A Copper-Regulated Yarrowia lipolytica Vector for the Expression of Heterologous Proteins

Domínguez, Ángel; Prado, Marciano; García, Sara Mallén

doi:10.1007/978-3-642-55758-3_59

Cited by 2 publications

(4 citation statements)

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“…Cu 2+ -inducible promoter P CUP1 from gene encoding metallothionein, which has low molecular weight, cysteine-rich protein being capable of binding heavy metals such as copper, zinc, selenium, cadmium, mercury, and silver, has been identified in yeast S. cerevisiae . Similarly, it indicated that two promoters in Y. lipolytica E150 could be used to express genes under induction of copper . However, there were neither sequences provided, nor activities reported.…”

Section: Resultsmentioning

confidence: 99%

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Expanding Toolbox for Genes Expression of Yarrowia lipolytica to Include Novel Inducible, Repressible, and Hybrid Promoters

Xiong

Chen

2020

ACS Synth. Biol.

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Promoters are critical tools to precisely control gene expression for both synthetic biology and metabolic engineering.Although Yarrowia lipolytica has demonstrated many industrially relevant advantages, promoter discovery efforts on this nonconventional yeast are limited due to the challenge in finding suitable inducible and repressible promoters. Six copper-inducible promoters and five repressible promoters were isolated in this work. Especially, Cu 2+ -repressible promoters showed relatively high activity under non-repressing conditions compared with a constitutive promoter, but the strength could be almost fully repressed by a supplement of a low content of Cu 2+ . The six Cu 2+ -inducible promoters were engineered to improve their dynamic regulation range with a tandem upstream activation sequence. An engineered promoter was successfully used to construct a more productive pathway for production of a novel bioproduct, wax ester, than that used for both Cu 2+ -inducible promoter and constitutive promoter. This study provides effective tools applicable to fine-tune the gene expression in this microbial host.

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Section: Resultsmentioning

confidence: 99%

“…21 Similarly, it indicated that two promoters in Y. lipolytica E150 could be used to express genes under induction of copper. 22 However, there were neither sequences provided, nor activities reported. In this study, six genes namely MT-1 to MT-6 encoding metallothionein were retrieved from Y. lipolytica genome (Figure 1A).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Expanding Toolbox for Genes Expression of Yarrowia lipolytica to Include Novel Inducible, Repressible, and Hybrid Promoters

Xiong

Chen

2020

ACS Synth. Biol.

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“…The autosomal vector pMM4 was used to express the Alt a 1 allergen. The YlMETII promoter was obtained from plasmid pSG70 (García, ; Domínguez et al ., ) and cloned between the EcoRI‐BamHI restriction sites of the pBluescript‐SK. The Alt a 1‐coding gene sequence (Asturias et al ., ) was cloned after this promoter into the BamHI site, resulting in the pMMR2 vector.…”

Section: Methodsmentioning

confidence: 99%

“…Several Y. lipolytica promoters have been used for recombinant protein expression (Domínguez et al, 1998;Muller et al, 1998;Wang et al, 1999;Pignède et al, 2000). The copper-inducible bi-directional promoter of YlMTPI and YlMTPII genes has been characterized previously (García, 1993;Domínguez et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Expression of Alt a 1 allergen from Alternaria alternata in the yeast Yarrowia lipolytica

Morín

Asturias²,

Domínguez

2012

FEMS Microbiol Lett

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Allergies affect almost 25% of the population in industrialized countries. Alternaria alternata is known to be a significant source of aeroallergens and sensitization to this mold is a risk factor for the development of wheezing, asthma, and atopic dermatitis. Diagnosis and treatment of allergies requires the production of large amounts of pure and well defined protein. Yarrowia lipolytica, a non-pathogenic ascomycete able to secrete high levels of enzymes that can grow in inexpensive substrates, has been considered a useful host for heterologous gene expression. In the present work, we have developed two vectors for expressing Alt a 1, the most relevant A. alternata allergen, in Y. lipolytica. One vector is autosomal and one is integrative. With both systems, rAlt a 1 was secreted into the culture medium. The immunological characteristics of the purified recombinant allergen were determined by IgE-blot using sera from 42 A. alternata-allergic patients. We have carried out ELISA-inhibition experiments using sera from four patients to compare the IgE-binding capacity of natural and recombinant allergens. Our results show that Y. lipolytica is able to produce a recombinant Alt a 1 which is immunochemically equivalent to the natural counterpart and could be used for immunotherapy and diagnostics.

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A Copper-Regulated Yarrowia lipolytica Vector for the Expression of Heterologous Proteins

Cited by 2 publications

References 5 publications

Expanding Toolbox for Genes Expression of Yarrowia lipolytica to Include Novel Inducible, Repressible, and Hybrid Promoters

Expanding Toolbox for Genes Expression of Yarrowia lipolytica to Include Novel Inducible, Repressible, and Hybrid Promoters

Expression of Alt a 1 allergen from Alternaria alternata in the yeast Yarrowia lipolytica

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