ArticleCurrent microbiology2009
Expression of methyl parathion hydrolase in Pichia pastoris.
Article in Current microbiology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
4 citing papers in PubMed, 29 citations in OpenAlex.
- Enhanced Secretory Expression and Surface Display Level of Bombyx mori Acetylcholinesterase 2 by Pichia pastoris Based on Codon Optimization Strategy for Pesticides Setection.Applied biochemistry and biotechnology · 2021Article
- Enhanced secretion of a methyl parathion hydrolase in Pichia pastoris using a combinational strategy.Microbial cell factories · 2015Article
- Enhanced production of Thermomyces lanuginosus lipase in Pichia pastoris via genetic and fermentation strategies.Journal of industrial microbiology & biotechnology · 2014Article
- Improving the secretion of a methyl parathion hydrolase in Pichia pastoris by modifying its N-terminal sequence.PloS one · 2014Article
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In the Pichia pastoris expression system, increasing the copy number of the expression cassette often has the effect of increasing the amount of protein expressed. To improve the expression level of methyl parathion hydrolase (MPH), we constructed two integration vectors with four and eight direct repeats of the expression cassette using an in vitro multimerization approach. After two successive integrations, at least 12 copies of the MPH expression cassette were integrated into the P. pastoris chromosome. Under shake-flask conditions, over 55 mg active MPH/l was secreted into the medium by the multicopy clones. The extracellular enzyme activity was about 10-fold higher for the multicopy clones than for clones containing a single copy of the gene. Further investigations revealed that the multicopy MPH expression cassette could remain stably integrated and functional over five generations. Note that the expression vector pRF constructed in our study can be not only used to construct multiple copies of the expression cassette in vitro, but also integrated into the P. pastoris genome without introducing any antibiotic resistance gene, which is desirable for production of biotherapeutic proteins.
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Registered trials
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