A system for purification of recombinant proteins in Escherichia coli via artificial oil bodies constituted with their oleosin-fused polypeptides

Peng, Ching‐I; Chen, Jeff C.F; Shyu, Douglas J. H.; Chen, Ming-Jinn; Tzen, Jason T.C.

doi:10.1016/j.jbiotec.2004.03.013

Cited by 49 publications

(40 citation statements)

References 27 publications

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“…The sizes of artificial oil bodies could be controlled by changing the ratio of triacylglycerol over oil-body protein, whereas both thermostability and structural stability of artificial oil bodies decreased as their size increased, and vice versa [87]. For encapsulation of artificial oil bodies, recombinant oleosins expressed in Escherichia coli were found comparable to native oleosins isolated from seed oil bodies [141,142]. It has been demonstrated that artificial oil bodies could be stabilized by oleosin or caleosin, but not steroleosin, and the average sizes (50-200 nm) of artificial oil bodies constituted with caleosin were 10-times smaller than those (0.5-2 μm) constituted with oleosin ( Figure 5) [143].…”

Section: Artificial Oil Bodies In Basic Research Studiesmentioning

confidence: 99%

Integral Proteins in Plant Oil Bodies

Tzen

2012

ISRN Botany

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Hydrophobic storage neutral lipids are stably preserved in specialized organelles termed oil bodies in the aqueous cytosolic compartment of plant cells via encapsulation with surfactant molecules including phospholipids and integral proteins. To date, three classes of integral proteins, termed oleosin, caleosin, and steroleosin, have been identified in oil bodies of angiosperm seeds. Proposed structures, targeting traffic routes, and biological functions of these three integral oil-body proteins were summarized and discussed. In the viewpoint of evolution, isoforms of oleosin and caleosin are found in oil bodies of pollens as well as those of more primitive species; moreover, caleosin-and steroleosin-like proteins are also present in other subcellular locations besides oil bodies. Technically, artificial oil bodies of structural stability similar to native ones were successfully constituted and seemed to serve as a useful tool for both basic research studies and biotechnological applications.

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Section: Artificial Oil Bodies In Basic Research Studiesmentioning

confidence: 99%

Integral Proteins in Plant Oil Bodies

Tzen

2012

ISRN Botany

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“…Whether this is the case for LDSP as well, or if it localizes to the LD merely due to its high hydrophobicity, remains to be determined. It should be pointed out that oleosins have the ability to stabilize artificial LDs (Peng et al, 2004), suggesting that they are capable of spontaneous insertion into a lipid matrix.…”

Section: Ldsp Of Nannochloropsismentioning

confidence: 99%

A Lipid Droplet Protein of Nannochloropsis with Functions Partially Analogous to Plant Oleosins

et al. 2012

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(A.V., C.B.); and Aurora Algae, Hayward, California 94545 (S.B.B., B.V.)As our understanding of the dynamics of lipid droplets (LDs) in animal, plant, and fungal cells is rapidly evolving, still little is known about the formation and turnover of these organelles in microalgae. Yet with the growing importance of algal feedstock for the production of biofuels and high-value lipids, there is a need to understand the mechanisms of LD dynamics in microalgae. Thus, we investigated the proteins associated with LDs of the emerging heterokont model alga Nannochloropsis sp. and discovered an abundant hydrophobic lipid droplet surface protein (LDSP) with unique primary sequence but structural similarities to other LD proteins. LDSP abundance in Nannochloropsis cells closely tracked the amount of triacylglycerols during conditions of oil accumulation and degradation. Functional characterization of LDSP in an Arabidopsis (Arabidopsis thaliana) OLEOSIN1-deficient mutant allowed a separation of its physical and structural properties in its interaction with LDs from its physiological or biochemical activities. Although LDSP presence in Arabidopsis predictably affected LD size, it could not reverse the physiological impact of OLEOSIN deficiency on triacylglycerol hydrolysis during germination.

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“…The two legs of the hairpin are helical, possibly forming a coiled-coil (17,20). Recently, oleosin has been shown to stabilize artificial oil bodies and emulsions (22)(23)(24)(25)(26). We postulated that the surfactant-like block architecture of oleosin would make it a logical starting point for the creation of tunable self-assembled protein suprastructures.…”

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confidence: 99%

Self-assembly of tunable protein suprastructures from recombinant oleosin

Vargo

Parthasarathy

Hammer

2012

Proc. Natl. Acad. Sci. U.S.A.

130

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Using recombinant amphiphilic proteins to self-assemble suprastructures would allow precise control over surfactant chemistry and the facile incorporation of biological functionality. We used cryo-TEM to confirm self-assembled structures from recombinantly produced mutants of the naturally occurring sunflower protein, oleosin. We studied the phase behavior of protein self-assembly as a function of solution ionic strength and protein hydrophilic fraction, observing nanometric fibers, sheets, and vesicles. Vesicle membrane thickness correlated with increasing hydrophilic fraction for a fixed hydrophobic domain length. The existence of a bilayer membrane was corroborated in giant vesicles through the localized encapsulation of hydrophobic Nile red and hydrophilic calcein. Circular dichroism revealed that changes in nanostructural morphology in this family of mutants was unrelated to changes in secondary structure. Ultimately, we envision the use of recombinant techniques to introduce novel functionality into these materials for biological applications.protein surfactants | self-assembled suprastructure | cryogenic transmission microscopy

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A system for purification of recombinant proteins in Escherichia coli via artificial oil bodies constituted with their oleosin-fused polypeptides

Cited by 49 publications

References 27 publications

Integral Proteins in Plant Oil Bodies

Integral Proteins in Plant Oil Bodies

A Lipid Droplet Protein of Nannochloropsis with Functions Partially Analogous to Plant Oleosins

Self-assembly of tunable protein suprastructures from recombinant oleosin

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