ArticleAdvanced pharmaceutical bulletin2014
Optimized condition for enhanced soluble-expression of recombinant mutant anabaena variabilis phenylalanine ammonia lyase.
Article in Advanced pharmaceutical bulletin, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 14 citations in OpenAlex.
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- Cytoplasmic Chaperones Enhance Soluble Expression of Anti-EGFR huscFv inIranian journal of biotechnology · 2020Article
- Anti-HER2 scFv Expression in Escherichia coli SHuffleMolecular biotechnology · 2020Article
- Differentially expressed genes in heads and tails of Angelica sinensis diels: Focusing on ferulic acid metabolism.Chinese journal of integrative medicine · 2017Article
- Effect of Culture Condition Variables on Human Endostatin Gene Expression inJundishapur journal of microbiology · 2016Article
- Chaperone-Assisted Soluble Expression of a Humanized Anti-EGFR ScFv Antibody in E. Coli.Advanced pharmaceutical bulletin · 2015Article
- Effects of Environmental Factors on Soluble Expression of a Humanized Anti-TNF-α scFv Antibody in Escherichia coli.Advanced pharmaceutical bulletin · 2015Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
purposeRecently discovered Anabaena variabilis phenylalanine ammonia lyase (AvPAL) proved to be a good candidate for enzyme replacement therapy of phenylketonuria. Outstanding stability properties of a mutant version of this enzyme, produced already in our laboratory, have led us to the idea of culture conditions optimization for soluble expression of this therapeutically valuable enzyme in E. coli.
methodsIn the present study, the gene encoding mutant version of AvPAL was cloned into the pET28a expression vector. Different concentrations of IPTG, induction period, growth temperature, shaking speed, as well as different types of culture media were examined with respect to the amount of recombinant protein produced and specific activity of the enzyme.
resultsBased upon our findings, maximum amount of active mutant enzyme was attained by addition of 0.5 mM IPTG at 150 rpm to the TB culture media. The yield of active enzyme at cluture tempreature of 25 °C and induction period of 18 hour was the highest.
conclusionThe results of this study indicated that the yield of mutant AvPAL production in E. coli can be affected mainly by culture temperature and inducer concentration.
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