scholarly journals Functional genomics of lipid metabolism in the oleaginous yeast Rhodosporidium toruloides

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Samuel T Coradetti ◽  
Dominic Pinel ◽  
Gina M Geiselman ◽  
Masakazu Ito ◽  
Stephen J Mondo ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Functional Genomics ◽  
Lipid Accumulation ◽  
Protein Modification ◽  
Carbon Sources ◽  
Acid Synthesis ◽  
Rhodosporidium Toruloides ◽  
Chemical Production ◽  
Amino Acid Synthesis ◽  
High Concentrations

The basidiomycete yeast Rhodosporidium toruloides (also known as Rhodotorula toruloides) accumulates high concentrations of lipids and carotenoids from diverse carbon sources. It has great potential as a model for the cellular biology of lipid droplets and for sustainable chemical production. We developed a method for high-throughput genetics (RB-TDNAseq), using sequence-barcoded Agrobacterium tumefaciens T-DNA insertions. We identified 1,337 putative essential genes with low T-DNA insertion rates. We functionally profiled genes required for fatty acid catabolism and lipid accumulation, validating results with 35 targeted deletion strains. We identified a high-confidence set of 150 genes affecting lipid accumulation, including genes with predicted function in signaling cascades, gene expression, protein modification and vesicular trafficking, autophagy, amino acid synthesis and tRNA modification, and genes of unknown function. These results greatly advance our understanding of lipid metabolism in this oleaginous species and demonstrate a general approach for barcoded mutagenesis that should enable functional genomics in diverse fungi.

scholarly journals Functional genomics of lipid metabolism in the oleaginous yeast Rhodosporidium toruloides

2017 ◽  
Author(s):  
Samuel T Coradetti ◽  
Dominic Pinel ◽  
Gina Geiselman ◽  
Masakazu Ito ◽  
Stephen Mondo ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Lipid Metabolism ◽  
Functional Genomics ◽  
Lipid Accumulation ◽  
Protein Modification ◽  
Carbon Sources ◽  
Acid Synthesis ◽  
Rhodosporidium Toruloides ◽  
Chemical Production ◽  
Amino Acid Synthesis

AbstractThe basidomycete yeast Rhodosporidium toruloides (a.k.a. Rhodotorula toruloides) accumulates high concentrations of lipids and carotenoids from diverse carbon sources. It has great potential as a model for the cellular biology of lipid droplets and for sustainable chemical production. We developed a method for high-throughput genetics (RB-TDNAseq), using sequence-barcoded Agrobacterium tumefaciens T-DNA insertions into the R. toruloides genome. We identified 1337 putative essential genes with low T-DNA insertion rates. We functionally profiled genes required for fatty acid catabolism and lipid accumulation, validating results with 35 targeted deletion strains. We found that both mitochondrial and peroxisomal enzymes were required for growth on fatty acids, with different peroxisomal enzymes required on different fatty acids. We identified a high-confidence set of 150 genes affecting lipid accumulation, including genes with predicted function in signaling cascades, gene expression, protein modification and vesicular trafficking, autophagy, amino acid synthesis and tRNA modification, as well as genes of unknown function. These results greatly advance our understanding of lipid metabolism in this oleaginous species, identify key biological processes to be further explored and optimized for production of lipid-based bioproducts, and demonstrate a general approach for barcoded mutagenesis that should enable functional genomics in diverse fungi.

scholarly journals Effect of Pyruvate Carboxylase Overexpression on the Physiology of Corynebacterium glutamicum

2002 ◽  
Vol 68 (11) ◽  
pp. 5422-5428 ◽  
Author(s):  
Mattheos A. G. Koffas ◽  
Gyoo Yeol Jung ◽  
Juan C. Aon ◽  
Gregory Stephanopoulos
Keyword(s):  
Corynebacterium Glutamicum ◽  
Pyruvate Carboxylase ◽  
Carbon Sources ◽  
Acid Synthesis ◽  
Aspartate Kinase ◽  
Growth Enhancement ◽  
Lysine Production ◽  
Carboxylase Activity ◽  
Amino Acid Synthesis ◽  
Permeabilized Cells

ABSTRACT Pyruvate carboxylase was recently sequenced in Corynebacterium glutamicum and shown to play an important role of anaplerosis in the central carbon metabolism and amino acid synthesis of these bacteria. In this study we investigate the effect of the overexpression of the gene for pyruvate carboxylase (pyc) on the physiology of C. glutamicum ATCC 21253 and ATCC 21799 grown on defined media with two different carbon sources, glucose and lactate. In general, the physiological effects of pyc overexpression in Corynebacteria depend on the genetic background of the particular strain studied and are determined to a large extent by the interplay between pyruvate carboxylase and aspartate kinase activities. If the pyruvate carboxylase activity is not properly matched by the aspartate kinase activity, pyc overexpression results in growth enhancement instead of greater lysine production, despite its central role in anaplerosis and aspartic acid biosynthesis. Aspartate kinase regulation by lysine and threonine, pyruvate carboxylase inhibition by aspartate (shown in this study using permeabilized cells), as well as well-established activation of pyruvate carboxylase by lactate and acetyl coenzyme A are the key factors in determining the effect of pyc overexpression on Corynebacteria physiology.

