Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA

Pereira, H.M.; Azevedo, Flávio; Domingues, Lucı́lia; Johansson, Börje

doi:10.1016/j.csbj.2022.01.020

Cited by 11 publications

(6 citation statements)

References 51 publications

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“…The yield in PpHP8 (harboring Acc1 Yl ) was around 1.25fold higher than in PpHP7. These results are consistent with recent studies where the heterologous expression of ACC1 from Y. lipolytica yielded a higher accumulation of malonyl-CoA than the overexpression of the endogenous S. cerevisiae ACC1 (Pereira et al, 2022).…”

Section: Expression Of the De-regulated Acc1 S1132a Mutant Does Not I...supporting

confidence: 93%

Combining Metabolic Engineering and Multiplexed Screening Methods for 3-Hydroxypropionic Acid Production in Pichia pastoris

Fina

Heux

Albiol

et al. 2022

Front. Bioeng. Biotechnol.

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Production of 3-hydroxypropionic acid (3-HP) in Pichia pastoris (syn. Komagataella phaffii) via the malonyl-CoA pathway has been recently demonstrated using glycerol as a carbon source, but the reported metrics were not commercially relevant. The flux through the heterologous pathway from malonyl-CoA to 3-HP was hypothesized as the main bottleneck. In the present study, different metabolic engineering approaches have been combined to improve the productivity of the original 3-HP producing strains. To do so, an additional copy of the gene encoding for the potential rate-limiting step of the pathway, i.e., the C-terminal domain of the malonyl-CoA reductase, was introduced. In addition, a variant of the endogenous acetyl-CoA carboxylase (ACC1S1132A) was overexpressed with the aim to increase the delivery of malonyl-CoA. Furthermore, the genes encoding for the pyruvate decarboxylase, aldehyde dehydrogenase and acetyl-CoA synthase, respectively, were overexpressed to enhance conversion of pyruvate into cytosolic acetyl-CoA, and the main gene responsible for the production of the by-product D-arabitol was deleted. Three different screening conditions were used to classify the performance of the different strains: 24-deep-well plates batch cultures, small-scale cultures in falcon tubes using FeedBeads® (i.e., slow release of glycerol over time), and mini bioreactor batch cultures. The best two strains from the FeedBeads® screening, PpHP8 and PpHP18, were tested in bioreactor fed-batch cultures using a pre-fixed exponentially increasing feeding rate. The strain PpHP18 produced up to 37.05 g L−1 of 3-HP at 0.712 g L−1 h−1 with a final product yield on glycerol of 0.194 Cmol−1 in fed-batch cultures. Remarkably, PpHP18 did not rank among the 2-top producer strains in small scale batch cultivations in deep-well plates and mini bioreactors, highlighting the importance of multiplexed screening conditions for adequate assessment of metabolic engineering strategies. These results represent a 50% increase in the product yield and final concentration, as well as over 30% increase in volumetric productivity compared to the previously obtained metrics for P. pastoris. Overall, the combination of glycerol as carbon source and a metabolically engineered P. pastoris strain resulted in the highest 3-HP concentration and productivity reported so far in yeast.

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Section: Expression Of the De-regulated Acc1 S1132a Mutant Does Not I...supporting

confidence: 93%

Combining Metabolic Engineering and Multiplexed Screening Methods for 3-Hydroxypropionic Acid Production in Pichia pastoris

Fina

Heux

Albiol

et al. 2022

Front. Bioeng. Biotechnol.

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“…The different fat content might potentially affect the efficiency of BNC fermentation when utilizing extracts derived from these distinct yeast species. 33 The observed enhancement in BNC production could be attributed to potential variations in amino acid composition between the extracts from Y. lipolytica and S. cerevisiae, and the Y. lipolytica extract may be more conducive to BNC fermentation. However, the BNC production was only 48.2% of the control when using the Y. lipolytica extract (ground and lyticase + protease treated), yielding 1.16 g/L.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…cerevisiae yeast. The different fat content might potentially affect the efficiency of BNC fermentation when utilizing extracts derived from these distinct yeast species . The observed enhancement in BNC production could be attributed to potential variations in amino acid composition between the extracts from Y.…”

Section: Resultsmentioning

confidence: 99%

Production of High-Value-Added Bacterial Nanocellulose with Thinner Fiber Using Novel Yarrowia lipolytica Extract from Erythritol Industry Waste

Sun,

Tan,

Yan

et al. 2024

ACS Sustainable Chem. Eng.

