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Enhancing production of ergosterol in Pichia pastoris GS115 by over-expression of 3-hydroxy-3-methylglutaryl CoA reductase from Glycyrrhiza uralensis
- Source :
- Acta Pharmaceutica Sinica. B, Acta Pharmaceutica Sinica B, Vol 4, Iss 2, Pp 161-166 (2014), Acta Pharmaceutica Sinica B
- Publication Year :
- 2014
- Publisher :
- Elsevier, 2014.
-
Abstract
- The rate-limiting enzyme in the mevalonic acid (MVA) pathway which can lead to triterpenoid saponin glycyrrhizic acid (GA) is 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR). In order to reveal the effect of copy number variation in the HMGR gene on the MVA pathway, the HMGR gene from Glycyrrhiza uralensis Fisch. (GuHMGR) was cloned and over-expressed in Pichia pastoris GS115. Six recombinant P. pastoris strains containing different copy numbers of the GuHMGR gene were obtained and the content of ergosterol was analyzed by HPLC. The results showed that all the recombinant P. pastoris strains contained more ergosterol than the negative control and the strains with 8 and 44 copies contained significantly more ergosterol than the other strains. However, as the copy number increased, the content of ergosterol showed an increasing–decreasing–increasing pattern. This study provides a rationale for increasing the content of GA through over-expressing the GuHMGR gene in cultivars of G. uralensis.<br />Graphical abstract Six recombinant Pichia pastoris strains containing different copy numbers of the GuHMGR gene were obtained in this paper. HPLC analysis showed that the content of ergosterol in all recombinant P. pastoris strains containing the GuHMGR gene was higher than in negative control. And with the increase of copy number of the GuHMGR gene, the content of ergosterol was presented in an increasing–decreasing–increasing pattern.
- Subjects :
- LLOQ, lower limit of quantitation
BMMY, buffered methanol-complex medium
CNV, copy number variation
BMGY, buffered glycerol-complex medium
RSD, relative standard deviation
Mevalonic acid
Reductase
MM, minimal medium
law.invention
Pichia pastoris
HMGR, 3-hydroxy-3-methylglutaryl-CoA reductase
chemistry.chemical_compound
PCR, polymerase chain reaction
law
Botany
Glycyrrhiza uralensis Fisch
MD, minimal dextrose medium
General Pharmacology, Toxicology and Pharmaceutics
Triterpenoid saponin
chemistry.chemical_classification
Ergosterol
LOD, limit of detection
biology
Copy number variation
lcsh:RM1-950
Glycyrrhiza uralensis
biology.organism_classification
3-Hydroxy-3-methylglutaryl-CoA reductase gene
lcsh:Therapeutics. Pharmacology
Enzyme
chemistry
Biochemistry
Over-expression
Recombinant DNA
Original Article
MVA, mevalonic acid
YPD, yeast peptone dextrose medium
Subjects
Details
- Language :
- English
- ISSN :
- 22113843 and 22113835
- Volume :
- 4
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Acta Pharmaceutica Sinica. B
- Accession number :
- edsair.doi.dedup.....8c5348d45e52d539b5873889e1a9634b