» Articles » PMID: 30962822

Improving Squalene Production by Enhancing the NADPH/NADP Ratio, Modifying the Isoprenoid-feeding Module and Blocking the Menaquinone Pathway in

Overview
Publisher Biomed Central
Specialty Biotechnology
Date 2019 Apr 10
PMID 30962822
Citations 17
Authors
Affiliations
Soon will be listed here.
Abstract

Background: Squalene is currently used widely in the food, cosmetics, and medicine industries. It could also replace petroleum as a raw material for fuels. Microbial fermentation processes for squalene production have been emerging over recent years. In this study, to study the squalene-producing potential of (, we employed several increasing strategies for systematic metabolic engineering. These include the expression of human truncated squalene synthase, the overexpression of rate-limiting enzymes in isoprenoid pathway, the modification of isoprenoid-feeding module and the blocking of menaquinone pathway.

Results: Herein, human truncated squalene synthase was engineered in to create a squalene-producing bacterial strain. To increase squalene yield, we employed several metabolic engineering strategies. A fivefold squalene titer increase was achieved by expressing rate-limiting enzymes (IDI, DXS, and FPS) involved in the isoprenoid pathway. Pyridine nucleotide transhydrogenase (UdhA) was then expressed to improve the cellular NADPH/NADP ratio, resulting in a 59% increase in squalene titer. The Embden-Meyerhof pathway (EMP) was replaced with the Entner-Doudoroff pathway (EDP) and pentose phosphate pathway (PPP) to feed the isoprenoid pathway, along with the overexpression of and genes which encode rate-limiting enzymes in the EDP and PPP, leading to a 104% squalene content increase. Based on the blocking of menaquinone pathway, a further 17.7% increase in squalene content was achieved. Squalene content reached a final 28.5 mg/g DCW and 52.1 mg/L.

Conclusions: This study provided novel strategies for improving squalene yield and demonstrated the potential of producing squalene by .

Citing Articles

Unveiling the Synergistic Effect of Salicylic Acid on Triterpenoid Biosynthesis in : Elucidating the Molecular Underpinnings.

Hu F, Fang Y, Khan Z, Xing L Int J Mol Sci. 2025; 26(3).

PMID: 39940765 PMC: 11816812. DOI: 10.3390/ijms26030996.


Microbial Squalene: A Sustainable Alternative for the Cosmetics and Pharmaceutical Industry - A Review.

Shalu S, Karthikanath P, Vaidyanathan V, Blank L, Germer A, Balakumaran P Eng Life Sci. 2024; 24(10):e202400003.

PMID: 39391272 PMC: 11464149. DOI: 10.1002/elsc.202400003.


Identification of a novel NADPH generation reaction in the pentose phosphate pathway in using mBFP.

Ueno K, Sawada S, Ishibashi M, Kanda Y, Shimizu H, Toya Y J Bacteriol. 2024; 206(11):e0027624.

PMID: 39387572 PMC: 11580446. DOI: 10.1128/jb.00276-24.


Identification of a novel metabolic engineering target for carotenoid production in Saccharomyces cerevisiae via ethanol-induced adaptive laboratory evolution.

Su B, Li A, Deng M, Zhu H Bioresour Bioprocess. 2024; 8(1):47.

PMID: 38650275 PMC: 10992865. DOI: 10.1186/s40643-021-00402-5.


Enhancing menaquinone-7 biosynthesis by adaptive evolution of Bacillus natto through chemical modulator.

Zhang B, Peng C, Lu J, Hu X, Ren L Bioresour Bioprocess. 2024; 9(1):120.

PMID: 38647796 PMC: 10992315. DOI: 10.1186/s40643-022-00609-0.


References
1.
Verho R, Londesborough J, Penttila M, Richard P . Engineering redox cofactor regeneration for improved pentose fermentation in Saccharomyces cerevisiae. Appl Environ Microbiol. 2003; 69(10):5892-7. PMC: 201209. DOI: 10.1128/AEM.69.10.5892-5897.2003. View

2.
Ghimire G, Thuan N, Koirala N, Sohng J . Advances in Biochemistry and Microbial Production of Squalene and Its Derivatives. J Microbiol Biotechnol. 2015; 26(3):441-51. DOI: 10.4014/jmb.1510.10039. View

3.
Lewis T, Nichols P, McMEEKIN T . Sterol and squalene content of a docosahexaenoic-acid-producing thraustochytrid: influence of culture age, temperature, and dissolved oxygen. Mar Biotechnol (NY). 2004; 3(5):439-47. DOI: 10.1007/s10126-001-0016-3. View

4.
Unden G . Differential roles for menaquinone and demethylmenaquinone in anaerobic electron transport of E. coli and their fnr-independent expression. Arch Microbiol. 1988; 150(5):499-503. DOI: 10.1007/BF00422294. View

5.
Chen G, Fan K, Lu F, Li Q, Aki T, Chen F . Optimization of nitrogen source for enhanced production of squalene from thraustochytrid Aurantiochytrium sp. N Biotechnol. 2010; 27(4):382-9. DOI: 10.1016/j.nbt.2010.04.005. View