scholarly journals Heterologous Expression of Two Malate Transporters From an Oleaginous Fungus Mucor circinelloides Improved the Lipid Accumulation in Mucor lusitanicus

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiuwen Wang ◽  
Hassan Mohamed ◽  
Yonghong Bao ◽  
Chen Wu ◽  
Wenyue Shi ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Lipid Accumulation ◽  
Carbon Sources ◽  
Reducing Power ◽  
Acid Synthesis ◽  
Mucor Circinelloides ◽  
Microbial Lipids ◽  
Intracellular Lipid ◽  
Engineering Technology ◽  
Oleaginous Fungus

The fungus, Mucor lusitanicus, is of great interest for microbial lipids, because of its ability to accumulate intracellular lipid using various carbon sources. The biosynthesis of fatty acid requires the reducing power NADPH, and acetyl-CoA, which is produced by the cleavage of citrate in cytosol. In this study, we employed different strategies to increase lipid accumulation in the low lipid-producing fungi via metabolic engineering technology. Hence, we constructed the engineered strain of M. lusitanicus CBS 277.49 by using malate transporter (mt) and 2-oxoglutarate: malate antiporter (sodit) from M. circinelloides WJ11. In comparison with the control strain, the lipid content of the overexpressed strains of mt and sodit genes were increased by 24.6 and 33.8%, respectively. These results showed that mt and sodit can affect the distribution of malate in mitochondria and cytosol, provide the substrates for the synthesis of citrate in the mitochondria, and accelerate the transfer of citrate from mitochondria to cytosol, which could play a significant regulatory role in fatty acid synthesis leading to lipids over accumulation.

scholarly journals Author response: Functional genomics of lipid metabolism in the oleaginous yeast Rhodosporidium toruloides

2018 ◽  
Author(s):  
Samuel T Coradetti ◽  
Dominic Pinel ◽  
Gina M Geiselman ◽  
Masakazu Ito ◽  
Stephen J Mondo ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Functional Genomics ◽  
Oleaginous Yeast ◽  
Rhodosporidium Toruloides ◽  
Author Response

Influence of increased nutrient supply on Microcystis aeruginosa at cellular and proteomic levels

2020 ◽  
Vol 85 ◽  
pp. 47-58
Author(s):  
Y Jiang ◽  
Y Liu
Keyword(s):  
Microcystis Aeruginosa ◽  
Regulatory Protein ◽  
Acid Synthesis ◽  
Nutrient Supply ◽  
Nitrogen Supply ◽  
Nitrogen And Phosphorus ◽  
Comprehensive Description ◽  
Amino Acid Synthesis ◽  
High Nutrient ◽  
Proteomic Responses

Various studies have observed that increased nutrient supply promotes the growth of bloom-forming cyanobacteria, but only a limited number of studies have investigated the influence of increased nutrient supply on bloom-forming cyanobacteria at the proteomic level. We investigated the cellular and proteomic responses of Microcystis aeruginosa to elevated nitrogen and phosphorus supply. Increased supply of both nutrients significantly promoted the growth of M. aeruginosa and the synthesis of chlorophyll a, protein, and microcystins. The release of microcystins and the synthesis of polysaccharides negatively correlated with the growth of M. aeruginosa under high nutrient levels. Overexpressed proteins related to photosynthesis, and amino acid synthesis, were responsible for the stimulatory effects of increased nutrient supply in M. aeruginosa. Increased nitrogen supply directly promoted cyanobacterial growth by inducing the overexpression of the cell division regulatory protein FtsZ. NtcA, that regulates gene transcription related to both nitrogen assimilation and microcystin synthesis, was overexpressed under the high nitrogen condition, which consequently induced overexpression of 2 microcystin synthetases (McyC and McyF) and promoted microcystin synthesis. Elevated nitrogen supply induced the overexpression of proteins involved in gas vesicle organization (GvpC and GvpW), which may increase the buoyancy of M. aeruginosa. Increased phosphorus level indirectly affected growth and the synthesis of cellular substances in M. aeruginosa through the mediation of differentially expressed proteins related to carbon and phosphorus metabolism. This study provides a comprehensive description of changes in the proteome of M. aeruginosa in response to an increased supply of 2 key nutrients.

Sign in / Sign up

Export Citation Format

Share Document