HMS174(DE3) is a sustainable alternative to BL21(DE3);Hausjell;Microb. Cell Factories,2018
4. Strain engineering for improved expression of recombinant proteins in bacteria;Makino;Microb. Cell Factories,2011
5. Critical Factors Affecting the Success of Cloning, Expression, and Mass Production of Enzymes by Recombinant E. coli.;Fakruddin;ISRN Biotechnol.,2013
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