Genetic and bioprocess engineering to improve squalene production in Yarrowia lipolytica

Wang

Critical Reviews in Biotechnology

et al. 2021

Terpenoids are a large family of natural products with diversified structures and functions that are widely used in the food, pharmaceutical, cosmetic, and agricultural fields. However, the traditional methods of terpenoids production such as plant extraction and chemical synthesis are inefficient due to the complex processes, high energy consumption, and low yields. With progress in metabolic engineering and synthetic biology, microbial cell factories provide an interesting alternative for the sustainable production of terpenoids. The non-conventional yeast, Yarrowia lipolytica, is a promising host for terpenoid biosynthesis due to its inherent mevalonate pathway, high fluxes of acetyl-CoA and NADPH, and the naturally hydrophobic microenvironment. In this review, we highlight progress in the engineering of Y. lipolytica as terpenoid biomanufacturing factories, describing the different terpenoid biosynthetic pathways and summarizing various metabolic engineering strategies, including progress in genetic manipulation, dynamic regulation, organelle engineering, and terpene synthase variants.

Section: Engineering the Cofactor Supplymentioning

confidence: 99%

Metabolic engineering of Yarrowia lipolytica for terpenoids production: advances and perspectives

Wang

Critical Reviews in Biotechnology

et al. 2021

Biotech & Bioengineering

“…In case of lipogenesis, this is also since the lipid metabolism has proven to be quite complex in case of Y. lipolytica and it indicates the need of incorporating more data into genome‐scale models as discussed before. Moreover, multiple approaches can be used to overcome this issue amongst which bioprocess engineering (H. Liu et al, 2020), use of mixed substrates to optimize metabolism (N. Liu et al, 2020), and cellular engineering (Soong et al, 2019) can be named, discussion on which is out of the scope of this review. However, implementing such approaches along with existing metabolic engineering methods can potentially enhance cell performance.…”

Section: Systems Metabolic Engineering Of Y Lipolyticamentioning

confidence: 99%

Systems‐level approaches for understanding and engineering of the oleaginous cell factory Yarrowia lipolytica

Poorinmohammad

Kerkhoven

2021

Concerns about climate change and the search for renewable energy sources together with the goal of attaining sustainable product manufacturing have boosted the use of microbial platforms to produce fuels and high‐value chemicals. In this regard, Yarrowia lipolytica has been known as a promising yeast with potentials in diverse array of biotechnological applications such as being a host for different oleochemicals, organic acid, and recombinant protein production. Having a rapidly increasing number of molecular and genetic tools available, Y. lipolytica has been well studied amongst oleaginous yeasts and metabolic engineering has been used to explore its potentials. More recently, with the advancement in systems biotechnology and the implementation of mathematical modeling and high throughput omics data‐driven approaches, in‐depth understanding of cellular mechanisms of cell factories have been made possible resulting in enhanced rational strain design. In case of Y. lipolytica, these systems‐level studies and the related cutting‐edge technologies have recently been initiated which is expected to result in enabling the biotechnology sector to rationally engineer Y. lipolytica‐based cell factories with favorable production metrics. In this regard, here, we highlight the current status of systems metabolic engineering research and assess the potential of this yeast for future cell factory design development.

“…Different from Saccharomyces cerevisiae , Y. lipolytica lacks Crabtree effects and does not produce ethanol when carbohydrates are oversupplied in the media. 19 Y. lipolytica has been reported to grow on a wide range of inexpensive raw materials, 20 including glucose, 21 glycerol, 22 xylose, 20 , 23 volatile fatty acids, 24 − 26 alcohols, 24 and wax alkanes. 27 …”

Section: Introductionmentioning

confidence: 99%

A Golden-Gate Based Cloning Toolkit to Build Violacein Pathway Libraries in Yarrowia lipolytica

et al. 2021

Self Cite

Violacein is a naturally occurring anticancer therapeutic compound with deep purple color. In this work, we harnessed the modular and combinatorial feature of a Golden Gate assembly method to construct a library of violacein producing strains in the oleaginous yeast Yarrowia lipolytica, where each gene in the violacein pathway was controlled by three different promoters with varying transcriptional strength. After optimizing the linker sequence and the Golden Gate reaction, we achieved high transformation efficiency and obtained a panel of representative Y. lipolytica recombinant strains. By evaluating the gene expression profile of 21 yeast strains, we obtained three colorful compounds in the violacein pathway: green (proviolacein), purple (violacein), and pink (deoxyviolacein). Our results indicated that strong expression of VioB, VioC, and VioD favors violacein production with minimal byproduct deoxyvioalcein in Y. lipolytica, and high deoxyviolacein production was found strongly associated with the weak expression of VioD. By further optimizing the carbon to nitrogen ratio and cultivation pH, the maximum violacein reached 70.04 mg/L with 5.28 mg/L of deoxyviolacein in shake flasks. Taken together, the development of Golden Gate cloning protocols to build combinatorial pathway libraries, and the optimization of culture conditions set a new stage for accessing the violacein pathway intermediates and engineering violacein production in Y. lipolytica. This work further expands the toolbox to engineering Y. lipolytica as an industrially relevant host for plant or marine natural product biosynthesis.