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The rapid growth of the erythritol fermentation industry has prompted interest in the efficient utilization of its yeast cell residue. This study concentrates on extracting nitrogen sources from Yarrowia lipolytica waste to enhance microbial fermentation of bacterial nanocellulose (BNC), a valuable nanoscale biopolymer. Various treatments, including enzymatic, mechanical, and alkali treatments at different temperatures, were employed to improve extraction. The most effective treatments were alkali treatments at 50 and 90 °C, followed by lyticase and protease treatments. The highest production of soluble protein/amino acid contents reached 1.997 ± 0.018 and 2.328 ± 0.018 g/L, respectively. The 50 °C treatment was the most cost-effective based on thermal energy calculations and was selected for further analysis. Y. lipolytica yeast extract enhanced BNC production to 4.017 g/L, a 66.5% improvement over the commercial Saccharomyces cerevisiae yeast extract. Substituting half of the commercial yeast extract with the Y. lipolytica extract also increased BNC production by 22.6%. No significant differences were observed in the chemical composition, crystallinity, and thermal stabilities of BNC prepared from Y. lipolytica extract compared to those of commercial yeast extract. Transmission electron microscopy (TEM) revealed thinner fiber diameters in BNC prepared from Y. lipolytica extract, and atomic force microscopy (AFM) measurements showed a remarkable 189.1% increase in surface roughness.

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“…The study of fatty acid metabolism, especially in pt55_3, clarified the influence of grazing pressure on the biosynthesis, elongation, and degradation pathways. Within the fatty acid biosynthesis pathway ( Supplementary Figure 16 ), a significant down-regulation of ACACA (Acetyl-CoA carboxylase) implies a reduction in ATP-dependent carboxylation of Acetyl-CoA to Malonyl-CoA under grazing pressure ( Pereira et al., 2022 ). Enoyl ACP reductases, including FabI and its isomers (FabL, FabK, FabV, and InhA), play a crucial role in many microorganisms ( Rana et al., 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Multi-omic insights into the cellular response of Phaeodactylum tricornutum (Bacillariophyta) strains under grazing pressure

Liu,

Li,

Yang

et al. 2024

Front. Plant Sci.

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Background/AimsPhaeodactylum tricornutum, a model organism of diatoms, plays a crucial role in Earth’s primary productivity. Investigating its cellular response to grazing pressure is highly significant for the marine ecological environment. Furthermore, the integration of multi-omics approaches has enhanced the understanding of its response mechanism.MethodsTo assess the molecular and cellular responses of P.tricornutum to grazer presence, we conducted transcriptomic, proteomic, and metabolomic analyses, combined with phenotypic data from previous studies. Sequencing data were obtained by Illumina RNA sequencing, TMT Labeled Quantitative Proteomics and Non-targeted Metabolomics, and WGCNA analysis and statistical analysis were performed.ResultsAmong the differentially expressed genes, we observed complex expression patterns of the core genes involved in the phenotypic changes of P.tricornutum under grazing pressure across different strains and multi-omics datasets. These core genes primarily regulate the levels of various proteins and fatty acids, as well as the cellular response to diverse signals.ConclusionOur research reveals the association of multi-omics in four strains responses to grazing effects in P.tricornutum. Grazing pressure significantly impacted cell growth, fatty acid composition, stress response, and the core genes involved in phenotype transformation.

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Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA

Cited by 11 publications

References 51 publications

Combining Metabolic Engineering and Multiplexed Screening Methods for 3-Hydroxypropionic Acid Production in Pichia pastoris

Combining Metabolic Engineering and Multiplexed Screening Methods for 3-Hydroxypropionic Acid Production in Pichia pastoris

Production of High-Value-Added Bacterial Nanocellulose with Thinner Fiber Using Novel Yarrowia lipolytica Extract from Erythritol Industry Waste

Multi-omic insights into the cellular response of Phaeodactylum tricornutum (Bacillariophyta) strains under grazing pressure

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