Field pennycress, Thlaspi arvense L., is a new oil seed cover crop being commercialized as a source of biofuel in the United States. Along the path to commercialization, pennycress seed and meal will ...be marketed as animal feed. As a member of the Brassicacaeae family, pennycress seed is abundant in glucosinolates which are anti-nutritional factors that make the seed and meal less palatable to animals. More specifically, sinigrin has been identified as the only glucosinolate present in pennycress seed. As work continues to develop varieties low in glucosinolates, a rapid assay is needed to determine the sinigrin level in pennycress seed.
The objective of this study was to develop and utilize a high throughput, rapid assay to screen pennycress seeds from wild populations for total sinigrin content. Pennycress seeds were ground in potassium phosphate buffer with endogenous myrosinase to catalyze the enzymatic hydrolysis of sinigrin to equimolar amounts of glucose. Glucose content was determined using a blood glucose meter and test strips. From the molar concentration of glucose, the molar concentration of sinigrin was known and the sinigrin level in seeds was calculated in µmol/g. The results were confirmed by HPLC/MS analysis.
A rapid and inexpensive method was developed and used to screen seeds of 429 wild pennycress lines for sinigrin. Sinigrin levels in seed ranged from 25 to 85 μmol/g in the pennycress lines screened. Environmental factors also seemed to contribute to the levels of sinigrin in seeds found within the line ‘tt8-t/ARV1’ grown at ten different locations in the Midwest, USA.
This rapid and inexpensive assay will allow breeders to efficiently screen lines and select for varieties that produce seeds with low sinigrin levels for improved feed value.
•Introduces new rapid inexpensive assay for detecting glucosinolates in pennycress seed.•Provides method development and validation for determining glucose levels in seed.•Screened 429 wild populations of pennycress for glucosinolate levels.•Evaluated glucosinolate levels across growing locations and years.
Pennycress protein isolate (PcPI) has inferior gelation and water holding properties compared to market leader soy protein, preventing its use in food applications such as meat analogues. Therefore, ...this work aimed to induce polymerization of PcPI by either cold atmospheric plasma (CAP) or transglutaminase (TG) to improve functionality and texturization potential. Micro-compounding was utilized to determine bench-scale texturization potential of unmodified, CAP, and TG modified PcPI hydrated at 50% moisture. CAP treatment following dielectric barrier discharge (DBD) induced polymerization primarily through disulfide linkages. Whereas, TG resulted in a relatively higher extent of polymerization induced by disulfide linkages and other covalent interactions involving mostly cruciferin acidic subunits. Compared to unmodified PcPI, the gel strength doubled and tripled post CAP and TG treatments, respectively. TG treatment caused a significant increase in water holding capacity by 20%. Unmodified PcPI did not form fibrous structures upon micro-compounding, instead it formed a soft mass with low resilience and cohesiveness. CAP modified PcPI (PcPI-CP) had the lowest water holding properties, which resulted in a relatively hard (significantly highest mechanical energy), dense fibrous structures. Due to high gelation strength and water holding capacity, TG modified PcPI (PcPI-TG) resulted in less dense fibrous structures with more air incorporation (significantly higher void %) relative to PcPI control and PcPI-CP. PcPI-TG texturized mass had significantly higher chewiness that is desirable for meat analogues. This research, for the first time, confirmed the potential success of induced polymerization of PcPI, especially with TG, in enhancing gelation and texturization potential.
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•Cold plasma (CP) and transglutaminase (TG) induced pennycress protein crosslinking.•Protein crosslinking by CP occurred through intermolecular disulfide linkages.•Protein crosslinking by TG occurred via intra- and inter-molecular covalent bonds.•Protein crosslinking contributed to enhanced gel strength and texturization.•TG modification resulted in superior properties of texturized pennycress protein.
Field pennycress (Thlaspi arvense L.) has been adapted as an off-season cover crop for the midwestern United States. It is a non-food crop that can be grown as a winter annual crop and harvested ...in-between corn (Zea mays L.) and soybean (Glycine max (L.) Merr.) seasons. High oleic pennycress (HOP) has been developed from field pennycress with the intention of producing pennycress oil with more desirable properties for industrial applications. In this work, the composition and physical properties of HOP oil have been characterized. As expected, the oil was predominantly composed of oleic acid (62.6%) and had a low saturated fatty acid content (4.1%). The cold flow properties of HOP oil were superb, with a cloud point of − 24 °C and a pour point of − 35 °C. Additional properties including antioxidant content, viscosity, oxidative stability, and lubricity were analyzed. Compared with field pennycress oil, HOP oil had better cold flow properties, but lower oxidative stability and antioxidant content. Thus, HOP oil has advantages in low temperature applications, while field pennycress oil is more suitable for high temperature applications. Structure property relationships between the fatty acid composition and physical properties of the oils are discussed.
