Exploiting the Diversity of Saccharomycotina Yeasts To Engineer Biotin-Independent Growth of Saccharomyces cerevisiae

Wronska, Anna K.; Haak, Meinske P; Geraats, Ellen; Slot, Eva Bruins; Broek, Marcel van den; Pronk, Jack T.; Daran, Jean‐Marc

doi:10.1128/aem.00270-20

Cited by 10 publications

(4 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It can be speculated that the amino acid change from serine to proline detected in the other S. cerevisiae strains might generate a non‐functional Cab1p. Intriguingly, the presence of biotin apparently stimulated the growth of the biotin auxotrophic S288C strain, which lacks BIO1 and BIO6 required for biotin biosynthesis (Wronska et al, 2020). Our WG sequencing data revealed that the other three S. cerevisiae strains also lacked BIO1 and BIO6 (Table S7).…”

Section: Resultsmentioning

confidence: 99%

Integrated genomics and phenotype microarray analysis of Saccharomyces cerevisiae industrial strains for rice wine fermentation and recombinant protein production

Son,

Jeon,

Moon

et al. 2023

Microbial Biotechnology

View full text Add to dashboard Cite

The industrial potential of Saccharomyces cerevisiae has extended beyond its traditional use in fermentation to various applications, including recombinant protein production. Herein, comparative genomics was performed with three industrial S. cerevisiae strains and revealed a heterozygous diploid genome for the 98‐5 and KSD‐YC strains (exploited for rice wine fermentation) and a haploid genome for strain Y2805 (used for recombinant protein production). Phylogenomic analysis indicated that Y2805 was closely associated with the reference strain S288C, whereas KSD‐YC and 98‐5 were grouped with Asian and European wine strains, respectively. Particularly, a single nucleotide polymorphism (SNP) in FDC1, involved in the biosynthesis of 4‐vinylguaiacol (4‐VG, a phenolic compound with a clove‐like aroma), was found in KSD‐YC, consistent with its lack of 4‐VG production. Phenotype microarray (PM) analysis showed that KSD‐YC and 98‐5 displayed broader substrate utilization than S288C and Y2805. The SNPs detected by genome comparison were mapped to the genes responsible for the observed phenotypic differences. In addition, detailed information on the structural organization of Y2805 selection markers was validated by Sanger sequencing. Integrated genomics and PM analysis elucidated the evolutionary history and genetic diversity of industrial S. cerevisiae strains, providing a platform to improve fermentation processes and genetic manipulation.

show abstract

Section: Resultsmentioning

confidence: 99%

Integrated genomics and phenotype microarray analysis of Saccharomyces cerevisiae industrial strains for rice wine fermentation and recombinant protein production

Son,

Jeon,

Moon

et al. 2023

Microbial Biotechnology

View full text Add to dashboard Cite

show abstract

“…The GFP dropout plasmid pGGKd017 ( URA3 ) ( Wronska et al, 2020 ) was used as backbone to construct expression cassettes expressing a single 2-oxo acid decarboxylase. As example, the Golden Gate assembly of pGGkp035 ( TDH3pr ), pGGkp182 ( CYC1t ) and pGGKp211 ( Zmpdc1 ) using pGGKd017 as a backbone resulted in the construction of pUDE827 ( URA3, TDH3pr-Zmpdc1-CYC1t ).…”

Section: Methodsmentioning

confidence: 99%

“…The expression cassette bearing YlPDC1 was assembled using Gibson assembly. The gene YlPDC1 (YALI0D10131g, Genome Resources for Yeast Chromosomes database ( https://gryc.inra.fr )) was PCR amplified from genomic DNA of Y. lipolytica W29 ( Magnan et al, 2016 ; Wronska et al, 2020 ) using primers 14187/14188. The TDH3pr and CYC1t were amplified from pGGKp035 and pGGKp182 using primers 14185/14186 and 14189/14190, respectively.…”

Section: Methodsmentioning

confidence: 99%

Elimination of aromatic fusel alcohols as by-products of Saccharomyces cerevisiae strains engineered for phenylpropanoid production by 2-oxo-acid decarboxylase replacement

