Faculty Opinions recommendation of Origin of Lactose Fermentation in Kluyveromyces lactis by Interspecies Transfer of a Neo-functionalized Gene Cluster during Domestication.

2019 ◽  
Author(s):  
Aaron Neiman
Keyword(s):  
Gene Cluster ◽  
Kluyveromyces Lactis ◽  
Lactose Fermentation

scholarly journals Origin of Lactose Fermentation in Kluyveromyces lactis by Interspecies Transfer of a Neo-functionalized Gene Cluster during Domestication

2019 ◽  
Vol 29 (24) ◽  
pp. 4284-4290.e2 ◽  
Author(s):  
Javier A. Varela ◽  
Martina Puricelli ◽  
Raúl A. Ortiz-Merino ◽  
Romina Giacomobono ◽  
Stephanie Braun-Galleani ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Kluyveromyces Lactis ◽  
Lactose Fermentation

scholarly journals Origin of Lactose Fermentation in Kluyveromyces lactis by Interspecies Transfer of a Neofunctionalized Gene Cluster During Domestication

2019 ◽  
Author(s):  
Javier A. Varela ◽  
Martina Puricelli ◽  
Raúl A. Ortiz-Merino ◽  
Romina Giacomobono ◽  
Stephanie Braun-Galleani ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Kluyveromyces Lactis ◽  
Lactose Fermentation

scholarly journals Adaptive Evolution of a Lactose-Consuming Saccharomyces cerevisiae Recombinant

2008 ◽  
Vol 74 (6) ◽  
pp. 1748-1756 ◽  
Author(s):  
Pedro M. R. Guimarães ◽  
Jean François ◽  
Jean Luc Parrou ◽  
José A. Teixeira ◽  
Lucília Domingues
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Copy Number ◽  
Cheese Whey ◽  
Kluyveromyces Lactis ◽  
Strain Improvement ◽  
Attractive Alternative ◽  
Evolutionary Engineering ◽  
Serial Transfer ◽  
Lactose Fermentation ◽  
Transcriptional Induction

ABSTRACT The construction of Saccharomyces cerevisiae strains that ferment lactose has biotechnological interest, particularly for cheese whey fermentation. A flocculent lactose-consuming S. cerevisiae recombinant expressing the LAC12 (lactose permease) and LAC4 (β-galactosidase) genes of Kluyveromyces lactis was constructed previously but showed poor efficiency in lactose fermentation. This strain was therefore subjected to an evolutionary engineering process (serial transfer and dilution in lactose medium), which yielded an evolved recombinant strain that consumed lactose twofold faster, producing 30% more ethanol than the original recombinant. We identified two molecular events that targeted the LAC construct in the evolved strain: a 1,593-bp deletion in the intergenic region (promoter) between LAC4 and LAC12 and a decrease of the plasmid copy number by about 10-fold compared to that in the original recombinant. The results suggest that the intact promoter was unable to mediate the induction of the transcription of LAC4 and LAC12 by lactose in the original recombinant and that the deletion established the transcriptional induction of both genes in the evolved strain. We propose that the tuning of the expression of the heterologous LAC genes in the evolved recombinant was accomplished by the interplay between the decreased copy number of both genes and the different levels of transcriptional induction for LAC4 and LAC12 resulting from the changed promoter structure. Nevertheless, our results do not exclude other possible mutations that may have contributed to the improved lactose fermentation phenotype. This study illustrates the usefulness of simple evolutionary engineering approaches in strain improvement. The evolved strain efficiently fermented threefold-concentrated cheese whey, providing an attractive alternative for the fermentation of lactose-based media.

