scholarly journals What You Extract Is What You Get: Different Methods of Protein Extraction from Hemp Seeds

Separations ◽  
2021 ◽  
Vol 8 (12) ◽  
pp. 231
Author(s):  
Annalisa Givonetti ◽  
Chiara Cattaneo ◽  
Maria Cavaletto

Cannabis sativa L. seeds are rich in essential polyunsaturated fatty acids and highly digestible proteins, with a good nutritional value. Proteomics studies on hempseed reported so far have mainly been conducted on processed seeds and, to our knowledge, no optimization of protein extraction from hemp seeds has been performed. This study investigates the SDS-PAGE profile of hempseed proteins comparing different methods of extraction, (Osborne sequential extraction, TCA/acetone, MTBE/methanol, direct protein solubilization of defatted hempseed flour), two conditions to keep low temperature during seed grinding (liquid nitrogen or ice) and two solubilization buffers (urea-based or Laemmli buffer). Among the tested conditions, the combination of liquid nitrogen + TCA/acetone + Laemmli buffer was not compatible with SDS-PAGE of proteins. On the other hand, urea-based buffer achieved more reproducible results if combined with all the other conditions. TCA/acetone, MTBE/methanol, and direct protein solubilization of defatted hempseed flour demonstrated a good overview of protein content, but less abundant proteins were poorly represented. The Osborne sequential separation was helpful in diluting abundant proteins thus enhancing the method sensitivity.

Foods ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1793
Author(s):  
Marta Barral-Martinez ◽  
Maria Fraga-Corral ◽  
Pascual Garcia-Perez ◽  
Jesus Simal-Gandara ◽  
Miguel A. Prieto

The search for waste minimization and the valorization of by-products are key practices for good management and improved sustainability in the food industry. The production of almonds generates a large amount of waste, most of which is not used. Until now, almonds have been used for their high nutritional value as food, especially almond meat. The other remaining parts (skin, shell, hulls, etc.) are still little explored, even though they have been used as fuel by burning or as livestock feed. However, interest in these by-products has been increasing as they possess beneficial properties (caused mainly by polyphenols and unsaturated fatty acids) and can be used as new ingredients for the food, cosmetic, and pharmaceutical industries. Therefore, it is important to explore almond’s valorization of by-products for the development of new added-value products that would contribute to the reduction of environmental impact and an improvement in the sustainability and competitiveness of the almond industry.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 454b-454
Author(s):  
Yiwei Jiang ◽  
Chon C. Lim ◽  
Rajeev Arora

Onion (Allium cepa L. cv Sweet Sandwich) and Chrysanthemum (Dendranthema grandiflora `Sunny Denise') tissues were used to investigate protein changes associated with recovery from freeze–thaw injury. Medium-sized onions were slowly frozen to either –4 or –9 °C, subsequently thawed, and divided in two halves. One half was used immediately for ion leakage (IL) measurements and total and microsomal protein extraction, whereas the other half was allowed to recover at 6 to 8 °C in the dark for 4 to 5 days. Chrysanthemum leaves were frozen to –3.75 °C, and allowed to recover first at 6 to 8 °C in the dark (1 d) and then under 12-h photoperiod at 18 °C (3–4 d). Results indicate a 1.4- and 2.5-fold higher IL, compared to control, from onion tissues frozen to –4 or –9 °C, respectively. IL in –4 °C-treated tissues was the same as respective control following recovery; however, it was further enhanced to 3.6-fold in –9 °C-treated samples. Chrysanthemum leaf tissue exhibited a 1.6-fold increase in ion leakage following injury, but completely recovered to control levels after 4 to 5 d. SDS-PAGE profiles revealed an absence of a 25-kDa microsomal protein in the injured onion tissues but, its up regulation during recovery only in reversibly injured tissues. Data also indicated an accumulation of 36-kDa soluble protein in chrysanthemum leaf tissues during recovery. Experiments are underway to further characterize these protein changes.


