High-level recombinant production of squalene using selected<i>Saccharomyces cerevisiae</i>strains

Han, Jong Yun; Seo, Sung Hwa; Song, Jae Myeong; Lee, Hongweon; Choi, Eui‐Sung

doi:10.1007/s10295-018-2018-4

Cited by 43 publications

(34 citation statements)

References 43 publications

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“…(A) Squalene biosynthesis via MVA pathway in yeast, fungi, and algae. The engineering strategies for enhanced squalene production are as follows: overexpression of HMGR (Polakowski et al, 1998; Tokuhiro et al, 2009; Mantzouridou and Tsimidou, 2010; Dai et al, 2012, 2014; Zhuang and Chappell, 2015; Rasool et al, 2016a,b; Kwak et al, 2017; Paramasivan and Mutturi, 2017; Han et al, 2018; Huang et al, 2018; Wei et al, 2018) and SQS (Dai et al, 2014; Zhuang and Chappell, 2015; Rasool et al, 2016a,b), downregulation of SQE (Garaiová et al, 2014; Hull et al, 2014; Zhuang and Chappell, 2015; Rasool et al, 2016a,b; Han et al, 2018) in yeast; downregulation of SQE in algae (Kajikawa et al, 2015). (B) Squalene biosynthesis via MEP pathway in bacteria.…”

Section: Squalene Biosynthetic Pathway In Microorganismsmentioning

confidence: 99%

“…Their attempt to overexpress tHMGR and upc2.1 (a mutated regulatory factor that induces sterol biosynthetic gene) in S. cerevisiae BY4742 to yield miltiradiene in upcoming steps surprisingly led to over accumulation of squalene as an intermediate product. Very recently, Han et al (2018) overexpressed tHMG1 and ispA (bacterial FPP synthase) genes in S. cerevisiae Y2805 strain and obtained squalene concentration up to 400 ± 45 mg/L. They further increased squalene yield up to 756 ± 36 mg/L by the inclusion of terbinafine for partial inhibition of SQE.…”

Section: Engineering Of Microorganisms For Squalene Productionmentioning

confidence: 99%

“…Rapid and remarkable advances in genetic engineering, metabolic engineering and synthetic biology approaches over the last few decades have facilitated the insertion of heterologous genes and editing of the genome of an organism, enabling successful attempts to produce innumerable molecules of industrial importance (Martin et al, 2009; Stephanopoulos, 2012; Singh, 2014; Jullesson et al, 2015). Lately, a number of microorganisms have been engineered by inserting a squalene biosynthetic pathway or by the modification of existing biosynthetic pathway for over-production of squalene (Ghimire et al, 2009; Katabami et al, 2015;Han et al, 2018; Wei et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

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Engineering Strategies in Microorganisms for the Enhanced Production of Squalene: Advances, Challenges and Opportunities

Gohil

Bhattacharjee

Khambhati

et al. 2019

Front. Bioeng. Biotechnol.

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The triterpene squalene is a natural compound that has demonstrated an extraordinary diversity of uses in pharmaceutical, nutraceutical, and personal care industries. Emboldened by this range of uses, novel applications that can gain profit from the benefits of squalene as an additive or supplement are expanding, resulting in its increasing demand. Ever since its discovery, the primary source has been the deep-sea shark liver, although recent declines in their populations and justified animal conservation and protection regulations have encouraged researchers to identify a novel route for squalene biosynthesis. This renewed scientific interest has profited from immense developments in synthetic biology, which now allows fine-tuning of a wider range of plants, fungi, and microorganisms for improved squalene production. There are numerous naturally squalene producing species and strains; although they generally do not make commercially viable yields as primary shark liver sources can deliver. The recent advances made toward improving squalene output from natural and engineered species have inspired this review. Accordingly, it will cover in-depth knowledge offered by the studies of the natural sources, and various engineering-based strategies that have been used to drive the improvements in the pathways toward large-scale production. The wide uses of squalene are also discussed, including the notable developments in anti-cancer applications and in augmenting influenza vaccines for greater efficacy.

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Section: Squalene Biosynthetic Pathway In Microorganismsmentioning

confidence: 99%

Section: Engineering Of Microorganisms For Squalene Productionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Engineering Strategies in Microorganisms for the Enhanced Production of Squalene: Advances, Challenges and Opportunities

Gohil

Bhattacharjee

Khambhati

et al. 2019

Front. Bioeng. Biotechnol.

