Oilseeds may be processed to yield a number of potentially valuable compounds and fractions including oil, protein and small molecules. However, energy costs associated with industrial processing of oilseeds can be significant. For example, processes that use water to dissolve and separate materials are burdened with the costs associated with concentrating value-added products from dilute solutions. The ethanol industry produces large amounts of an aqueous solution called thin stillage that has little value and is used in animal feed. Thin stillage contains some of the necessary salts used in protein extraction but has a low pH. Protein extraction and protein isolate production is commonly conducted at higher pH. Waste alkali from biodiesel production has a high pH and can be used to adjust the pH of thin stillage to improve its ability to extract protein from oilseed meal. By combining the properties of the waste products of both the ethanol and the biodiesel industries, a complementary process is possible that may have greater economic potential than current practices in industry.<p> In this study, processes for protein extraction from mustard (<i>Brassica juncea</i> (L.) Czern.) meal using thin stillage from ethanol production and glycerol from biodiesel production were studied. The osmotic potential of thin stillage used in this research was lower than that of water, whereas both the density and the viscosity were higher. The pH was typically 3.7-3.8, and the total Kjeldahl nitrogen was approximately 0.080.10 %, w/w. Organic compounds identified in thin stillage were isopropanol, ethanol, lactic acid, 1,3-propanediol, acetic acid, succinic acid, glycerophosphorylcholine, betaine, glycerol and phenethyl alcohol. In addition, yeasts, bacteria and fungi were also found. Moreover, the salt types and their concentrations in thin stillage were predictable. The salt types present in thin stillage were CaCl2, NaCl, K2SO4, NaNO3, Mg(OH)2, Na2SO4 and KOH. A model thin stillage synthesized for the purposes of this research had components and chemical and physical properties comparable to those of thin stillage from ethanol production. Protein was extracted from ground, defatted meal using thin stillage at different pHs and salt concentrations. The results showed that pH and salt content affected protein extraction efficiency. However, no differences were found in the efficiency of extraction, SDS-PAGE profile, digestibility, lysine availability or amino acid composition of protein extracted with thin stillage, model thin stillage or sodium chloride solution. Moreover, extracted protein did not display significant hydrolysis. The results from peptide sequencing showed that napin and cruciferin were the most prevalent proteins in the extracted fractions. When increasing the scale of the extraction, the efficiency of protein extraction and the percentage of protein in the extracted protein were decreased. Protein recovery achieved with the complementary protocol was higher than that reported for a published protocol. Allyl isothiocyanate was found in protein extracts.

<p>Ethiopian mustard (<i>Brassica carinata</i> A. Braun) or gomenzer is an oilseed crop that is well adapted to the highlands of Ethiopia. Evaluation of the local germplasm has resulted in the registration of high yielding cultivars, such as Dodolla and S-67. The oil of gomenzer contains about 40% erucic acid and the meal is high in glucosinolates. The objective of this research was to study the inheritance of erucic acid content in gomenzer and to introgress genes for the non2-propenyl glucosinolate trait from <i>B. napus</i> and <i>B. juncea</i>. The erucic acid content of F₁ seed from reciprocal crosses between the high erucic acid cultivars Dodolla and S-67 and zero erucic acid line C90-14 was intermediate between the parents indicating that erucic acid content in B. carinata was controlled by two nondominant genes with two alleles acting in an additive manner. Backcross F₁ seed derived from the backcross to the low erucic acid parent fell into three erucic acid classes with $<$0.5%, 6 to 16% and $>$16% erucic acid at the ratio of 1:2:1 indicating that erucic acid was under the control of two alleles each of at two loci. F₂ seed segregation data supported this observation. Each allele contributed approximately 10% erucic acid. The high glucosinolate B. carinata line C90-14, low glucosinolate <i>B. napus</i> cultivar Westar and <i>B. juncea</i> line J90-4253 were chosen as parents for the development of non2-propenyl glucosinolate <i>B. carinata</i>. The objective was to transfer genes for non2-propenyl glucosinolate content from <i>B. napus</i> and <i>B. juncea</i> into <i>B. carinata.</i> Interspecific crosses were made between <i>B. carinata</i> and <i>B. napus</i>, <i>B. carinata</i> and <i>B. juncea</i> and the interspecific F₁ generations were backcrossed to <i>B. carinata</i>. Backcross F₁ plants from the two interspecific crosses were intercrossed in an attempt to combine the two sources for non2-propenyl glucosinolate content in one genotype. Seed of backcross F₁ plants of the cropss ((<i>B. carinata</i> x <i>B. napus</i>) x <i>B. carinata</i>) contained a high concentration of 2-propenyl glucosinolate similar to those of <i>B. carinata</i>. Introgression of C genome chromosomes of <i>B. napus</i> into <i>B. carinata</i> was not effective in redirecting glucosinolate synthesis away from 2-propenyl and into 3-butenyl glucosinolate. This indicated that C genome chromosomes do not contain genetic factors for C3 $\to$ C4 glucosinolate precursor chain elongation, and that 2-propenyl glucosinolate synthesis is primarily controlled by genes on B genome chromosomes. Seed of ackcross F₂ plants of the cross ((<i>B. carinata</i> x <i>B. juncea</i>) x <i>B. carinata</i>) contained much reduced levels of 2-propenyl glucosinolate indicating that genetic factors for C3 $\to$ C4 glucosinolate precursor chain elongation were introgressed from the B genome of <i>B. juncea</i> into the B genome of <i>B. carinata</i>. However, a complete diversion of glucosinolate synthesis from 2-propenyl to 3-butenyl was not achieved. Further selections in segregating F₄ and F₅ generations of <i>B. juncea</i> derived <i>B. carinata</i> populations could yield the desired zero 2-propenyl glucosinolate B. carinata. The double interspecific cross was unsuccessful.

