Journal: Microorganisms

Article Title: Citrate-Mediated Acyl-CoA Synthesis Is Required for the Promotion of Growth and Triacylglycerol Production in Oleaginous Yeast Lipomyces starkeyi

doi: 10.3390/microorganisms9081693

Figure Lengend Snippet: List of primers and primer sequences used for plasmid construction in this study.

Article Snippet: The DNA fragment used for yeast transformation was obtained by digesting the plasmid pKS-PTDH3-ACL1-sNAT1-PTDH3-ACL2 with ApaI (Takara Bio).

Techniques: Plasmid Preparation, Sequencing

Journal: Microorganisms

Article Title: Citrate-Mediated Acyl-CoA Synthesis Is Required for the Promotion of Growth and Triacylglycerol Production in Oleaginous Yeast Lipomyces starkeyi

doi: 10.3390/microorganisms9081693

Figure Lengend Snippet: Overview of the principal metabolic pathways for TAG synthesis in L. starkeyi. Abbreviations for genes and metabolites are indicated as follows: ACC1 , acetyl-CoA carboxylase; ACL1 / ACL2 , ATP-citrate lyase; ACS1 , acetyl-CoA synthetase; α-KG, α-ketoglutarate; DAG, diacylglycerol; DHAP, dihydroxyacetone phosphate; FAS1 / FAS2 , fatty acid synthase subunit; Fructose-1,6-BP, fructose-1,6-bisphosphatase; Glucose-6-P, glucose-6-phosphate; Glyceraldehyde-3-P, glyceraldehyde-3-phosphate; Glycerol-3-P, glycerol-3-phosphate; Lyso-PA, lysophosphatidic acid; NADPH, reduced nicotinamide adenine dinucleotide phosphate; OAA, oxaloacetate; PA, phosphatidic acid; Ribulose-5-P, ribulose-5-phosphate; TAG, triacylglycerol.

Article Snippet: The DNA fragment used for yeast transformation was obtained by digesting the plasmid pKS-PTDH3-ACL1-sNAT1-PTDH3-ACL2 with ApaI (Takara Bio).

Techniques:

Journal: Microorganisms

Article Title: Citrate-Mediated Acyl-CoA Synthesis Is Required for the Promotion of Growth and Triacylglycerol Production in Oleaginous Yeast Lipomyces starkeyi

doi: 10.3390/microorganisms9081693

Figure Lengend Snippet: The effect of ATP-citrate lyase deletion on TAG synthesis in an S medium containing glucose. ∆ lslig4 and ∆ lslig4 ∆ acl1 ∆ acl2 were cultured in an S medium containing glucose at 30 °C for five days in a small-scale culture. Symbols: closed circles, ∆ lslig4 ; closed squares, ∆ lslig4 ∆ acl1 ∆ acl2 ; gray bars, ∆ lslig4 ; black bars, ∆ lslig4 ∆ acl1 ∆ acl2 . N.D. indicates not detected. ( A ) Cell concentration (cells/mL), ( B ) glucose concentration (g/L), ( C ) TAG concentration (g/L), ( D ) amount of TAG per cell (mg/10 8 cells), and ( E ) relative expression levels of genes involved in acyl-CoA synthesis ( ACL1 , ACL2 , ACC1 , FAS1 , FAS2 , and ACS1 ). The transcription levels with relative quantitative method normalized by 18S rRNA. The data are indicated as the mean ± SEM of the three independent experiments. Asterisks indicate statistically significant differences between ∆ lslig4 and ∆ lslig4 ∆ acl1 ∆ acl2 ( t -test; * p < 0.05).

Article Snippet: The DNA fragment used for yeast transformation was obtained by digesting the plasmid pKS-PTDH3-ACL1-sNAT1-PTDH3-ACL2 with ApaI (Takara Bio).

Techniques: Cell Culture, Concentration Assay, Expressing

Journal: Microorganisms

Article Title: Citrate-Mediated Acyl-CoA Synthesis Is Required for the Promotion of Growth and Triacylglycerol Production in Oleaginous Yeast Lipomyces starkeyi

doi: 10.3390/microorganisms9081693

Figure Lengend Snippet: The effect of ATP-citrate lyase deletion on TAG synthesis on an S medium containing glucose and acetate. Symbols: closed circles, ∆ lslig4 ; closed squares, ∆ lslig4 ∆ acl1 ∆ acl2 ; gray bars, ∆ lslig4 ; black bars, ∆ lslig4 ∆ acl1 ∆ acl2 . N.D. indicates not detected. ( A – E ) The analysis of cell concentration, glucose concentration, TAG concentration, amount of TAG per cell, and relative expression levels of genes involved in acyl-CoA synthesis were performed as described in , except that the S medium containing glucose was replaced with an S medium containing glucose and acetate. Asterisks indicate statistically significant differences between ∆ lslig4 and ∆ lslig4 ∆ acl1 ∆ acl2 ( t -test; * p < 0.05).

Article Snippet: The DNA fragment used for yeast transformation was obtained by digesting the plasmid pKS-PTDH3-ACL1-sNAT1-PTDH3-ACL2 with ApaI (Takara Bio).

Techniques: Concentration Assay, Expressing