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“…It was consistent with our findings that medium C/N ratio was beneficial for squalene synthesis. By applying these engineering strategies, we have obtained an oleaginous yeast strain with a similar squalene level to the strain S. cerevisiae (Han, Seo, Song, Lee, & Choi, 2018;Huang et al, 2018). This work highlights the potential of engineering Y. lipolytica as a promising microbial platform for efficient synthesis of squalene and terpene-related compounds.…”

Section: Shake Flask Cultivation Of Engineered Strain With Ph and Carmentioning

confidence: 81%

Optimizing mevalonate pathway for squalene production inYarrowia lipolytica

Liu

Wang

Deng

et al. 2020

Preprint

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Li Deng) and pengxu@umbc.edu (PX). AbstractSqualene is the gateway molecule for triterpene-based natural products and steroidsbased pharmaceuticals. As a super lubricant, it has been used widely in health care industry due to its skin compatibility and thermostability. Squalene is traditionally sourced from sharkhunting or oil plant extraction, which is cost-prohibitive and not sustainable. Reconstitution of squalene biosynthetic pathway in microbial hosts is considered as a promising alternative for cost-efficient and scalable synthesis of squalene. In this work, we reported the engineering of the oleaginous yeast, Y. lipolytica, as a potential host for squalene production. We systematically identified the bottleneck of the pathway and discovered that the native HMG-CoA reductase led to the highest squalene improvement. With the recycling of NADPH from the mannitol cycle, the engineered strain produced about 180.3 mg/l and 188.2 mg/L squalene from glucose or acetate minimal media, respectively. By optimizing the C/N ratio, controlling the media pH and mitigating the acetyl-CoA flux competition from lipogenesis, the engineered strain produced about 502.7 mg/L squalene in shake flaks, a 28-fold increase compared to the parental strain (17.2 mg/L). We also profiled the metabolic byproducts citric acid and mannitol level and observed that they are reincorporated into cell metabolism at the late stage of fermentation. This work may serve as a baseline to harness Y. lipolytica as an oleaginous cell factory for production of squalene or terpene-based chemicals.

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“…Overexpression of HMG has been shown to boost the production of α-farnesene, linalool and limonene in Y. lipolytica ( Cao et al, 2016 , 2017 ; Yang et al, 2016 ). Although truncated versions of Hmgp have been used in S. cerevisiae , studies indicate that the non-truncated version is superior for terpenoid production in Y. lipolytica ( Cao et al, 2016 ; Kildegaard et al, 2017 ; Bröker et al, 2018 ; Han et al, 2018 ). For example, the production of β-carotene was enhanced to a greater degree by HMG compared to tHMG both when solely expressed or in combination with the geranylgeranyl diphosphate synthase ( GGPPS ) ( Kildegaard et al, 2017 ).…”

Section: Resultsmentioning

confidence: 99%

Yarrowia lipolytica Strains Engineered for the Production of Terpenoids

Arnesen

Kildegaard

Pastor

et al. 2020

Front. Bioeng. Biotechnol.

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Terpenoids are a diverse group of over 55,000 compounds with potential applications as advanced fuels, bulk and fine chemicals, pharmaceutical ingredients, agricultural chemicals, etc. To facilitate their bio-based production, there is a need for plug-andplay hosts, capable of high-level production of different terpenoids. Here we engineer Yarrowia lipolytica platform strains for the overproduction of mono-, sesqui-, di-, tri-, and tetraterpenoids. The monoterpene platform strain was evaluated by expressing Perilla frutescens limonene synthase, which resulted in limonene titer of 35.9 mg/L and was 100-fold higher than when the same enzyme was expressed in the strain without mevalonate pathway improvement. Expression of Callitropsis nootkatensis valencene synthase in the sesquiterpene platform strain resulted in 113.9 mg/L valencene, an 8.4-fold increase over the control strain. Platform strains for production of squalene, complex triterpenes, or diterpenes and carotenoids were also constructed and resulted in the production of 402.4 mg/L squalene, 22 mg/L 2,3-oxidosqualene, or 164 mg/L β-carotene, respectively. The presented terpenoid platform strains can facilitate the evaluation of terpenoid biosynthetic pathways and are a convenient starting point for constructing efficient cell factories for the production of various terpenoids. The platform strains and exemplary terpenoid strains can be obtained from Euroscarf.

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High-level recombinant production of squalene using selectedSaccharomyces cerevisiaestrains

Cited by 43 publications

References 43 publications

Engineering Strategies in Microorganisms for the Enhanced Production of Squalene: Advances, Challenges and Opportunities

Engineering Strategies in Microorganisms for the Enhanced Production of Squalene: Advances, Challenges and Opportunities

Optimizing mevalonate pathway for squalene production inYarrowia lipolytica

Yarrowia lipolytica Strains Engineered for the Production of Terpenoids

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