Cyanobacteria Total Lipid Extraction from Polycarbonate Filters v1

This protocol is designed/used for extraction of total cellular lipids from cyanobacteria samples (either lab cultures or field samples) collected on polycarbonate filters for use in lipid analysis and quantification via mass spectrometry. Please contact Dr. Steven Wilhelm ([email protected]) or Robbie M. Martin ([email protected]) for additional information regarding this protocol. Modified from Guan, X. L., Riezman, I., Wenk, M. R., & Riezman, H. (2010). Yeast lipid analysis and quantification by mass spectrometry. Methods in Enzymology, 470, 369-391.

Seipin and Nem1 establish discrete ER subdomains to initiate yeast lipid droplet biogenesis

Lipid droplets (LDs) are fat storage organelles that originate from the endoplasmic reticulum (ER). Relatively little is known about how sites of LD formation are selected and which proteins/lipids are necessary for the process. Here, we show that LDs induced by the yeast triacylglycerol (TAG)-synthases Lro1 and Dga1 are formed at discrete ER subdomains defined by seipin (Fld1), and a regulator of diacylglycerol (DAG) production, Nem1. Fld1 and Nem1 colocalize to ER–LD contact sites. We find that Fld1 and Nem1 localize to ER subdomains independently of each other and of LDs, but both are required for the subdomains to recruit the TAG-synthases and additional LD biogenesis factors: Yft2, Pex30, Pet10, and Erg6. These subdomains become enriched in DAG. We conclude that Fld1 and Nem1 are both necessary to recruit proteins to ER subdomains where LD biogenesis occurs.

Seipin and Nem1 establish discrete ER subdomains to initiate yeast lipid droplet biogenesis

ABSTRACTLipid droplets (LDs) are fat storage organelles that originate from the endoplasmic reticulum (ER). Relatively little is known about how sites of LD formation are selected, and which proteins/lipids are necessary for the process. Here, we show that LDs induced by the yeast triacylglycerol (TAG)-synthases Lro1 and Dga1 are formed at discrete ER subdomains defined by seipin (Fld1), and a regulator of diacylglycerol (DAG) production, Nem1. Fld1 and Nem1 colocalize to ER-LD contact sites. We find that Fld1 and Nem1 localize to ER subdomains independently of each other and of LDs, but both are required for the subdomains to recruit the TAG synthases and additional LD biogeneiss factors: Yft2, Pex30, Pet10, and Erg6. These subdomains become enriched in DAG. We conclude that Fld1 and Nem1 are both necessary to recruit proteins to ER subdomains where LD biogenesis occurs.

Yeast lipid-based biofuels and oleochemicals from lignocellulosic biomass: life cycle impact assessment

Over the last few decades, much attention has been paid to various applications of microbial oil, including for transportation.