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Original article
Improvement and Prediction of Secondary Metabolites Biosynthesis Under Yeast Extract Elicitation of Neem Cell Suspension Culture Using Response Surface Methodology
Ghasemali Garoosi
Imam Khomeini International University
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9144-7585
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Neem is the medicinal plant used as antimalarial, antibacterial, antiviral, insecticide and antimicrobial. The aim of this study was investigate effect of yeast extract and sampling time on cell growth, secondary metabolites synthesis, SQS1 and MOF1 genes expression and prediction of secondary metabolites synthesis by response surface methodology. Highest fresh and dry cell weight were 580.25 g/L and 21.01 g/L obtained 6 days after addition 100 mg/L yeast extract. Highest azadirachtin accumulation and production were 16.08 mg/g DW and 219.78 mg/L obtained at 25 mg/L yeast extract for 2 and 4 days. Maximum mevalonic acid accumulation and production were 1.746 mg/g DW and 23.77 mg/L observed 2 days after addition 50 mg/L yeast extract addition. Highest amount of squalene accumulation (0.215 mg/g DW) and production (4.526 mg/L) were achieved 4 days after application of 50 mg/L yeast extract. Prediction results showed that highest azadirachtin accumulation (13.607 mg/g DW) and production (190.50 mg/L), mevalonic acid accumulation (0.500 mg/g DW) and production (5.57 mg/L) and squalene accumulation (0.301 mg/g DW) obtained by using 245 mg/L yeast extract for 2 days, 71 mg/L yeast extract for 2 days, 200 mg/L yeast extract for 4.96 days, whitout yeast extract for 6.54 days and 4 days, respectively and predicted that highest squalene production is achieved by long-term exposure to high concentrations of yeast extract. qRT-PCR analysis showed the maximum relative gene expression of SQS1 and MOF1 by addition 150 mg/L yeast extract for 4 days and 25 mg/L yeast extract for 2 days, respectively.
Keywords
Azadiractin, Mevalonic acid, Neem, Response surface methodology, Squalene, Yeast extract
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This preprint is under consideration at AMB Express. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Original article
Improvement and Prediction of Secondary Metabolites Biosynthesis Under Yeast Extract Elicitation of Neem Cell Suspension Culture Using Response Surface Methodology
Ghasemali Garoosi
Imam Khomeini International University
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9144-7585
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Revision
Version 1
Posted 08 Jan, 2021
No community comments so far
Editorial decision: Major Revision
On 13 Jan, 2021
Review #2 received
Received 11 Jan, 2021
Review #1 received
Received 11 Jan, 2021
Reviewer #2 agreed
On 05 Jan, 2021
Reviewer #1 agreed
On 02 Jan, 2021
Reviewers invited
Invitations sent on 01 Jan, 2021
Editor assigned
On 29 Dec, 2020
Submission checks complete
On 29 Dec, 2020
Editor invited
On 29 Dec, 2020
First submitted
On 27 Dec, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Neem is the medicinal plant used as antimalarial, antibacterial, antiviral, insecticide and antimicrobial. The aim of this study was investigate effect of yeast extract and sampling time on cell growth, secondary metabolites synthesis, SQS1 and MOF1 genes expression and prediction of secondary metabolites synthesis by response surface methodology. Highest fresh and dry cell weight were 580.25 g/L and 21.01 g/L obtained 6 days after addition 100 mg/L yeast extract. Highest azadirachtin accumulation and production were 16.08 mg/g DW and 219.78 mg/L obtained at 25 mg/L yeast extract for 2 and 4 days. Maximum mevalonic acid accumulation and production were 1.746 mg/g DW and 23.77 mg/L observed 2 days after addition 50 mg/L yeast extract addition. Highest amount of squalene accumulation (0.215 mg/g DW) and production (4.526 mg/L) were achieved 4 days after application of 50 mg/L yeast extract. Prediction results showed that highest azadirachtin accumulation (13.607 mg/g DW) and production (190.50 mg/L), mevalonic acid accumulation (0.500 mg/g DW) and production (5.57 mg/L) and squalene accumulation (0.301 mg/g DW) obtained by using 245 mg/L yeast extract for 2 days, 71 mg/L yeast extract for 2 days, 200 mg/L yeast extract for 4.96 days, whitout yeast extract for 6.54 days and 4 days, respectively and predicted that highest squalene production is achieved by long-term exposure to high concentrations of yeast extract. qRT-PCR analysis showed the maximum relative gene expression of SQS1 and MOF1 by addition 150 mg/L yeast extract for 4 days and 25 mg/L yeast extract for 2 days, respectively.
Figure 1
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Figure 4