Production of β-carotene by expressing a heterologous multifunctional carotene synthase in Yarrowia lipolytica

Gao, Shuliang; Tong, Yongliang; Zhu, Li; Ge, Mei; Jiang, Yu; Chen, Daijie; Yang, Sheng

doi:10.1007/s10529-017-2318-1

Cited by 34 publications

(17 citation statements)

References 20 publications

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“…In two of the current isolates (St8 and St9), β-carotene production was found to be higher per unit of biomass (15- and 8-fold, respectively) as compared to its production found in earlier thraustochytrids studies [17]. Additionally, recently developed transgenic strain produced about 30% more pigment as compared to the isolate St8 [56]. These findings indicate a relatively high and naturally occurring ratio of pigment per unit of biomass, which could point towards production potential if culture conditions are further optimized [57].…”

Section: Discussionmentioning

confidence: 94%

Isolation, Characterization and Biotechnological Potentials of Thraustochytrids from Icelandic Waters

et al. 2019

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The following study reports on the first thraustochytrid isolates identified from Iceland. They were collected from three different locations off the northern coast of the country (Location A, Skagaströnd; Location B, Hveravík; and Location C, Eyjafjörður). Using 18S rDNA sequence analysis, isolates from Locations A and B were identified within the Thraustochytrium kinnei species while other isolates within the Sicyoidochytrium minutum species when compared to other known strains. Cells isolated from Locations A ( 2 . 10 ± 0 . 70 g/L) and B ( 1 . 54 ± 0 . 17 g/L) produced more biomass than the ones isolated from Location C ( 0 . 43 ± 0 . 02 g/L). This study offers the first-time examination of the utility of byproducts from fisheries as a nitrogen source in media formulation for thraustochytrids. Experiments showed that isolates produced more biomass (per unit of substrate) when cultured on nitrogen of marine ( 2 . 55 ± 0 . 74 g/L) as compared to of commercial origin ( 1 . 06 ± 0 . 57 g/L). Glycerol ( 2 . 43 ± 0 . 56 g/L) was a better carbon source than glucose ( 1 . 84 ± 0 . 57 g/L) in growth studies. Fatty acid (FA) profiles showed that the isolates from Location C (S. minutum) had low ratios of monounsaturated ( 4 . 21 ± 2 . 96 % ) and omega-6 ( 0 . 68 ± 0 . 59 % ) FAs. However, the isolates also had high ratios of docosahexaenoic acid (DHA; 35 . 65 ± 1 . 73 % ) and total omega-3 FAs ( 40 . 39 ± 2 . 39 % ), indicating that they could serve as a source of marine oils for human consumption and in aquaculture feeds. The T. kinnei isolates from Location A could be used in biodiesel production due to their high ratios of monounsaturated ( 18 . 38 ± 6 . 27 % ) long chain ( 57 . 43 ± 8 . 27 % ) FAs.

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Section: Discussionmentioning

confidence: 94%

Isolation, Characterization and Biotechnological Potentials of Thraustochytrids from Icelandic Waters

et al. 2019

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“…Similarly, the natural tridomain fusion (CrtIBY) from Schizotrium sp. was expressed recently in Yarrowia lipolytica (Gao et al, 2017) but the β-carotene production yields were considerably lower than those obtained with the yeast-expressed X. dendrorhous configuration (Verwaal et al, 2007, Xie et al, 2015c). Although these heterologous systems, based on X. dendrorhous enzymes, are the most productive to date, accumulation of β-carotene precursors in the pathway, such as phytoene, was evidenced.…”

Section: Introductionmentioning

confidence: 92%

Enzyme-fusion strategies for redirecting and improving carotenoid synthesis in S. cerevisiae

Rabeharindranto

Castaño-Cerezo

Lautier

et al. 2019

Metabolic Engineering Communications

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Spatial clustering of enzymes has proven an elegant approach to optimize metabolite transfer between enzymes in synthetic metabolic pathways. Among the multiple methods used to promote colocalisation, enzyme fusion is probably the simplest. Inspired by natural systems, we have explored the metabolic consequences of spatial reorganizations of the catalytic domains of Xanthophyllomyces dendrorhous carotenoid enzymes produced in Saccharomyces cerevisiae. Synthetic genes encoding bidomain enzymes composed of CrtI and CrtB domains from the natural CrtYB fusion were connected in the two possible orientations, using natural and synthetic linkers. A tridomain enzyme (CrtB, CrtI, CrtY) harboring the full β-carotene producing pathway was also constructed. Our results demonstrate that domain order and linker properties considerably impact both the expression and/or stability of the constructed proteins and the functionality of the catalytic domains, all concurring to either diminish or boost specific enzymatic steps of the metabolic pathway. Remarkably, the yield of β-carotene production doubled with the tridomain fusion while precursor accumulation decreased, leading to an improvement of the pathway efficiency, when compared to the natural system. Our data strengthen the idea that fusion of enzymatic domains is an appropriate technique not only to achieve spatial confinement and enhance the metabolic flux but also to produce molecules not easily attainable with natural enzymatic configurations, even with membrane bound enzymes.

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“…This requires high flux through acetyl-CoA, making it a good production host for molecules that likewise require acetyl-CoA. There are numerous examples of metabolic engineering efforts to make economically useful amounts of products, including TAGs, omega-3 rich TAGs, fatty acid derivatives, carotenoids, and organic acids ( Yuzbashev et al, 2010 ; Xue et al, 2013 ; Blazeck et al, 2014 ; Qiao et al, 2015 ; Ryu et al, 2015 ; Wang et al, 2016 ; Xu et al, 2016 ; Gao et al, 2017 ; Kildegaard et al, 2017 ; Sabra et al, 2017 ; Schwartz et al, 2017a ; Xie et al, 2017 ; Sagnak et al, 2018 ). Several recent reviews have thoroughly covered metabolic engineering efforts, as well as advances in the genetic engineering toolkit that enables such work ( Blazeck and Alper, 2013 ; Hussain et al, 2016 ; Adrio, 2017 ; Yan et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Alternative Substrate Metabolism in Yarrowia lipolytica

et al. 2018

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Recent advances in genetic engineering capabilities have enabled the development of oleochemical producing strains of Yarrowia lipolytica. Much of the metabolic engineering effort has focused on pathway engineering of the product using glucose as the feedstock; however, alternative substrates, including various other hexose and pentose sugars, glycerol, lipids, acetate, and less-refined carbon feedstocks, have not received the same attention. In this review, we discuss recent work leading to better utilization of alternative substrates. This review aims to provide a comprehensive understanding of the current state of knowledge for alternative substrate utilization, suggest potential pathways identified through homology in the absence of prior characterization, discuss recent work that either identifies, endogenous or cryptic metabolism, and describe metabolic engineering to improve alternative substrate utilization. Finally, we describe the critical questions and challenges that remain for engineering Y. lipolytica for better alternative substrate utilization.

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Production of β-carotene by expressing a heterologous multifunctional carotene synthase in Yarrowia lipolytica

Cited by 34 publications

References 20 publications

Isolation, Characterization and Biotechnological Potentials of Thraustochytrids from Icelandic Waters

Isolation, Characterization and Biotechnological Potentials of Thraustochytrids from Icelandic Waters

Enzyme-fusion strategies for redirecting and improving carotenoid synthesis in S. cerevisiae

Alternative Substrate Metabolism in Yarrowia lipolytica

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