Exceptional solvent tolerance in Yarrowia lipolytica is enhanced by sterols

Walker, Caleb; Ryu, Seunghyun; Trinh, Cong T.

doi:10.1016/j.ymben.2019.03.003

Cited by 54 publications

(47 citation statements)

References 80 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Reverse engineering of cds1 and cho1 was applied to enhance salt stress tolerance, consistent with the demonstration using whole‐genome sequencing in an evolutionary strain that mutation of pyruvate kinase is essential for restoring cell growth and that reintroduction of pyruvate kinase contributes to cell growth (Yu et al, ). Different from traditional metabolic engineering, reverse engineering can quickly locate the key target via metabolic model prediction (Lu et al, ) and multiple omics approaches (Walker et al, ), rather than extensive screening of metabolic pathway genes. Furthermore, membrane‐lipid‐associated genes reported to influence stress tolerance include cis‐trans isomerase Cti (Tan et al, ), PS synthase PssA (Tan et al, ), and desaturase Ole1 (Degreif et al, ).…”

Section: Discussionmentioning

confidence: 99%

“…In this study, redistributed membrane phospholipid driven by the module assembly of cds1 and cho1 contributed to the increase in membrane potential and membrane integrity, which significantly enhanced salt stress tolerance. Compared with long mutagenesis breeding (Cui et al, ), adaptive laboratory evolution (Walker et al, ), transporter engineering (Kell et al, ), this strategy is a more convenient and efficient way to achieve microbial tolerance. However, the rearranged phospholipid bilayer structure in strain Y03 is unknown, which may be important for exploring increased membrane potential and membrane integrity.…”

Section: Discussionmentioning

confidence: 99%

“…To increase the tolerance of industrial strains to environmental stress (Chakraborty, Winardhi, Morgan, Yan, & Kenney, ; Olin‐Sandoval et al, ), a series of strategies have been developed. These include global transcription machinery engineering (Alper, Moxley, Nevoigt, Fink, & Stephanopoulos, ), adaptive laboratory evolution (Walker, Ryu, & Trinh, ), mutagenesis breeding (Cui et al, ), transporter engineering (Kell, Swainston, Pir, & Oliver, ), and membrane engineering (Tan et al, ).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Engineering of membrane phospholipid component enhances salt stress tolerance in Saccharomyces cerevisiae

Yin

Zhu

Luo

et al. 2020

Biotech & Bioengineering

View full text Add to dashboard Cite

To increase the growth of industrial strains under environmental stress, the Saccharomyces cerevisiae BY4741 salt-tolerant strain Y00 that tolerates 1.2 M NaCl was cultured through nitroguanidine mutagenesis. The metabolomics and transcription data of Y00 were compared with those of the wild-type strain BY4741. The comparison identified two genes related to salt stress tolerance, cds1 and cho1. Modular assembly of cds1 and cho1 redistributed the membrane phospholipid component and decreased the ratio of anionic-to-zwitterionic phospholipid in strain Y03 that showed the highest salt tolerance. Therefore, significantly increased membrane potential and membrane integrity helped strain Y03 to resist salt stress (1.2 M NaCl). This study provides an effective membrane engineering strategy to enhance salt stress tolerance.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Engineering of membrane phospholipid component enhances salt stress tolerance in Saccharomyces cerevisiae

Yin

Zhu

Luo

et al. 2020

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…For example, through the expression of cis-trans isomerase (Cti) from Pseudomonas aeruginosa, the tufa was incorporated into the E. coli membrane, decreasing membrane fluidity, as a result, robustness and the bio-renewable fuel titer were improved (13). The content and composition of sterols can be changed by engineering the key enzymes associated with sterol biosynthesis or by changing the transcription level of the sterol biosynthesis enzymes, which are affected by global transcription factors, such as Upc2 and Ecm22 (14). For example, the expression of a key sterol C-5 desaturase FvC5SD from an edible mushroom in fission yeast improved the contents of ergosterol and oleic acid, which resulted in enhanced tolerance to ethanol and high temperature (15).…”

Section: Introductionmentioning

confidence: 99%

Enhancement of Sphingolipid Synthesis Improves Osmotic Tolerance of Saccharomyces cerevisiae