•High oleic pennycress oil displays remarkable cold flow properties.•The high oleic variety exhibits moderate oxidative stability, excellent viscosity index, and adequate lubricity.•Compared to field pennycress oil, the high oleic variety has advantages in low temperature applications.
•Pennycress seed protein isolated by saline extraction and acid precipitation.•Protein isolates were sinigrin-free and had high purity (at least 90% protein).•Acid-precipitated protein isolate had ...superior amino acid quality.•Saline-extracted protein was highly soluble with excellent emulsifying properties.•Acid-precipitated protein had better foaming properties and heat stability.
This study evaluated two methods, saline extraction (SE) and conventional acid precipitation (AP), to recover proteins from pennycress (Thlaspi arvense L.) seed meal. SE was done using 0.1M NaCl at 50°C while AP involved alkaline extraction (pH 10) first followed by protein precipitation at pH 4. Composition, amino acid profiles, and functional properties (solubility, foaming, emulsification, water-holding capacity, heat coagulability) of the resultant protein extracts were compared. SE and AP produced pennycress protein extracts that were sinigrin-free and containing at least 90% (db) crude protein, which classifies the extracts as protein isolates (PI). Extraction method had major influence on the amino acid profiles and functional properties of the protein isolates. Pennycress SEPI was markedly more soluble (68–91% solubility at pH 2 and ≥7) and had excellent emulsifying properties that were clearly superior to those of APPI. On the other hand, APPI had better foaming properties and was more stable to heating than SEPI. These results strongly demonstrate that high-purity pennycress seed protein isolates can be produced by either saline extraction or acid precipitation and have functional properties that are desirable for non-food uses.
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•Fe3O4/graphene oxide-phenylalanine bisulfate core-shell catalyst was synthesized.•A higher biodiesel yield was prepared from field pennycress seed oil.•The catalyst is ...environmentally friendly, reusable and easily recovered.•The developed technique provided higher efficiency and made use of nonedible resource.
In order to produce an alternative energy of biodiesel using field pennycress (Thlaspi arvense L.) seed oil, a novel Fe3O4/graphene oxide-phenylalanine bisulfate ionic liquid core-shell magnetic catalyst was developed. Ferroferric oxide nanoparticles and graphene oxide (GO) were employed to prepare a magnetic carrier of Fe3O4@GO nanocomposite which applied to immobilize phenylalanine bisulfate ionic liquid (PBIL). The immobilized PBIL was characterized by the means of characterization. The heterogeneous Fe3O4@GO@PBIL catalyst was demonstrated to have a high catalytic efficiency that reached 92.38 % of biodiesel yield under the optimal conditions (methanol/oil molar ratio of 10:1, reaction temperature of 60 °C, reaction time of 4 h, catalyst dosage of 25 wt%, stirring speed of 1500 r/min). After transesterification processes, the catalyst could be easily removed from the reaction mixture with an external magnet and showed little deactivation after multiple recycles. The main properties of biodiesel product were reconcilable with the range of test standards of ASTM D6751 and EN 14214. In summary, the prepared Fe3O4@GO@PBIL catalyst behaved a superiorly catalytic performance, which made the produced biodiesel possess the possibility of application to biofuel as an ideal substitution.
