scholarly journals Optimizing oleaginous yeast cell factories for flavonoids and hydroxylated flavonoids biosynthesis

2019 ◽  
Author(s):  
Yongkun Lv ◽  
Mattheos Koffas ◽  
Jingwen Zhou ◽  
Peng Xu
Keyword(s):  
Oleaginous Yeast ◽  
Heterologous Production ◽  
Modular Construction ◽  
Metabolic Potential ◽  
Cell Factories ◽  
Plant Extraction ◽  
Health Promoting ◽  
Product Manufacturing ◽  
Hydrophobic Lipid ◽  
Flavonoids Biosynthesis

AbstractPlants possess myriads of secondary metabolites with a broad spectrum of health-promoting benefits. Up to date, plant extraction is still the primary route to produce high-value natural products, which inherently suffers from economics and scalability issues. Heterologous production in microbial host is considered as a feasible approach to overcoming these limitations. Flavonoid and its hydroxylated derivatives represent a diversified family of bioactive compounds, most prominently known as antioxidant and anti-aging agents. Oleaginous yeast is rich in hydrophobic lipid bodies and spatially-organized organelles, which provides the ideal environment for the regioselectivity and stereoselectivity of many plant-specific enzymes. In this report, we validated thatY. lipolyticais a superior platform for heterologous production of high-value flavonoids and hydroxylated flavonoids. By modular construction and characterization, we determined the rate-limiting steps for efficient flavonoids biosynthesis inY. lipolytica. We evaluated various precursor pathways and unleashed the metabolic potential ofY. lipolyticato produce flavonoids, including the supply of acetyl-CoA, malonyl-CoA and chorismate. Coupled with the optimized chalcone synthase module and the hydroxylation module, our engineered strain produced 252.4 mg/L naringenin, 134.2 mg/L eriodictyol and 110.5 mg/L taxifolin from glucose. Collectively, these findings demonstrate our ability to harness oleaginous yeast as microbial workhorse to expand nature’s biosynthetic potential, enabling us to bridge the gap between drug discovery and natural product manufacturing.

scholarly journals Optimizing Oleaginous Yeast Cell Factories for Flavonoids and Hydroxylated Flavonoids Biosynthesis

2019 ◽  
Vol 8 (11) ◽  
pp. 2514-2523 ◽  
Author(s):  
Yongkun Lv ◽  
Monireh Marsafari ◽  
Mattheos Koffas ◽  
Jingwen Zhou ◽  
Peng Xu
Keyword(s):  
Yeast Cell ◽  
Oleaginous Yeast ◽  
Cell Factories ◽  
Flavonoids Biosynthesis

Heterologous production of pentane in the oleaginous yeast Yarrowia lipolytica

2013 ◽  
Vol 165 (3-4) ◽  
pp. 184-194 ◽  
Author(s):  
John Blazeck ◽  
Leqian Liu ◽  
Rebecca Knight ◽  
Hal S. Alper
Keyword(s):  
Yarrowia Lipolytica ◽  
Oleaginous Yeast ◽  
Heterologous Production ◽  
Yeast Yarrowia Lipolytica

scholarly journals Nonribosomal peptides in fungal cell factories: from genome mining to optimized heterologous production

2019 ◽  
Vol 37 (8) ◽  
pp. 107449 ◽  
Author(s):  
Antoine Vassaux ◽  
Loïc Meunier ◽  
Micheline Vandenbol ◽  
Denis Baurain ◽  
Patrick Fickers ◽  
...  
Keyword(s):  
Genome Mining ◽  
Nonribosomal Peptides ◽  
Heterologous Production ◽  
Fungal Cell ◽  
Cell Factories

scholarly journals Recent Advances on Feasible Strategies for Monoterpenoid Production in Saccharomyces cerevisiae

2020 ◽  
Vol 8 ◽  
Author(s):  
Qiyu Gao ◽  
Luan Wang ◽  
Maosen Zhang ◽  
Yongjun Wei ◽  
Wei Lin
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plant Biomass ◽  
Extraction Methods ◽  
Active Components ◽  
Plant Essential Oils ◽  
Microbial Cell Factories ◽  
Cell Factories ◽  
Plant Extraction ◽  
Low Efficiency ◽  
Metabolic Activities

