Construction of a novel Pichia pastoris strain for production of xanthophylls

Araya-Garay, José Miguel; Ageitos, José Manuel; Vallejo, Juán A.; Veiga‐Crespo, Patricia; Sánchez-Pérez, Alicia; Villa, Tomás G.

doi:10.1186/2191-0855-2-24

Cited by 44 publications

(22 citation statements)

References 36 publications

(51 reference statements)

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“…Thus, these engineered hosts are competitive to natural microbial β-carotene producers currently used for production at industrial scale such as the microalga Dunaliella salina which has been reported to synthesize 37.3 mg g −1 DCW per day (García-González et al 2005) or the fungus Blakeslea trispora which could be optimized to produce up to 55 mg g −1 DCW of β-carotene per day (Roukas et al 2015). In contrast, yields of other hosts such as C. glutamicum or P. pastoris are relatively low in the μg g −1 DCW to single-digit mg g −1 DCW range (Araya-Garay et al 2012; Henke et al 2016).…”

Section: Microbial Production Of Different Terpenoid Classesmentioning

confidence: 99%

Identifying and engineering the ideal microbial terpenoid production host

Moser

Pichler

2019

Appl Microbiol Biotechnol

118

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More than 70,000 different terpenoid structures are known so far; many of them offer highly interesting applications as pharmaceuticals, flavors and fragrances, or biofuels. Extraction of these compounds from their natural sources or chemical synthesis is-in many cases-technically challenging with low or moderate yields while wasting valuable resources. Microbial production of terpenoids offers a sustainable and environment-friendly alternative starting from simple carbon sources and, frequently, safeguards high product specificity. Here, we provide an overview on employing recombinant bacteria and yeasts for heterologous de novo production of terpenoids. Currently, Escherichia coli and Saccharomyces cerevisiae are the two best-established production hosts for terpenoids. An increasing number of studies have been successful in engineering alternative microorganisms for terpenoid biosynthesis, which we intend to highlight in this review. Moreover, we discuss the specific engineering challenges as well as recent advances for microbial production of different classes of terpenoids. Rationalizing the current stages of development for different terpenoid production hosts as well as future prospects shall provide a valuable decision basis for the selection and engineering of the cell factory(ies) for industrial production of terpenoid target molecules.

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Section: Microbial Production Of Different Terpenoid Classesmentioning

confidence: 99%

Identifying and engineering the ideal microbial terpenoid production host

Moser

Pichler

2019

Appl Microbiol Biotechnol

118

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“…Araya‐Garay et al. constructed β‐carotene‐producing Pichia strain Pp‐EBIL as described above and used it as a platform strain for further metabolic engineering modifications . Two genes, crtW and crtZ , from Agrobacterium aurantiacum were cloned under the GAP promoter to generate the plasmid pGAPZA‐WZ.…”

Section: Metabolic Engineering Of P Pastorismentioning

confidence: 99%

Heterologous Protein Expression in Pichia pastoris: Latest Research Progress and Applications

Juturu

2017

ChemBioChem

159

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Pichia pastoris is a well‐known platform strain for heterologous protein expression. Over the past five years, different strategies to improve the efficiency of recombinant protein expression by this yeast strain have been developed; these include a patent‐free protein expression kit, construction of the P. pastoris CBS7435Ku70 platform strain with its high efficiency in site‐specific recombination of plasmid DNA into the genomic DNA, the design of synthetic promoters and their variants by combining different core promoters with multiple putative transcription factors, the generation of mutant GAP promoter variants with various promoter strengths, codon optimization, engineering the α‐factor signal sequence by replacing the native glutamic acid at the Kex2 cleavage site with the other 19 natural amino acids and the addition of mammalian signal sequence to the yeast signal sequence, and the co‐expression of single chaperones, multiple chaperones or helper proteins that aid in recombinant protein folding. Publically available high‐quality genome data from multiple strains of P. pastoris GS115, DSMZ 70382, and CBS7435 and the continuous development of yeast expression kits have successfully promoted the metabolic engineering of this strain to produce carotenoids, xanthophylls, nootkatone, ricinoleic acid, dammarenediol‐II, and hyaluronic acid. The cell‐surface display of enzymes has obviously increased enzyme stability, and high‐level intracellular expression of acyl‐CoA and ethanol O‐acyltransferase, lipase and d‐amino acid oxidase has opened up applications in whole‐cell biocatalysis for producing flavor molecules and biodiesel, as well as the deracemization of racemic amino acids. High‐level expression of various food‐grade enzymes, cellulases, and hemicellulases for applications in the food, feed and biorefinery industries is in its infancy and needs strengthening.

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“…The methylotrophic yeast Pichia pastoris (renamed Komagataella phaffii 9 ) is currently used industrially for production of recombinant proteins 10 . Recently, P. pastoris has also been used for production of metabolites including astaxanthin and isobutanol from sugar 11 , 12 . However, relative to model organisms, most native methylotrophs lack the genetic tools and depth of characterisation necessary for the successful metabolic engineering of high-yield and heterologous pathways.…”

Section: Introductionmentioning

confidence: 99%

Adaptive laboratory evolution of native methanol assimilation in Saccharomyces cerevisiae

et al. 2020

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Utilising one-carbon substrates such as carbon dioxide, methane, and methanol is vital to address the current climate crisis. Methylotrophic metabolism enables growth and energy generation from methanol, providing an alternative to sugar fermentation. Saccharomyces cerevisiae is an important industrial microorganism for which growth on one-carbon substrates would be relevant. However, its ability to metabolize methanol has been poorly characterised. Here, using adaptive laboratory evolution and 13C-tracer analysis, we discover that S. cerevisiae has a native capacity for methylotrophy. A systems biology approach reveals that global rearrangements in central carbon metabolism fluxes, gene expression changes, and a truncation of the uncharacterized transcriptional regulator Ygr067cp supports improved methylotrophy in laboratory evolved S. cerevisiae. This research paves the way for further biotechnological development and fundamental understanding of methylotrophy in the preeminent eukaryotic model organism and industrial workhorse, S. cerevisiae.

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Construction of a novel Pichia pastoris strain for production of xanthophylls

Cited by 44 publications

References 36 publications

Identifying and engineering the ideal microbial terpenoid production host

Identifying and engineering the ideal microbial terpenoid production host

Heterologous Protein Expression in Pichia pastoris: Latest Research Progress and Applications

Adaptive laboratory evolution of native methanol assimilation in Saccharomyces cerevisiae

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