Author(s):  
T. Inoué ◽  
H. Koike

Low temperature scanning electron microscopy (LTSEM) is useful to avoid artifacts such as deformation and extraction, because specimens are not subjected to chemical fixation, dehydration and critical-point drying. Since Echlin et al developed a LTSEM, many techniques and instruments have been reported for observing frozen materials. However, intracellular structures such as mitochondria and endoplasmic reticulum have been unobservable by the method because of the low resolving power and inadequate specimen preparation methods. Recently, we developed a low temperature SEM that attained high resolutions. In this study, we introduce highly magnified images obtained by the newly developed LTSEM, especially intracellular structures which have been rapidly frozen without chemical fixation.[Specimen preparations] Mouse pancreas and brown adipose tissues (BAT) were used as materials. After the tissues were removed and cut into small pieces, the specimen was placed on a cryo-tip and rapidly frozen in liquid propane using a rapid freezing apparatus (Eiko Engineering Co. Ltd., Japan). After the tips were mounted on the specimen stage of a precooled cryo-holder, the surface of the specimen was manually fractured by a razor blade in liquid nitrogen. The cryo-holder was then inserted into the specimen chamber of the SEM (ISI DS-130), and specimens were observed at the accelerating voltages of 5-8 kV. At first the surface was slightly covered with frost, but intracellular structures were gradually revealed as the frost began to sublimate. Gold was then coated on the specimen surface while tilting the holder at 45-90°. The holder was connected to a liquid nitrogen reservoir by means of a copper braid to maintain low temperature.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hitomi Nakamura ◽  
Moeka Yoshikawa ◽  
Naoko Oda-Ueda ◽  
Tadashi Ueda ◽  
Takatoshi Ohkuri

AbstractGenerally, intermolecular disulfide bond contribute to the conformational protein stability. To identify sites where intermolecular disulfide bond can be introduced into the Fab’s constant domain of the therapeutic IgG, Fab mutants were predicted using the MOE software, a molecular simulator, and expressed in Pichia pastoris. SDS-PAGE analysis of the prepared Fab mutants from P. pastoris indicated that among the nine analyzed Fab mutants, the F130C(H):Q124C(L), F174C(H):S176C(L), V177C(H):Q160C(L), F174C(H):S162C(L), F130C(H):S121C(L), and A145C(H):F116C(L) mutants mostly formed intermolecular disulfide bond. All these mutants showed increased thermal stability compared to that of Fab without intermolecular disulfide bond. In the other mutants, the intermolecular disulfide bond could not be completely formed, and the L132C(H):F118C(L) mutant showed only a slight decrease in binding activity and β-helix content, owing to the exertion of adverse intermolecular disulfide bond effects. Thus, our comprehensive analysis reveals that the introduction of intermolecular disulfide bond in the Fab’s constant domain is possible at various locations. These findings provide important insights for accomplishing human Fab stabilization.


2021 ◽  
Author(s):  
Xiaodan Lu ◽  
Rongbin Zhong ◽  
Ling Hu ◽  
Luyao Huang ◽  
Lijiao Chen ◽  
...  

Abstract Large yellow croaker roe phospholipids (LYCRPLs) has great nutritional value because of containing rich docosahexaenoic acid (DHA), which is a kind of n-3 polyunsaturated fatty acids (n-3 PUFAs). In...


Antioxidants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 538
Author(s):  
Vita Maria Marino ◽  
Teresa Rapisarda ◽  
Margherita Caccamo ◽  
Bernardo Valenti ◽  
Alessandro Priolo ◽  
...  