Terpenoids are a large diverse group of natural products which play important roles in plant metabolic activities. Monoterpenoids are the main components of plant essential oils and the active components of some traditional Chinese medicinal herbs. Some monoterpenoids are widely used in medicine, cosmetics and other industries, and they are mainly obtained by plant biomass extraction methods. These plant extraction methods have some problems, such as low efficiency, unstable quality, and high cost. Moreover, the monoterpenoid production from plant cannot satisfy the growing monoterpenoids demand. The development of metabolic engineering, protein engineering and synthetic biology provides an opportunity to produce large amounts of monoterpenoids eco-friendly using microbial cell factories. This mini-review covers current monoterpenoids production using Saccharomyces cerevisiae. The monoterpenoids biosynthetic pathways, engineering of key monoterpenoids biosynthetic enzymes, and current monoterpenoids production using S. cerevisiae were summarized. In the future, metabolically engineered S. cerevisiae may provide one possible green and sustainable strategy for monoterpenoids supply.

scholarly journals Enhancement of Astaxanthin Biosynthesis in Oleaginous Yeast Yarrowia lipolytica via Microalgal Pathway

2019 ◽  
Vol 7 (10) ◽  
pp. 472 ◽  
Author(s):  
Larissa Ribeiro Ramos Tramontin ◽  
Kanchana Rueksomtawin Kildegaard ◽  
Suresh Sudarsan ◽  
Irina Borodina
Keyword(s):  
Yarrowia Lipolytica ◽  
Oleaginous Yeast ◽  
Small Scale ◽  
Cell Factory ◽  
Standard Equipment ◽  
Cell Factories ◽  
Dry Cell Weight ◽  
Astaxanthin Production ◽  
Yeast Yarrowia Lipolytica ◽  
Β Carotene

Astaxanthin is a high-value red pigment and antioxidant used by pharmaceutical, cosmetics, and food industries. The astaxanthin produced chemically is costly and is not approved for human consumption due to the presence of by-products. The astaxanthin production by natural microalgae requires large open areas and specialized equipment, the process takes a long time, and results in low titers. Recombinant microbial cell factories can be engineered to produce astaxanthin by fermentation in standard equipment. In this work, an oleaginous yeast Yarrowia lipolytica was engineered to produce astaxanthin at high titers in submerged fermentation. First, a platform strain was created with an optimised pathway towards β-carotene. The platform strain produced 331 ± 66 mg/L of β-carotene in small-scale cultivation, with the cellular content of 2.25% of dry cell weight. Next, the genes encoding β-ketolase and β-hydroxylase of bacterial (Paracoccus sp. and Pantoea ananatis) and algal (Haematococcus pluvialis) origins were introduced into the platform strain in different copy numbers. The resulting strains were screened for astaxanthin production, and the best strain, containing algal β-ketolase and β-hydroxylase, resulted in astaxanthin titer of 44 ± 1 mg/L. The same strain was cultivated in controlled bioreactors, and a titer of 285 ± 19 mg/L of astaxanthin was obtained after seven days of fermentation on complex medium with glucose. Our study shows the potential of Y. lipolytica as the cell factory for astaxanthin production.

scholarly journals Genome-scale metabolic reconstruction of the stress-tolerant hybrid yeast Zygosaccharomyces parabailii

2018 ◽  
Author(s):  
Marzia Di Filippo ◽  
Raúl A. Ortiz-Merino ◽  
Chiara Damiani ◽  
Gianni Frascotti ◽  
Danilo Porro ◽  
...  
Keyword(s):  
Laboratory Data ◽  
Metabolic Model ◽  
Cellular Systems ◽  
Metabolic Reconstruction ◽  
Genome Sequences ◽  
Metabolic Potential ◽  
Cell Factories ◽  
Metabolic Models ◽  
Constraint Based Modeling ◽  
Genome Scale

