Enhancing isoprenoid synthesis in Yarrowia lipolytica by expressing the isopentenol utilization pathway and modulating intracellular hydrophobicity

Luo, Zhengshan; Liu, Nian; Lazar, Zbigniew; Chatzivasileiou, Alkiviadis Orfefs; Ward, Valerie C.A.; Chen, Jian; Zhou, Jingwen; Stephanopoulos, Gregory

doi:10.1016/j.ymben.2020.07.010

Cited by 82 publications

(105 citation statements)

References 36 publications

Supporting

Mentioning

102

Contrasting

Order By: Relevance

“…During the last year, the productivity of these engineered biofactories has been improved significantly, mainly by engineering gene expression, improving lycopene storage capacity, optimizing fermentation processes, and combinatorial pathway engineering (Li et al, 2020). The highest lycopene titer achieved thus far is 4.2 g per L culture medium in the yeast Y. lipolytica, which was achieved by a combination of pathway engineering and improved fermentation strategies (Luo et al, 2020). The yield in the BYL system was significantly lower (21 µg/mL) but this is, to our knowledge, the first time lycopene has been produced in a cell-free expression system and thus provides a versatile platform for the production optimization of carotenoids and other high-value isoprenoids.…”

Section: Discussionmentioning

confidence: 99%

Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites

et al. 2022

View full text Add to dashboard Cite

Cell-free expression systems enable the production of proteins and metabolites within a few hours or days. Removing the cellular context while maintaining the protein biosynthesis apparatus provides an open system that allows metabolic pathways to be installed and optimized by expressing different numbers and combinations of enzymes. This facilitates the synthesis of secondary metabolites that are difficult to produce in cell-based systems because they are toxic to the host cell or immediately converted into downstream products. Recently, we developed a cell-free lysate derived from tobacco BY-2 cell suspension cultures for the production of recombinant proteins. This system is remarkably productive, achieving yields of up to 3 mg/mL in a one-pot in vitro transcription–translation reaction and contains highly active energy and cofactor regeneration pathways. Here, we demonstrate for the first time that the BY-2 cell-free lysate also allows the efficient production of several classes of secondary metabolites. As case studies, we synthesized lycopene, indigoidine, betanin, and betaxanthins, which are useful in the food, cosmetic, textile, and pharmaceutical industries. Production was achieved by the co-expression of up to three metabolic enzymes. For all four products, we achieved medium to high yields. However, the yield of betanin (555 μg/mL) was outstanding, exceeding the level reported in yeast cells by a factor of more than 30. Our results show that the BY-2 cell-free lysate is suitable not only for the verification and optimization of metabolic pathways, but also for the efficient production of small to medium quantities of secondary metabolites.

show abstract

Section: Discussionmentioning

confidence: 99%

Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites

et al. 2022

View full text Add to dashboard Cite

show abstract

“…Yang et al [43] reported that IDI overexpression increased the titer of a-farnesene to 57.08 mg/L, nearly 2.67-fold higher than that produced by the wild-type strain. In addition to overexpressing PvIDI from Pseudescherichia vulneris, Luo et al [63] introduced a two-step isoprenol utilization pathway (AtIPK-ScCHK) in Y. lipolytica to enhance the supply of IPP and DMAPP. This chimeric AtIPK-ScCHK pathway elevated the total levels of IPP and DMAPP by 15.7-fold compared to the MVA pathway alone, which further improved the lycopene content to approximately 120 mg/g DCW.…”

Section: Overexpressing Mva Pathway Genesmentioning

confidence: 99%

“…Lipid synthesis through an ex novo pathway can be activated by feeding Y. lipolytica with exogenous fatty acids. Luo et al [63] enhanced lipid synthesis separately by overexpressing both ACC1 and DGA1 or supplementing palmitic acid and compared the effects of these two tactics on the yield of lycopene. The addition of palmitic acid to strains resulted in a lycopene triter of 594.6 mg/L, much higher compared with the controls (less than 400 mg/L).…”

Section: Modulating Hydrophobicity For Terpenoid Accumulationmentioning

confidence: 99%

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

Zhang

Wang

Zhang

et al. 2021

Critical Reviews in Biotechnology

View full text Add to dashboard Cite

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.

show abstract

“…Very recently, some reports using the TMP together with an engineered MVA pathway described the production of lycopene, geranylgeraniol, and α-farnesene in various yeasts ( Scheme 10 ). Luo et al [ 33 ] took advantage of the oleaginous nature of the yeast Yarrowia lipolytica to produce lycopene with the rationale that the lipid body of this yeast species could store more hydrophobic lycopene than the membranes. It was demonstrated that adding the genes coding for choline kinase and IPK results in enhanced intracellular production of IPP/DMAPP upon IOH addition.…”

Section: Two Enzyme Access To Dmapp/ippmentioning

confidence: 99%

The Terpene Mini-Path, a New Promising Alternative for Terpenoids Bio-Production

Couillaud

Leydet

Duquesne

et al. 2021

Genes

View full text Add to dashboard Cite

Terpenoids constitute the largest class of natural compounds and are extremely valuable from an economic point of view due to their extended physicochemical properties and biological activities. Due to recent environmental concerns, terpene extraction from natural sources is no longer considered as a viable option, and neither is the chemical synthesis to access such chemicals due to their sophisticated structural characteristics. An alternative to produce terpenoids is the use of biotechnological tools involving, for example, the construction of enzymatic cascades (cell-free synthesis) or a microbial bio-production thanks to metabolic engineering techniques. Despite outstanding successes, these approaches have been hampered by the length of the two natural biosynthetic routes (the mevalonate and the methyl erythritol phosphate pathways), leading to dimethylallyl diphosphate (DMAPP) and isopentenyl diphosphate (IPP), the two common universal precursors of all terpenoids. Recently, we, and others, developed what we called the terpene mini-path, a robust two enzyme access to DMAPP and IPP starting from their corresponding two alcohols, dimethylallyl alcohol and isopentenol. The aim here is to present the potential of this artificial bio-access to terpenoids, either in vitro or in vivo, through a review of the publications appearing since 2016 on this very new and fascinating field of investigation.

show abstract

Enhancing isoprenoid synthesis in Yarrowia lipolytica by expressing the isopentenol utilization pathway and modulating intracellular hydrophobicity

Cited by 82 publications

References 36 publications

Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites

Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites

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

The Terpene Mini-Path, a New Promising Alternative for Terpenoids Bio-Production

Contact Info

Product

Resources

About