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Secretory expression and enzymatic characterization of recombinant Agarivorans albus β-agarase in Escherichia coli
- Source :
- Preparative Biochemistry & Biotechnology. 47:1037-1042
- Publication Year :
- 2017
- Publisher :
- Informa UK Limited, 2017.
-
Abstract
- Agarase catalyzes the hydrolysis of agar, which is primarily used as a medium for microbiology, various food additives, and new biomass materials. In this study, we described the expression of the synthetic gene encoding β-agarase from Agarivorans albus (Aaβ-agarase) in Escherichia coli. The synthetic β-agarase gene was designed based on the biased codons of E. coli to optimize its expression and extracellular secretion in an active, soluble form. The synthesized agarase gene, including its signal sequence, was cloned into the pET-26 expression vector, and the pET-Aaβ-agarase plasmid was introduced into E. coli BL21-Star (DE3) cells. The E. coli transformants were cultured for high-yield secretion of recombinant Aaβ-agarase in Luria-Bertani broth containing 0.6 mM isopropyl β-D-1-thiogalactopyranoside for 9 h at 37°C. The expressed recombinant Aaβ-agarase was purified by ammonium sulfate precipitation and diethylaminoethyl-sepharose column chromatography, yielding ∼10 mg/L Aaβ-agarase. The purified recombinant Aaβ-agarase exhibited optimal activity at pH 7 and 40°C, and its activity was strongly inhibited by Cu
- Subjects :
- 0301 basic medicine
food.ingredient
Glycoside Hydrolases
Genetic Vectors
030106 microbiology
Biology
medicine.disease_cause
Biochemistry
Microbiology
law.invention
03 medical and health sciences
Hydrolysis
Transformation, Genetic
food
law
Escherichia coli
medicine
Agar
Cloning, Molecular
Agarivorans albus
chemistry.chemical_classification
Alteromonadaceae
Food additive
Agarase
Temperature
General Medicine
Hydrogen-Ion Concentration
Recombinant Proteins
Kinetics
030104 developmental biology
Enzyme
chemistry
Recombinant DNA
biology.protein
Electrophoresis, Polyacrylamide Gel
Biotechnology
Subjects
Details
- ISSN :
- 15322297 and 10826068
- Volume :
- 47
- Database :
- OpenAIRE
- Journal :
- Preparative Biochemistry & Biotechnology
- Accession number :
- edsair.doi.dedup.....80f047a5987127928a578e7e2ce107ca
- Full Text :
- https://doi.org/10.1080/10826068.2017.1373292