Engineering Saccharomyces cerevisiae for efficient production of recombinant proteins

Yang, Shuo; Song, Liyun; Wang, Jing; Zhao, Jianzhi; Tang, Hongting; Bao, Xiaoming

doi:10.1016/j.engmic.2023.100122

Cited by 7 publications

(3 citation statements)

References 138 publications

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“…Different strategies have been used to improve recombinant protein production in S. cerevisiae ; among them are the engineering of galactose inducible promoters (P GAL1 , P GAL2 , P GAL7 and P GAL 10 ), codon optimization, and enhancement of the protein secretion system. 42 In this work, it was shown that peptide signals found in LIP3 and YJR107W genes from K. marxianus are recognized by S. cerevisiae since the secretion of the potential encoded recombinant proteins, and their engineering could enhance the extracellular enzyme levels induced by some efficient promoter. In a study, several combinations of mutations in the GAL10 promoter of S. cerevisiae were used to test their efficiency in regulating the Candida antarctiva CalB gene, which encodes for lipase B.…”

Section: Resultsmentioning

confidence: 97%

Lipase activity of recombinant KmYJR107Wp and KmLIP3p enzymes expressed in Saccharomyces cerevisiae BY4742 from Kluyveromyces marxianus L2029

Martínez-Corona,

Canizal-García,

Madrigal-Perez

et al. 2024

Journal of Genetic Engineering and Biotechnology

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Section: Resultsmentioning

confidence: 97%

Lipase activity of recombinant KmYJR107Wp and KmLIP3p enzymes expressed in Saccharomyces cerevisiae BY4742 from Kluyveromyces marxianus L2029

Martínez-Corona,

Canizal-García,

Madrigal-Perez

et al. 2024

Journal of Genetic Engineering and Biotechnology

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“…Genotypic and phenotypic characteristics of Saccharomyces must be considered carefully to develop live vectors using the yeasts for the delivery of functional therapeutic proteins. For instance, codon optimization considering the codon bias of Saccharomyces is a critical prerequisite to maximize the efficiency of the production and secretion of functional proteins by Saccharomyces [66,75]. The secretion signal is another major factor affecting protein delivery efficiency.…”

Section: In Situ Delivery Of Therapeutic Proteinsmentioning

confidence: 99%

Therapeutic Applications of Native and Engineered Saccharomyces Yeasts

Kwak

2024

Fermentation

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Saccharomyces cerevisiae var. boulardii (Sb) is currently receiving significant attention as a synthetic probiotic platform due to its ease of manipulation and inherent effectiveness in promoting digestive health. A comprehensive exploration of Sb and other S. cerevisiae strains (Sc) would shed light on the refinement and expansion of their therapeutic applications. This review aims to provide a thorough overview of Saccharomyces yeasts from their native health benefits to recent breakthroughs in the engineering of Saccharomyces yeasts as synthetic therapeutic platforms. Molecular typing and phenotypic assessments have uncovered notable distinctions, including the superior thermotolerance and acid tolerance exhibited by Sb, which are crucial attributes for probiotic functions. Moreover, parabiotic and prebiotic functionalities originating from yeast cell wall oligosaccharides have emerged as pivotal factors influencing the health benefits associated with Sb and Sc. Consequently, it has become imperative to select an appropriate yeast strain based on a comprehensive understanding of its actual action in the gastrointestinal tract and the origins of the targeted advantages. Overall, this review underscores the significance of unbiased and detailed comparative studies for the judicious selection of strains.

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“…Some natural product biosynthesis enzymes (such as polyketide synthases and cytochrome P450s) often suffer from low catalytic activities, which should be expressed at high levels to maximize metabolic flux towards the desired products. Traditionally, gene dosage amplification is a commonly employed strategy to boost foreign gene expression levels. − Considering the instability of episomal plasmids in P. pastoris , multicopy genome integration should be performed for gene dosage amplification. As the traditional stepwise integration strategy is time-consuming and labor-intensive, there is an urgent need to implement multicopy integration technology to construct stable and high-expression strains efficiently.…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Mediated rDNA Integration and Fluorescence Screening for Pathway Optimization in Pichia pastoris

Jiang,

Li,

Wang

et al. 2024

Chem Bio Eng.

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Gene dosage amplification is an effective strategy to improve the performance of heterologous genes and pathways. Pichia pastoris is an excellent recombinant protein expression host with high efficiency in protein folding and glycosylation. However, the traditional iterative multicopy integration method typically faces challenges such as being time-consuming and having high cost and potential gene mutations. Accordingly, we established CRISPR-mediated rDNA integration and fluorescence screening for pathway optimization (CRISPO) for multicopy pathway integration in a single-step and antibiotic-free manner. With geraniol biosynthesis as a case study, we designed CRISPO based on the use of glycerol-induced and glucose-repressed promoters (CRISPOi) or strong constitutive promoters (CRISPOc) to drive the expression of the red fluorescent protein mCherry as the screening marker. We employed CRISPOi for stable strain construction by multicopy integration of the geraniol synthase encoding gene, achieving a 19.5-fold increase in geraniol production. We demonstrated CRISPOc for visualizing and determining the ratelimiting steps of the mevalonate pathway, with HMG1 and ERG12 identified as the major rate-limiting enzymes through two rounds of exploration. Ultimately, CRISPO enabled us to construct an engineered P. pastoris strain producing 1.66 g/L geraniol (with a total of 2.12 g/L monoterpenoids) and 6.27 g/L geraniol (with a total of 6.48 g/L monoterpenoids) in 24-well plates and 5 L fermenters, respectively, representing the highest titer and productivity of geraniol ever reported. CRISPO is an important addition to the synthetic biology toolbox for the construction and optimization of P. pastoris cell factories.

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Engineering Saccharomyces cerevisiae for efficient production of recombinant proteins

Cited by 7 publications

References 138 publications

Lipase activity of recombinant KmYJR107Wp and KmLIP3p enzymes expressed in Saccharomyces cerevisiae BY4742 from Kluyveromyces marxianus L2029

Lipase activity of recombinant KmYJR107Wp and KmLIP3p enzymes expressed in Saccharomyces cerevisiae BY4742 from Kluyveromyces marxianus L2029

Therapeutic Applications of Native and Engineered Saccharomyces Yeasts

CRISPR-Mediated rDNA Integration and Fluorescence Screening for Pathway Optimization in Pichia pastoris

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