Zhu

Yin

Luo

et al. 2020

Appl Environ Microbiol

View full text Add to dashboard Cite

To enhance the growth performance of Saccharomyces cerevisiae under osmotic stress, mutant XCG001, which tolerates up to 1.5 M NaCl, was isolated through adaptive laboratory evolution (ALE). Comparisons of the transcriptome data of mutant XCG001 and the wild-type strain identified ELO2 as being associated with osmotic tolerance. In the ELO2 overexpression strain (XCG010), the contents of inositol phosphorylceramide (IPC; t18:0/26:0), mannosylinositol phosphorylceramide [MIPC; t18:0/22:0(2OH)], MIPC (d18:0/22:0), MIPC (d20:0/24:0), mannosyldiinositol phosphorylceramide [M(IP)2C; d20:0/26:0], M(IP)2C [t18:0/26:0(2OH)], and M(IP)2C [d20:0/26:0(2OH)] increased by 88.3 times, 167 times, 63.3 times, 23.9 times, 27.9 times, 114 times, and 208 times at 1.0 M NaCl, respectively, compared with the corresponding values of the control strain XCG002. As a result, the membrane integrity, cell growth, and cell survival rate of strain XCG010 increased by 24.4% ± 1.0%, 21.9% ± 1.5%, and 22.1% ± 1.1% at 1.0 M NaCl, respectively, compared with the corresponding values of the control strain XCG002 (wild-type strain with a control plasmid). These findings provided a novel strategy for engineering complex sphingolipids to enhance osmotic tolerance. IMPORTANCE This study demonstrated a novel strategy for the manipulation of membrane complex sphingolipids to enhance S. cerevisiae tolerance to osmotic stress. Elo2, a sphingolipid acyl chain elongase, was related to osmotic tolerance through transcriptome analysis of the wild-type strain and an osmosis-tolerant strain generated from ALE. Overexpression of ELO2 increased the content of complex sphingolipid with longer acyl chain; thus, membrane integrity and osmotic tolerance improved.

show abstract

“…While mammals require nutritional supplementation of thiamine from dietary sources, most bacteria, fungi, and plants can synthesize thiamine endogenously (20). One of the exceptions is the thiamine-auxotrophic oleaginous yeast Yarrowia lipolytica, which has recently emerged as an important industrial microbe with broad biotechnological applications due to its generally regarded as safe (GRAS) status (21), metabolic capability (22)(23)(24)(25)(26), and robustness (27)(28)(29). Hence, Y. lipolytica is an ideal eukaryotic host to study the fundamental effects of thiamine deficiency on cellular health.…”

mentioning

confidence: 99%

Understanding and Eliminating the Detrimental Effect of Thiamine Deficiency on the Oleaginous Yeast Yarrowia lipolytica

Walker

Ryu

Giannone

et al. 2020

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

Thiamine is a vitamin that functions as a cofactor for key enzymes in carbon and energy metabolism in all living cells. While most plants, fungi, and bacteria can synthesize thiamine de novo, the oleaginous yeast Yarrowia lipolytica cannot. In this study, we used proteomics together with physiological characterization to elucidate key metabolic processes influenced and regulated by thiamine availability and to identify the genetic basis of thiamine auxotrophy in Y. lipolytica. Specifically, we found that thiamine depletion results in decreased protein abundance for the lipid biosynthesis pathway and energy metabolism (i.e., ATP synthase), leading to the negligible growth and poor sugar assimilation observed in our study. Using comparative genomics, we identified the missing 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate synthase (THI13) gene for the de novo thiamine biosynthesis in Y. lipolytica and discovered an exceptional promoter, P3, that exhibits strong activation and tight repression by low and high thiamine concentrations, respectively. Capitalizing on the strength of our thiamine-regulated promoter (P3) to express the missing gene from Saccharomyces cerevisiae (scTHI13), we engineered a thiamine-prototrophic Y. lipolytica strain. By comparing this engineered strain to the wild-type strain, we revealed the tight relationship between thiamine availability and lipid biosynthesis and demonstrated enhanced lipid production with thiamine supplementation in the engineered thiamine-prototrophic Y. lipolytica strain. IMPORTANCE Thiamine plays a crucial role as an essential cofactor for enzymes involved in carbon and energy metabolism in all living cells. Thiamine deficiency has detrimental consequences for cellular health. Yarrowia lipolytica, a nonconventional oleaginous yeast with broad biotechnological applications, is a native thiamine auxotroph whose affected cellular metabolism is not well understood. Therefore, Y. lipolytica is an ideal eukaryotic host for the study of thiamine metabolism, especially because mammalian cells are also thiamine auxotrophic and thiamine deficiency is implicated in several human diseases. This study elucidates the fundamental effects of thiamine deficiency on cellular metabolism in Y. lipolytica and identifies genes and novel thiamine-regulated elements that eliminate thiamine auxotrophy in Y. lipolytica. Furthermore, the discovery of thiamine-regulated elements enables the development of thiamine biosensors with useful applications in synthetic biology and metabolic engineering.

show abstract

Exceptional solvent tolerance in Yarrowia lipolytica is enhanced by sterols

Cited by 54 publications

References 80 publications

Engineering of membrane phospholipid component enhances salt stress tolerance in Saccharomyces cerevisiae

Engineering of membrane phospholipid component enhances salt stress tolerance in Saccharomyces cerevisiae

Enhancement of Sphingolipid Synthesis Improves Osmotic Tolerance of Saccharomyces cerevisiae

Understanding and Eliminating the Detrimental Effect of Thiamine Deficiency on the Oleaginous Yeast Yarrowia lipolytica

Contact Info

Product

Resources

About