Large scale purification of an almond oleosin using an organic solvent procedure

Beisson, Frédéric; Ferté, Natalie; Voultoury, Robert; Arondel, Vincent

doi:10.1016/s0981-9428(01)01275-x

Cited by 40 publications

(32 citation statements)

References 26 publications

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“…Recombinant protein was found in inclusion bodies, which were isolated using standard extraction techniques and repeatedly washed with a high pH buffer to remove bound DNA from the positively charged hydrophilic arms. Protein was extracted using monophasic mixtures of sodium carbonate (pH 11), chloroform, and methanol as previously reported (27,28). Decreasing the size of the hydrophobic core to P ¼ 65 amino acids from P ¼ 87 amino acids increased protein expression 10-fold, and we obtained between 35-100 mg purified protein per liter of culture, with yield increasing when N and C were longer.…”

Section: Resultsmentioning

confidence: 65%

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Self-assembly of tunable protein suprastructures from recombinant oleosin

Vargo

Parthasarathy

Hammer

2012

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

102

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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Section: Resultsmentioning

confidence: 65%

“…The resulting inclusion body pellet was washed three times with 10 mL 200 mM Na 2 CO 3 (pH 11). Oleosin was extracted from the inclusion bodies using an organic solvent mixture (27,28). The pellet was resuspended in 10 mL 200 mM Na 2 CO 3 (pH 11).…”

Section: Methodsmentioning

confidence: 99%

Self-assembly of tunable protein suprastructures from recombinant oleosin

Vargo

Parthasarathy

Hammer

2012

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

102

130

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“…A certain degree of flocculation in protein bodies can be observed, which can be due to their hydrophobic character. Most of the almond proteins belong to the oleosin family with low-molecular-weight and poor water solubility, due to a long highly hydrophobic domain of about 70 amino acid residues (Beisson, Fert e, Voultoury, & Arondel, 2001). In some cases, protein bodies appear adsorbed on the oil droplet surface, forming bridges between them.…”

Section: Protein Denaturationmentioning

confidence: 99%

Effect of high pressure homogenisation and heat treatment on physical properties and stability of almond and hazelnut milks

Bernat

Cháfer

Rodríguez-García

et al. 2015

LWT - Food Science and Technology

137

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ElsevierBernat Pérez, N.; Cháfer Nácher, MT.; Rodriguez Garcia, J.; Chiralt A.; González Martínez, MC. (2015). Effect of high pressure homogenization and heat treatment on physical properties and stability of almond and hazelnut milks. Food Science and Technology. 62:488-496. doi:10.1016/j.lwt.2014.10.045. Effect of high pressure homogenisation and heat treatment on physical 1 properties and stability of almond and hazelnut milks (1) Institute of Food Engineering for the Development. 5(2) Food Technology Department. The effect of high pressure homogenisation (HPH) and heat treatments on physicochemical 12properties and physical stability of almond and hazelnut milks was studied. Vegetable milks were 13 obtained and homogenised by applying 62, 103 and 172 MPa (MF1, MF2 and MF3, respectively). 14 Untreated and MF3 samples were also submitted to two different heat treatments (85ºC/30 min 15 (LH) or 121ºC/15 min (HH)). Physical and structural properties of the products were greatly 16 affected by heat treatments and HPH. In almond milk, homogenised samples showed a significant 17 reduction in particle size, which turned from bimodal and polydisperse to monodisperse 18 distributions. Particle surface charge, clarity and Whiteness Index were increased and physical 19 stability of samples was improved, without affecting either viscosity or protein stability. Hazelnut 20 beverages showed similar trends, but HPH notably increased their viscosity while change their 21 rheological behaviour, which suggested changes in protein conformation. HH treatments caused 22 an increment of particle size due to the formation oil droplet-protein body clusters, associated 23 with protein denaturation. Samples submitted to the combined treatment MF3 and LH showed the 24 greatest stability. 25 26

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“…1A). A certain degree of protein flocculation was observed due to their hydrophobic character, since most almond proteins belong to the oleosin family, with low-molecular weight and poor water solubility (Beisson, Ferte, Voultoury, & Arondel, 2001). This low water-IJFS October 2015 Volume 4 pages 201-211 affinity of proteins contributes to the poor stability of the obtained emulsions.…”

Section: Sensory Analysismentioning

confidence: 99%

Probiotic fermented almond “milk” as an alternative to cow-milk yoghurt

Bernat

Cháfer

Chiralt

2015

IJFS

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Probiotics in almond-based matrices were considered as a means of obtaining fermented products which would cover both the current demand for health-promoting foods and for alternatives to standard yoghurts. Firstly, the combined effect of high pressure homogenisation (HPH) and heat treatment on the physical stability of almond "milk" was studied. The beverage was homogenised by applying 62, 103 and 172 MPa (MF1, MF2 and MF3 respectively); MF3 was also combined with two different heat treatments (85°C-30 min (LH) and 121°C-15 min (HH)). Both microstructure and colloidal stability were analysed in all the processed samples to select the most suitable treatment with which to obtain a stable product. The selected almond milk was then fermented with probiotic Lactobacillus reuteri and Streptococcus thermophilus and the final product was characterised throughout cold storage time (28 days) as to pH, acidity, serum retention and starter viability. A sensory evaluation and probiotic survival to in vitro digestion was also conducted. The results showed that the physical and structural almond-milk properties were affected by both HPH and heat treatments, obtaining the greatest stability in MF3-LH samples. The fermented milk permitted probiotic survivals above the level suggested as minimum for ensuring health benefits during the entire controlled time and, hence, can be considered as a functional food. No differences in the sensory acceptability of the product were found between 1 and 28 storage days. Therefore, a new, functional, fermented product was developed, which was suitable for targeted groups, such as the lactose-intolerant and cow-milk-protein allergic populations.

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Large scale purification of an almond oleosin using an organic solvent procedure

Cited by 40 publications

References 26 publications

Self-assembly of tunable protein suprastructures from recombinant oleosin

Self-assembly of tunable protein suprastructures from recombinant oleosin

Effect of high pressure homogenisation and heat treatment on physical properties and stability of almond and hazelnut milks

Probiotic fermented almond “milk” as an alternative to cow-milk yoghurt

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