Hassing

Buijs

Blankerts

et al. 2021

Metabolic Engineering Communications

Self Cite

View full text Add to dashboard Cite

Engineered strains of the yeast Saccharomyces cerevisiae are intensively studied as production platforms for aromatic compounds such as hydroxycinnamic acids, stilbenoids and flavonoids. Heterologous pathways for production of these compounds use l -phenylalanine and/or l -tyrosine, generated by the yeast shikimate pathway, as aromatic precursors. The Ehrlich pathway converts these precursors to aromatic fusel alcohols and acids, which are undesirable by-products of yeast strains engineered for production of high-value aromatic compounds. Activity of the Ehrlich pathway requires any of four S. cerevisiae 2-oxo-acid decarboxylases (2-OADCs): Aro10 or the pyruvate-decarboxylase isoenzymes Pdc1, Pdc5, and Pdc6. Elimination of pyruvate-decarboxylase activity from S. cerevisiae is not straightforward as it plays a key role in cytosolic acetyl-CoA biosynthesis during growth on glucose. In a search for pyruvate decarboxylases that do not decarboxylate aromatic 2-oxo acids, eleven yeast and bacterial 2-OADC-encoding genes were investigated. Homologs from Kluyveromyces lactis ( KlPDC1 ), Kluyveromyces marxianus ( KmPDC1 ), Yarrowia lipolytica ( YlPDC1 ), Zymomonas mobilis ( Zmpdc1 ) and Gluconacetobacter diazotrophicus ( Gdpdc1.2 and Gdpdc1.3 ) complemented a Pdc − strain of S. cerevisiae for growth on glucose. Enzyme-activity assays in cell extracts showed that these genes encoded active pyruvate decarboxylases with different substrate specificities. In these in vitro assays, Zm Pdc1, Gd Pdc1.2 or Gd Pdc1.3 had no substrate specificity towards phenylpyruvate. Replacing Aro10 and Pdc1,5,6 by these bacterial decarboxylases completely eliminated aromatic fusel-alcohol production in glucose-grown batch cultures of an engineered coumaric acid-producing S. cerevisiae strain. These results outline a strategy to prevent formation of an important class of by-products in ‘chassis’ yeast strains for production of non-native aromatic compounds.

show abstract

“…Recent research indicates that also the first, unresolved step of biotin biosynthesis in yeasts, catalysed by Bio1 orthologs, is oxygen dependent (Wronska et al . 2020 ). Although heme biosynthesis is oxygen dependent, heme-containing proteins in S. cerevisiae are strongly associated with aerobic metabolism.…”

Section: Introductionmentioning

confidence: 99%

Critical parameters and procedures for anaerobic cultivation of yeasts in bioreactors and anaerobic chambers

Mooiman

Bouwknegt

Dekker

et al. 2021

FEMS Yeast Research

View full text Add to dashboard Cite

All known facultatively fermentative yeasts require molecular oxygen for growth. Only in a small number of yeast species, these requirements can be circumvented by supplementation of known anaerobic growth factors such as nicotinate, sterols and unsaturated fatty acids. Biosynthetic oxygen requirements of yeasts are typically small and, unless extensive precautions are taken to minimize inadvertent entry of trace amounts of oxygen, easily go unnoticed in small-scale laboratory cultivation systems. This paper discusses critical points in the design of anaerobic yeast cultivation experiments in anaerobic chambers and laboratory bioreactors. Serial transfer or continuous cultivation to dilute growth factors present in anaerobically pre-grown inocula, systematic inclusion of control strains and minimizing the impact of oxygen diffusion through tubing are identified as key elements in experimental design. Basic protocols are presented for anaerobic-chamber and bioreactor experiments.

show abstract

Exploiting the Diversity of Saccharomycotina Yeasts To Engineer Biotin-Independent Growth of Saccharomyces cerevisiae

Cited by 10 publications

References 69 publications

Integrated genomics and phenotype microarray analysis of Saccharomyces cerevisiae industrial strains for rice wine fermentation and recombinant protein production

Integrated genomics and phenotype microarray analysis of Saccharomyces cerevisiae industrial strains for rice wine fermentation and recombinant protein production

Elimination of aromatic fusel alcohols as by-products of Saccharomyces cerevisiae strains engineered for phenylpropanoid production by 2-oxo-acid decarboxylase replacement

Critical parameters and procedures for anaerobic cultivation of yeasts in bioreactors and anaerobic chambers

Contact Info

Product

Resources

About