Genome-scale metabolic models are powerful tools to understand and engineer cellular systems facilitating their use as cell factories. This is especially true for microorganisms with known genome sequences from which nearly complete sets of enzymes and metabolic pathways are determined, or can be inferred. Yeasts are highly diverse eukaryotes whose metabolic traits have long been exploited in industry, and although many of their genome sequences are available, few genome-scale metabolic models have so far been produced. For the first time, we reconstructed the genome-scale metabolic model of the hybrid yeast Zygosaccharomyces parabailii, which is a member of the Z. bailii sensu lato clade notorious for stress-tolerance and therefore relevant to industry. The model comprises 3096 reactions, 2091 metabolites, and 2413 genes. Our own laboratory data were then used to establish a biomass synthesis reaction, and constrain the extracellular environment. Through constraint-based modeling, our model reproduces the co-consumption and catabolism of acetate and glucose posing it as a promising platform for understanding and exploiting the metabolic potential of Z. parabailii.

scholarly journals Biovalorisation of crude glycerol and xylose into xylitol by oleaginous yeast Yarrowia lipolytica

2020 ◽  
Author(s):  
Ashish Prabhu ◽  
Dominic J Thomas ◽  
Rodrigo Ledesma- Amaro ◽  
Gary A Leeke ◽  
Angel Medina Vaya ◽  
...  
Keyword(s):  
Yarrowia Lipolytica ◽  
Crude Glycerol ◽  
Oleaginous Yeast ◽  
Scale Up ◽  
Microbial Cell ◽  
Department Of Energy ◽  
Cell Factory ◽  
Potential Cost ◽  
Cell Factories ◽  
Pharmaceutical Industries

Abstract Background: Xylitol is a commercially important chemical with multiple applications in the food and pharmaceutical industries. According to the US Department of Energy, xylitol is one of the top twelve platform chemicals that can be produced from biomass. The chemical method for xylitol synthesis is however expensive and energy intensive. In contrast, the biological route using microbial cell factories offers a potential cost-effective alternative process. The bioprocess occurs under ambient conditions and makes use of biocatalysts and biomass which can be sourced from renewable carbon originating from a variety of cheap waste feedstocks. Result: In this study, biotransformation of xylose to xylitol was investigated using Yarrowia lipolytica an oleaginous yeast which was firstly grown on a glycerol/glucose for screening of co-substrate, followed by media optimisation in shake flask, scale up in bioreactor and downstream processing of xylitol. A two-step medium optimization was employed using central composite design and artificial neural network coupled with genetic algorithm. The yeast amassed a concentration of 53.2 g/L xylitol using pure glycerol (PG) and xylose with a bioconversion yield of 0.97 g/g. Similar results were obtained when PG was substituted with crude glycerol (CG) from the biodiesel industry (titer: 50.5 g/L; yield: 0.92 g/g). Even when xylose from sugarcane bagasse hydrolysate was used as opposed to pure xylose, a xylitol yield of 0.54 g/g was achieved. Xylitol was successfully crystallized from PG/xylose and CG /xylose fermentation broths with a recovery of 39.5 and 35.3%, respectively. Conclusion: To the best of the author’s knowledge, this study demonstrates for the first time the potential of using Y. lipolytica as a microbial cell factory for xylitol synthesis from inexpensive feedstocks. The results obtained are competitive with other xylitol producing organisms.

scholarly journals Engineered gray mold resistance, antioxidant capacity, and pigmentation in betalain-producing crops and ornamentals

2017 ◽  
Vol 114 (34) ◽  
pp. 9062-9067 ◽  
Author(s):  
Guy Polturak ◽  
Noam Grossman ◽  
David Vela-Corcia ◽  
Yonghui Dong ◽  
Adi Nudel ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Crop Protection ◽  
Gray Mold ◽  
Food Crops ◽  
Heterologous Production ◽  
Tobacco Plants ◽  
Agricultural Produce ◽  
Health Promoting ◽  
Mold Resistance ◽  
Combinatorial Expression

Betalains are tyrosine-derived red-violet and yellow plant pigments known for their antioxidant activity, health-promoting properties, and wide use as food colorants and dietary supplements. By coexpressing three genes of the recently elucidated betalain biosynthetic pathway, we demonstrate the heterologous production of these pigments in a variety of plants, including three major food crops: tomato, potato, and eggplant, and the economically important ornamental petunia. Combinatorial expression of betalain-related genes also allowed the engineering of tobacco plants and cell cultures to produce a palette of unique colors. Furthermore, betalain-producing tobacco plants exhibited significantly increased resistance toward gray mold (Botrytis cinerea), a pathogen responsible for major losses in agricultural produce. Heterologous production of betalains is thus anticipated to enable biofortification of essential foods, development of new ornamental varieties, and innovative sources for commercial betalain production, as well as utilization of these pigments in crop protection.