Pennycress (
L.) is being domesticated as an oilseed cash cover crop to be grown in the off-season throughout temperate regions of the world. With its diploid genome and ease of directed mutagenesis ...using molecular approaches, pennycress seed oil composition can be rapidly tailored for a plethora of food, feed, oleochemical and fuel uses. Here, we utilized Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology to produce knockout mutations in the
(
) and
(
) genes to increase oleic acid content. High oleic acid (18:1) oil is valued for its oxidative stability that is superior to the polyunsaturated fatty acids (PUFAs) linoleic (18:2) and linolenic (18:3), and better cold flow properties than the very long chain fatty acid (VLCFA) erucic (22:1). When combined with a
(
) knockout mutation,
and
double mutants produced ∼90% and ∼60% oleic acid in seed oil, respectively, with PUFAs in
as well as
single mutants reduced to less than 5%. MALDI-MS spatial imaging analyses of phosphatidylcholine (PC) and triacylglycerol (TAG) molecular species in wild-type pennycress embryo sections from mature seeds revealed that erucic acid is highly enriched in cotyledons which serve as storage organs, suggestive of a role in providing energy for the germinating seedling. In contrast, PUFA-containing TAGs are enriched in the embryonic axis, which may be utilized for cellular membrane expansion during seed germination and seedling emergence. Under standard growth chamber conditions,
plants grew like wild type whereas
single and
double mutant plants exhibited delayed growth and overall reduced heights and seed yields, suggesting that reducing PUFAs below a threshold in pennycress had negative physiological effects. Taken together, our results suggest that combinatorial knockout of
and
may be a viable route to commercially increase oleic acid content in pennycress seed oil whereas mutations in
will likely require at least partial function to avoid fitness trade-offs.
Efforts are underway to domesticate the winter annual oilseed cover crop, pennycress (Thlaspi arvense L.), to aid in overcoming global food security and ecosystem challenges. Several key alleles for ...domestication have already been introgressed; however, the weedy trait of seed dormancy remains. To make pennycress attractive for growers, any traits that could contribute to weed pressure on their fields must be removed. In this study, we have performed multi-faceted experiments to understand and dissect challenges associated with seed dormancy and germination in wild pennycress. Lines were derived by chemical mutagenesis to provide a possible genetic solution. Laboratory and field-based experiments were performed to understand the role of light, seed priming, and gibberellic acid in the wild-type pennycress. These experiments were followed by incorporation of a genotype with a mutation in the TRANSPARENT TESTA 2 gene that could potentially overcome problem of seed dormancy in wild-type pennycress. Subsequent laboratory and field-based comparison of the wild-type and tt2 mutant over three seasons suggested that the tt2 line can germinate at a faster rate than wild-type and is comparable to the germination of wild-type treated with gibberellic acid. These experiments validated the hypothesis of using transparent testa mutants to solve seed dormancy and germination challenges. Further characterization of the transparent testa mutant indicated that this genotype can be susceptible to abiotic stress conditions such as NaCl, polyethylene glycol, coarse growth medium (sand), and abscisic acid. Future work is required to understand the issues related to the abiotic stress response in these lines.
•The germination rate of the tt2 mutant was ≥ 89% versus ≤ 63% for wild-type.•Soil texture and precipitation greatly influenced germination of the tt2 mutant.•Salt, water, and ABA stress reduced tt2 germination compared to wild-type.
Commercial production of jet fuel from biobased feedstocks is still encumbering, mainly due to their high production cost and competition with food resources. Pennycress oilseed is a potential jet ...fuel feedstock which can be supplied at a lower price compared to similar oilseeds, such as soybean and canola. However, the techno-economics of pennycress-based jet fuel production needs to be evaluated. The objective of this study was to assess the technical feasibility and costs of hydroprocessed renewable jet fuel (HRJ) production from pennycress. The production capacity was considered to be 18.9 million L/yr (5 million gal/yr). The analysis considered pennycress grain handling and conditioning, oil extraction and conversion to HRJ and byproducts, i.e., LPG, naphtha and green diesel, through hydroprocessing technology, as well as pennycress meal processing as boiler fuel and wastewater treatment. Total investment for establishing the HRJ biorefinery at the selected capacity was estimated to be 90.8 million USD. Minimum selling price (MSP) of HRJ was estimated to be 1.2 USD/L, which was comparable to the MSP of HRJ from similar oilseeds, including soybean and canola. It could also be further reduced by supplying pennycress grain at a lower price, increasing the oil content and increasing the production capacity of the biorefinery. The outcomes of this research would help establish the performance targets needed to reach the economic viability of HRJ production from pennycress at the commercial scale.
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•Operating cost of jet fuel is dominated by feedstock supply, facility and labor costs.•Minimum selling price of jet fuel from pennycress is comparable to that from similar oilseeds.•Pennycress grain price and biorefinery size mainly affect minimum selling price of jet fuel.•Upgrading pennycress meal to high-value products would improve the economics of jet fuel.
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