Genetic engineering of nonconventional yeasts for the production of valuable compounds

Sibirny, Andriy А.; Madzak, Catherine; Fickers, Patrick

doi:10.1201/b17587-5

Cited by 8 publications

(18 citation statements)

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“…Non-conventional model yeast species such as Pichia pastoris , Hansuluna polymorpha , and Yarrowia lipolytica have received much attention in recent years because they can serve as cell factories for producing heterologous proteins at lab or industrial scales [ 1 , 2 ]. High yields of more than one hundred heterologous proteins have already been successfully obtained using Y. lipolytica, underscoring this yeast’s production potential [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

Deciphering how LIP2 and POX2 promoters can optimally regulate recombinant protein production in the yeast Yarrowia lipolytica

et al. 2016

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BackgroundIn recent years, the non-conventional model yeast species Yarrowia lipolytica has received much attention because it is a useful cell factory for producing recombinant proteins. In this species, expression vectors involving LIP2 and POX2 promoters have been developed and used successfully for protein production at yields similar to or even higher than those of other cell factories, such as Pichia pastoris. However, production processes involving these promoters can be difficult to manage, especially if carried out at large scales in fed-batch bioreactors, because they require hydrophobic inducers, such as oleic acid or methyl oleate. Thus, the challenge has become to reduce loads of hydrophobic substrates while simultaneously promoting recombinant protein production. One possible solution is to replace a portion of the inducer with a co-substrate that can serve as an alternative energy source. However, implementing such an approach would require detailed knowledge of how carbon sources impact promoter regulation, which is surprisingly still lacking for the LIP2 and POX2 promoters. This study’s aim was thus to better characterize promoter regulation and cell metabolism in Y. lipolytica cultures grown in media supplemented with different carbon sources.ResultspPOX2 induction could be detected when glucose or glycerol was used as sole carbon source, which meant these carbon source could not prevent promoter induction. In addition, when a mixture of glucose and oleic acid was used in complex medium, pPOX2 induction level was lower that that of pLIP2. In contrast, pLIP2 induction was absent when glucose was present in the culture medium, which meant that cell growth could occur without any recombinant gene expression. When a 40/60 mixture of glucose and oleic acid (w/w) was used, a tenfold increase in promoter induction, as compared to when an oleic-acid-only medium was observed. It was also clear that individual cells were adapting metabolically to use both glucose and oleic acid. Indeed, no distinct subpopulations that specialized on glucose versus oleic acid were observed; such an outcome would have led to producer and non-producer phenotypes. In medium containing both glucose and oleic acid, cells tended to directly metabolize oleic acid instead of storing it in lipid bodies.ConclusionsThis study found that pLIP2 is a promoter of choice as compared to pPOX2 to drive gene expression for recombinant protein production by Y. lipolytica used as cell factory.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0558-8) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Deciphering how LIP2 and POX2 promoters can optimally regulate recombinant protein production in the yeast Yarrowia lipolytica

et al. 2016

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“…This matter of fact could be attributed at least in part to social acceptance issues concerning GM microorganisms, especially in the domain of food applications. Until now, only a few commercial or industrial applications of GM Y. lipolytica strains can be reported [1,37]. GM Y. lipolytica cell factories are presently used for industrial production of two kinds of food/feed additives: carotenoids and polyunsaturated fatty acids (PUFAs)-rich SCOs.…”

Section: Commercial Applications Of Genetically Modified Strainsmentioning

confidence: 99%

“…GM Y. lipolytica cell factories are presently used for industrial production of two kinds of food/feed additives: carotenoids and polyunsaturated fatty acids (PUFAs)-rich SCOs. The technology of carotenoids production by Y. lipolytica was developed by Microbia (USA) and then acquired by DSM (Heerlen, Netherlands) [37]. The technology of PUFAsrich SCOs production by a heavily engineered Y. lipolytica strain derived from the ATCC 20362 wild-type isolate (cf.…”

Section: Commercial Applications Of Genetically Modified Strainsmentioning

confidence: 99%

“…The high potential of Y. lipolytica for industrial applications has been exploited since more than seventies years, at first in the fields of biomass and valuable metabolites production, using proprietary wild-type isolates or traditionally improved strains (mutants, strains issued from hybridizations and crossings) [1,36,37]. In the 1950s, and until the oil crisis of the 1970s, the British Petroleum Company (BP, London, UK) applied the alkanedegrading capacities of this yeast to the production of single-cell protein (SCP) from crude oil.…”

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confidence: 99%

“…In the 1970s, Pfizer Inc. (New York City, USA) applied Y. lipolytica to industrial citric acid production. This technology was repurchased in the 1990s by Archer Daniels Midland Company (ADM, Chicago, IL, USA), who is still (to the best of our knowledge) producing some citric acid from Y. lipolytica, using mostly corn or rape seed oil as substrates [1,36,37]. Other nowadays applications of proprietary wild-type or traditionally improved Y. lipolytica strains include, in the field of food industry, erythritol production by Baolingbao Biology Co. (Yucheng, Shandong, China) and use of Y. lipolytica biomass as fodder yeast for farm and pet animals by Skotan SA (Chorzów, Poland), who also develop prebiotic/probiotic applications [1,38].…”

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Yarrowia lipolytica strains and their biotechnological applications: how natural biodiversity and metabolic engineering could contribute to cell factories improvement

Madzak

2021

Preprint

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Among non-conventional yeasts of industrial interest, the dimorphic oleaginous yeast Yarrowia lipolytica appears as one of the most attractive for a large range of white biotechnology applications, from heterologous proteins secretion to cell factories process development. The past, present and potential applications of wild type, traditionally improved or genetically modified Yarrowia lipolytica strains will be resumed, together with the wide array of molecular tools now available to genetically engineer and metabolically remodel this yeast. The present review will also provide a detailed description of Yarrowia lipolytica strains and highlight the natural biodiversity of this yeast, a subject little touched upon in most previous reviews. This work intends to fill this gap by retracing the genealogy of the main Yarrowia lipolytica strains of industrial interest, by illustrating the search for new genetic backgrounds and by providing data about the main publicly available strains in yeast collections worldwide. At last, it will focus on exemplifying how advances in engineering tools can leverage a better biotechnological exploitation of the natural biodiversity of Yarrowia lipolytica and of other yeasts from the Yarrowia clade.

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Yarrowia lipolytica: recent achievements in heterologous protein expression and pathway engineering

Madzak

2015

Appl Microbiol Biotechnol

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The oleaginous yeast Yarrowia lipolytica has become a recognized system for expression/secretion of heterologous proteins. This non-conventional yeast is currently being developed as a workhorse for biotechnology by several research groups throughout the world, especially for single-cell oil production, whole cell bioconversion and upgrading of industrial wastes. This mini-review presents established tools for protein expression in Y. lipolytica and highlights novel developments in the areas of promoter design, surface display, and host strain or metabolic pathway engineering. An overview of the industrial and commercial biotechnological applications of Y. lipolytica is also presented.

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Genetic engineering of nonconventional yeasts for the production of valuable compounds

Cited by 8 publications

References 132 publications

Deciphering how LIP2 and POX2 promoters can optimally regulate recombinant protein production in the yeast Yarrowia lipolytica

Deciphering how LIP2 and POX2 promoters can optimally regulate recombinant protein production in the yeast Yarrowia lipolytica

Yarrowia lipolytica strains and their biotechnological applications: how natural biodiversity and metabolic engineering could contribute to cell factories improvement

Yarrowia lipolytica: recent achievements in heterologous protein expression and pathway engineering

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