Inter-Strain Hybridization of Kluyveromyces lactis for creating efficient lactose-fermenting Yeast

2021 ◽  
Vol 37 (4) ◽  
pp. 43-50
Author(s):  
E.S. Naumova
Keyword(s):  
Kluyveromyces Lactis ◽  
Molecular Genetic ◽  
Genetic Research ◽  
Amino Acid Sequences ◽  
Research Center ◽  
Lactose Permease ◽  
Kurchatov Institute ◽  
Molecular Genetic Study ◽  
Lactose Fermentation ◽  
Physiological Tests

A molecular genetic study of Kluyveromyces lactis yeasts isolated from various dairy products in the countries of the former Soviet Union and other regions of the world has been carried out. Based on physiological tests, four strains were selected that carry different LAC loci and are characterized by good fermentation intensity: VKM Y-1339 (LAC3), VKM Y-1333 (LAC3), NRRL Y-1118 (LAC1), and NRRL Y-1140 (LAC2). Eleven hybrids of the selected strains with different rates of lactose fermentation were obtained. No correlation was found between the intensity of lactose fermentation and the amino acid sequences of the LAC12 lactose permease gene of the LAC1, LAC2, and LAC3 loci. Apparently, a specific combination of genotypes of crossed strains has a more significant effect on the fermentation activity. The results obtained showed that inter-strain hybridization of K. lactis dairy yeast is an effective method for creating new strains with high fermentation capacity. Hybrids H2-3 (NRRL Y-1118 × VKM Y-1333) and H3-3 (NRRL Y-1140 × VKM Y-1333) with the highest ability to ferment lactose are of interest for further molecular genetic research and breeding programs. Key words: Kluyveromyces lactis, β-galactosidase, lactose permease, LAC4, LAC12, LAC1 locus, LAC2 locus, LAC3 locus, inter-strain hybridization, lactose fermentation, heterosis Acknowledgment - The authors are grateful to the Genomic Center of the Kurchatov Institute SRC---GosNIIgentika for sequencing the nucleotide sequences of the LAC12 genes for lactose permease on the Applied Biosystems 3730 automated analyzer. Funding - This work was supported by an internal grant from the National Research Center Kurchatov Institute (order of the National Research Center Kurchatov Institute No. 1779).

scholarly journals Functional and evolutionary characterization of a secondary metabolite gene cluster in budding yeasts

2018 ◽  
Vol 115 (43) ◽  
pp. 11030-11035 ◽  
Author(s):  
David J. Krause ◽  
Jacek Kominek ◽  
Dana A. Opulente ◽  
Xing-Xing Shen ◽  
Xiaofan Zhou ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Gene Cluster ◽  
Kluyveromyces Lactis ◽  
Phylogenetic Analyses ◽  
Iron Binding ◽  
Acid Biosynthesis ◽  
Transcription Factor Genes ◽  
Domains Of Life ◽  
Secondary Metabolite Gene Cluster

Secondary metabolites are key in how organisms from all domains of life interact with their environment and each other. The iron-binding molecule pulcherrimin was described a century ago, but the genes responsible for its production in budding yeasts have remained uncharacterized. Here, we used phylogenomic footprinting on 90 genomes across the budding yeast subphylum Saccharomycotina to identify the gene cluster associated with pulcherrimin production. Using targeted gene replacements in Kluyveromyces lactis, we characterized the four genes that make up the cluster, which likely encode two pulcherriminic acid biosynthesis enzymes, a pulcherrimin transporter, and a transcription factor involved in both biosynthesis and transport. The requirement of a functional putative transporter to utilize extracellular pulcherrimin-complexed iron demonstrates that pulcherriminic acid is a siderophore, a chelator that binds iron outside the cell for subsequent uptake. Surprisingly, we identified homologs of the putative transporter and transcription factor genes in multiple yeast genera that lacked the biosynthesis genes and could not make pulcherrimin, including the model yeast Saccharomyces cerevisiae. We deleted these previously uncharacterized genes and showed they are also required for pulcherrimin utilization in S. cerevisiae, raising the possibility that other genes of unknown function are linked to secondary metabolism. Phylogenetic analyses of this gene cluster suggest that pulcherrimin biosynthesis and utilization were ancestral to budding yeasts, but the biosynthesis genes and, subsequently, the utilization genes, were lost in many lineages, mirroring other microbial public goods systems that lead to the rise of cheater organisms.

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