Hazelnut peel (HNP), a by-product from the chocolate industry, is considered to be a suitable ingredient to be included in the diet of ruminants. This study aimed to evaluate the effect of feeding dairy ewes with a diet containing HNP on ripened cheese quality, including fatty acid (FA) profile, cholesterol, and tocopherol content, as well as stability during storage under commercial conditions. In total, 10 experimental cheeses were produced with bulk milk obtained from ewes fed a commercial concentrate (C group; n = 5) or a concentrate containing 36% HNP in dry matter (HNP group; n = 5). After 40 days of aging, each cheese was sub-sampled into three slices: one was analyzed immediately (C0 and HNP0), and the other two were refrigerated and analyzed after seven days (C7 and HNP7) and 14 days (C14 and HNP14), respectively. Compared to C, HNP cheese had more than twice as many tocopherols and mono-unsaturated FA and respectively 38% and 24% less of cholesterol and saturated FA. Tocopherols and cholesterol levels remained rather stable up to 14 days of storage regardless of the experimental group, suggesting no cholesterol oxidation. Therefore, the inclusion of HNP in ewe diets could be a valid resource to produce cheese with a healthier lipid profile and higher tocopherols content.


Animals ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 59
Author(s):  
Natalia Śmietana ◽  
Remigiusz Panicz ◽  
Małgorzata Sobczak ◽  
Przemysław Śmietana ◽  
Arkadiusz Nędzarek

The aim of the study was to present a comprehensive characterisation of crayfish meat, which is crucial to assess its potential usefulness in the food industry. To this end, we assessed the yield, basic chemical composition (protein, fat, minerals), nutritional value (amino acid and fatty acid profiles, essential amino acid index (EAAI), chemical score of essential amino acids (CS), hypocholesterolaemic/hypercholesterolaemic ratio (h/H), atherogenicity (AI) and thrombogenicity (TI) indices), as well as culinary value (lab colour, texture, sensory characteristics, structure) of the meat of spiny-cheek crayfish (Faxonius limosus) (n = 226) from Lake Sominko (Poland) harvested in May–September 2017. Crayfish meat, especially that from the abdomen, was shown to have high nutritional parameters. It is lean (0.26% of fat), with a favourable fatty acid profile and a very high quality of fat (PUFA (sum of polyunsaturated fatty acids):SFA (sum of saturated fatty acids), n-6/n-3, h/H, AI, TI) and protein (high CS and EAAI). It is also a better source of Ca, K, Mg, Na, P, and Cu than meat from slaughter animals. Hence, crayfish meat can be an alternative to livestock meat in the human diet. Owing to its culinary value (delicateness, weak game flavour, and odour), it meets the requirements of the most demanding consumers, i.e., children and older people.


2021 ◽  
Vol 22 (4) ◽  
pp. 1554
Author(s):  
Tawhidur Rahman ◽  
Mingxuan Shao ◽  
Shankar Pahari ◽  
Prakash Venglat ◽  
Raju Soolanayakanahally ◽  
...  

Cuticular waxes are a mixture of hydrophobic very-long-chain fatty acids and their derivatives accumulated in the plant cuticle. Most studies define the role of cuticular wax largely based on reducing nonstomatal water loss. The present study investigated the role of cuticular wax in reducing both low-temperature and dehydration stress in plants using Arabidopsis thaliana mutants and transgenic genotypes altered in the formation of cuticular wax. cer3-6, a known Arabidopsis wax-deficient mutant (with distinct reduction in aldehydes, n-alkanes, secondary n-alcohols, and ketones compared to wild type (WT)), was most sensitive to water loss, while dewax, a known wax overproducer (greater alkanes and ketones compared to WT), was more resistant to dehydration compared to WT. Furthermore, cold-acclimated cer3-6 froze at warmer temperatures, while cold-acclimated dewax displayed freezing exotherms at colder temperatures compared to WT. Gas Chromatography-Mass Spectroscopy (GC-MS) analysis identified a characteristic decrease in the accumulation of certain waxes (e.g., alkanes, alcohols) in Arabidopsis cuticles under cold acclimation, which was additionally reduced in cer3-6. Conversely, the dewax mutant showed a greater ability to accumulate waxes under cold acclimation. Fourier Transform Infrared Spectroscopy (FTIR) also supported observations in cuticular wax deposition under cold acclimation. Our data indicate cuticular alkane waxes along with alcohols and fatty acids can facilitate avoidance of both ice formation and leaf water loss under dehydration stress and are promising genetic targets of interest.