scholarly journals Bioactive Phenolic Compounds from Primula veris L.: Influence of the Extraction Conditions and Purification

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 997
Author(s):  
Maria Tarapatskyy ◽  
Aleksandra Gumienna ◽  
Patrycja Sowa ◽  
Ireneusz Kapusta ◽  
Czesław Puchalski
Keyword(s):  
Selective Extraction ◽  
Bioactive Substances ◽  
Plant Extraction ◽  
Primula Veris ◽  
Health Promoting ◽  
Phase Liquid ◽  
Efficient Extraction ◽  
Ethanol Extracts ◽  
The Individual

Our experiments may help to answer the question of whether cowslip (Primula veris L.) is a rich source of bioactive substances that can be obtained by efficient extraction with potential use as a food additive. A hypothesis assumed that the type of solvent used for plant extraction and the individual morphological parts of Primula veris L. used for the preparation of herbal extracts will have key impacts on the efficiency of the extraction of bioactive compounds, and thus, the health-promoting quality of plant concentrates produced. Most analysis of such polyphenolic compound contents in extracts from Primula veris L. has been performed by using chromatography methods such as ultra-performance reverse-phase liquid chromatography (UPLC−PDA−MS/MS). Experiments demonstrated that the most effective extraction agent for fresh study material was water at 100 °C, whereas for dried material it was 70% ethanol. The richest sources of polyphenolic compounds were found in cowslip primrose flowers and leaves. The aqueous and ethanol extracts from Primula veris L. were characterized by a quantitatively rich profile of polyphenolic substances, and a high antioxidative potential. Selective extraction with the use of mild conditions and neutral solvents is the first step to obtaining preparations from cowslip primrose with a high content of bioactive substances.

scholarly journals Increased accumulation of squalene in engineered Yarrowia lipolytica through deletion of PEX10 and URE2

2021 ◽  
Author(s):  
Liu-Jing Wei ◽  
Xuan Cao ◽  
Jing-Jing Liu ◽  
Suryang Kwak ◽  
Yong-Su Jin ◽  
...  
Keyword(s):  
Yarrowia Lipolytica ◽  
Catabolite Repression ◽  
Oleaginous Yeast ◽  
Mevalonate Pathway ◽  
Culture Media ◽  
Yeast Cells ◽  
Nitrogen Catabolite Repression ◽  
Peroxisomal Membrane ◽  
Cell Factories ◽  
Pharmaceutical Industries

Squalene is a triterpenoid serving as an ingredient of various products in the food, cosmetic, pharmaceutical industries. The oleaginous yeast Yarrowia lipolytica offers enormous potential as a microbial chassis for the production of terpenoids, such as carotenoid, limonene, linalool, and farnesene as the yeast provides ample storage space for hydrophobic products. Here we present a metabolic design that allows the enhanced accumulation of squalene in Y. lipolytica . First, we improved squalene accumulation in Y. lipolytica by overexpressing the genes (ERG, HMG) coding for the mevalonate pathway enzymes. Second, we increased the production of lipid where squalene is accumulated by overexpressing DGA1 encoding for diacylglycerol acyltransferase and deleting PEX10 for peroxisomal membrane E3 ubiquitin ligase. Third, we deleted URE2 coding for a transcriptional regulator in charge of nitrogen catabolite repression (NCR) to induce lipid accumulation regardless of carbon to nitrogen ratio in culture media. The resulting engineered Y. lipolytica exhibited a 115-fold higher squalene content (22.0 mg/g DCW) than a parental strain. These results suggest that the biological function of Ure2p in Y. lipolytica is similar to that in S. cerevisiae , and its deletion can be utilized to enhance the production of hydrophobic target products in oleaginous yeast strains. IMPORTANCE This study demonstrated a novel strategy for increasing squalene production in Y. lipolytica . URE2, a bifunctional protein that is involved in both nitrogen catabolite repression and oxidative stress response, was identified and demonstrated correlation to squalene production. The data suggest that double deletion of PEX10 and URE2 can serve as a positive synergistic effect to help yeast cells on boosting squalene production. This discovery can be combined with other strategies to engineer cell factories to efficiently produce terpenoid in the future.

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