Oilseeds may be processed to yield a number of potentially valuable compounds and fractions including oil, protein and small molecules. However, energy costs associated with industrial processing of oilseeds can be significant. For example, processes that use water to dissolve and separate materials are burdened with the costs associated with concentrating value-added products from dilute solutions. The ethanol industry produces large amounts of an aqueous solution called thin stillage that has little value and is used in animal feed. Thin stillage contains some of the necessary salts used in protein extraction but has a low pH. Protein extraction and protein isolate production is commonly conducted at higher pH. Waste alkali from biodiesel production has a high pH and can be used to adjust the pH of thin stillage to improve its ability to extract protein from oilseed meal. By combining the properties of the waste products of both the ethanol and the biodiesel industries, a complementary process is possible that may have greater economic potential than current practices in industry.<p> In this study, processes for protein extraction from mustard (<i>Brassica juncea</i> (L.) Czern.) meal using thin stillage from ethanol production and glycerol from biodiesel production were studied. The osmotic potential of thin stillage used in this research was lower than that of water, whereas both the density and the viscosity were higher. The pH was typically 3.7-3.8, and the total Kjeldahl nitrogen was approximately 0.080.10 %, w/w. Organic compounds identified in thin stillage were isopropanol, ethanol, lactic acid, 1,3-propanediol, acetic acid, succinic acid, glycerophosphorylcholine, betaine, glycerol and phenethyl alcohol. In addition, yeasts, bacteria and fungi were also found. Moreover, the salt types and their concentrations in thin stillage were predictable. The salt types present in thin stillage were CaCl2, NaCl, K2SO4, NaNO3, Mg(OH)2, Na2SO4 and KOH. A model thin stillage synthesized for the purposes of this research had components and chemical and physical properties comparable to those of thin stillage from ethanol production. Protein was extracted from ground, defatted meal using thin stillage at different pHs and salt concentrations. The results showed that pH and salt content affected protein extraction efficiency. However, no differences were found in the efficiency of extraction, SDS-PAGE profile, digestibility, lysine availability or amino acid composition of protein extracted with thin stillage, model thin stillage or sodium chloride solution. Moreover, extracted protein did not display significant hydrolysis. The results from peptide sequencing showed that napin and cruciferin were the most prevalent proteins in the extracted fractions. When increasing the scale of the extraction, the efficiency of protein extraction and the percentage of protein in the extracted protein were decreased. Protein recovery achieved with the complementary protocol was higher than that reported for a published protocol. Allyl isothiocyanate was found in protein extracts.

Thesis (Ph. D.)--University of California, Riverside, 2009. / Includes abstract. Title from first page of PDF file (viewed Febrary 1, 2010). Includes bibliographical references (p. 140-143). Issued in print and online. Available via ProQuest Digital Dissertations.