2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1038-1038
Author(s):  
Michael Miklus ◽  
Pedro Prieto ◽  
Cynthia Barber ◽  
Robert Rhoads ◽  
Samer El-Kadi

Abstract Objectives The objectives of this study were to determine the effect of 2’fucosyllactose (2’FL) and fat blends on growth, body composition and fatty acid profile of the liver and brain using the neonatal pig as a model for the human infant. Methods Pigs (3 d old) were randomly assigned to either: 1. control, 2. Palm Olein (PO) fat blend – Low 2'-FL, 3. PO – High 2'-FL, 4. High oleic acid (HO) – Low 2'-FL, 5. HO FB – High 2'-FL, 6. PO FB – GLA, or 7. kept with their sows. Pigs in groups 1 to 6 received 250 ml·kg−1·d−1 of formula in 5 equal meals for 15 d. On day 14 of the study, groups 1–6 received intraperitoneal E. coli LPS challenge at 100 µg·kg−1 weight. Results Body weight was greater for piglets fed by sows than those in the other groups (P < 0.001). In addition, % fat and bone mineral content were higher in the sow-fed group while lean % was less sow-fed piglets (group 7) compared with those in the other groups (P < 0.05). Only longissimus weight expressed as a % of body weight, was greater for group 7 compared with all other groups (P < 0.001). Soleus, semitendinosus, brain, heart and spleen weights as a % of body weight were similar across all groups. However, liver weight as a % of body weight was greater in groups 1–6 (3.7%) compared with group 7 (2.8%; P < 0.001). The proportion of brain 16:1 fatty acid was less (0.83%) for groups 1–6 than for group 7 pigs (1.08%; P < 0.0001). The proportion of 20:3 N6 was greatest (0.66%) for group 3 compared with groups 1 and 4 (0.55%; P < 0.05). In addition, the proportion of 20:5 N3 was greatest (0.12%) for group 3 compared with groups 1 and 7 (0.07%; P < 0.05). The proportion of liver 16:1, 18:0, and 18:1 cis-11 fatty acids were greater for group 7 (2.3, 23, 2.2%) than groups 1–6 (0.2, 20, 1.2%; P < 0.0001). Conversely, the contribution of 14:0, 18:1 cis-9, 18:3 N6 cis-6,9,12, and 22:6 N3 were greater for pigs in groups 1–6 (1.3, 0.6, and 14, 7.8%) compared with those in group 7 (0.5, 8.5, 0.2 and 3.5%; P < 0.0001). Conclusions Our data suggest that feeding 2’fucosyllactose had no effect on the body weight gain and composition in neonatal pigs. Our data also suggest that dietary fatty acids have a greater effect on liver than on brain fatty acid composition. Funding Sources Funding for the work was provided by Perrigo Nutritionals, LLC.


2003 ◽  
Vol 56 (9) ◽  
pp. 903 ◽  
Author(s):  
Gavin E. Collis ◽  
Dieter Wege

Addition of 2-diazopropane to 1,4-naphthoquinone at low temperature, followed by in situ enolization and acetylation or silylation gave 3,3-dimethyl-1H-benz[f]indazol-4,9-diyl diacetate and 3,3-dimethyl-9-(t-butyl-dimethylsilyloxy)-1H-benz[f]indazol-4-ol, respectively. Functional group manipulation of the latter compound provided a number of other 4,9-disubstituted 3,3-dimethyl-3H-benz[f]indazoles. Irradiation of the diacetate led to clean extrusion of nitrogen to give the naphtho[b]cycloproparene and an alkene. Attempts to elaborate the cycloproparene into the derived cyclopropanaphthoquinone were unsuccessful. Of the other 4,9-disubstituted 3,3-dimethyl-3H-benz[f]indazoles examined, only the compound possessing an acetoxy group at C9 was photoactive, and afforded the expected cycloproparene and alkene. Compounds bearing a hydroxy or alkoxy group at C9 were photochemically inert.


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