The thesis addresses the potential application of ruthenium(II)-cobalt(III) heterodinuclear complexes as a new selective cancer treatment. The selectivity is to be achieved through the use of visible light to trigger activation of the drug. The majority of work conducted relates to the design and synthesis of the bridging ligand for the final ruthenium(II)-cobalt(III) heterodinuclear complex. In Chapter 2, a potential bridging ligand based on a functionalised terpyridine is described. The intention was to bind the ruthenium(II) metal centre to the terpyridine end of the bridging ligand and have a secondary binding domain available for coordination of the cobalt(III) metal centre. However, a reductive step in the synthetic pathway failed to produce the desired product and this potential bridging ligand had to be abandoned. In Chapter 3, two series of bridging ligands are described. The first of these series is based on Jurgen Sauer’s ‘LEGO’ system. In addition to describing the free synthesis of these ligands, their synthesis on a ruthenium(II) metal centre is described. The second series is based on disubstituted-1,2,4,5-tetrazines. These compounds are only able to be directly synthesised as the non-coordinated ligand. Coordination of these ligands to a single ruthenium(II) metal centre is then described. Ruthenium(II) complexes of both ligand series are then exposed to several transition metals and their ability to coordinate a second metal centre investigated. The formation of ruthenium(II)-cobalt(III) heterodinuclear complexes, using the ligand series detailed in Chapter 3, is described in Chapter 4. These complexes are formed by reacting the ruthenium(II) complex of the bridging ligand with either [Co(en)₂(OTf)₂](OTf) or [Co(tren)(OTf)₂](OTf). These heterodinuclear complexes exhibit photo-activated ligand release, which makes them candidates for development as a potential cancer treatment. The non-bridging ligands coordinated to the cobalt(III) metal centre in Chapter 4 were not cytotoxic. In order to make the system biologically active these ligands need to be changed. Chapter 5 describes how nitrogen mustards (a class of cytotoxic DNA alkylators) could be introduced as the non-bridging ligands. This involves the synthetic strategy of forming the cobalt(III) complex of the alcohol precursor of a nitrogen mustard. This precursor complex is then converted into the nitrogen mustard complex and coordinated to the ruthenium(II) bound bridging ligand. The synthetic strategies outlined in this thesis can be applied to a wide range of potential bridging ligands and could potentially lead to a large number of ruthenium(II)-cobalt(III) heterodinuclear complexes being synthesised. One journal article based on this research has been accepted for publication, in the Australian Journal of Chemistry. Three more articles are in preparation.

Ready-to-eat meat products such as dry-fermented sausages have been associated with foodborne outbreaks despite the multiple hurdles used in the manufacturing process to prevent growth of pathogens. As a result, new strategies such as natural products with antimicrobial activity are being used to control pathogens of importance like Escherichia coli O157:H7. This study investigated how different concentrations and sources of mustard can influence its antimicrobial activity against E. coli O157:H7 in dry-fermented sausage, as well as the contribution of residual myrosinase enzyme in mustard to this process. The genetic basis for the degradation of mustard glucosinolate by E. coli O157:H7, which is associated with the antimicrobial action of mustard, was also characterized. The ability of E. coli O157:H7 to withstand inhibitory allyl isothiocyanate (AITC) concentrations and the role of the two-component BaeSR system as a defense mechanism against AITC was also investigated. Results showed that 4% (w/w) deodorized yellow mustard powder was effective to control E. coli O157:H7 in dry-fermented sausage at 28 d. The presence of endogenous plant myrosinase in the mustard powder or meal enhanced E. coli O157:H7 reduction rates. Fully-deodorized, deoiled, yellow mustard meal as low as 2% (w/w) containing either 0.1% or 0.2% of residual plant myrosinase achieved the same results as 4% (w/w) mustard powder also containing similar residual myrosinase. Regardless of the type of mustard, the antimicrobial activity of yellow mustard derivatives were more pronounced than those of Oriental mustard. The initial genetic assessment through in silico analysis found similarity between plant myrosinase and enzymes encoded by genes (bglA, ascB, and chbF) from β-glucosidase families in E. coli O157:H7 strains. After disruption of these genes using lambda-red replacement, single (∆bglA, ∆ascB, ∆chbF) and double (∆bglAascB, ∆chbFascB, ∆chbFbglA) mutant strains were created and assessed for glucosinolate degradation. The comparison of the gene expression profiles and changes in the extent of sinigrin degradation by different mutants suggested that ascB have a prominent role in the degradation of this β-glucoside by E. coli O157:H7. E. coli O157:H7 did not develop resistance to AITC, the essential oil formed from sinigrin degradation that is responsible for the antimicrobial activity of Oriental mustard.

Canola meal (Brassica napus L.) and oriental mustard bran (Brassica juncea) were subjected to alkali and acid pretreatment to expose pentosan, for enhancing further enzymatic hydrolysis by endo-1,4-β-xylanase from Trichoderma longibrachiatum for the production of oligosaccharides. Pretreatment especially with alkali, effectively increased the relative content of pentosan to about ~ 41% and ~ 72%. Alkali pretreated canola meal and mustard bran resulted in a pentose content of 2.28 ± 0.15 g and 3.20 ± 0.11 g per 100 g substrates at 18 h and 24 h of reaction respectively, which corresponded to ~ 26% and ~ 28% conversion of original pentosan in substrates. UPLC-MS data showed xyloglucuronic acid (XGlcA) as the major oligosaccharide in the hydrolyzates. Reversed-phase HPLC-DAD indicated the principal phenolic compound in the hydrolyzates was sinapine. DPPH radical scavenging assay showed that endoxylananse hydrolyzates of acid pretreated substrates had strong antioxidant activities in comparison to alkali pretreated samples.

Two types of protein isolates were prepared from de-hulled yellow mustard flour by aqueous extraction, membrane processing and acid precipitation of proteins at the isoelectric point (IEP 5.5). Their electrophoretic, main functional properties and protein composition were determined. The precipitated and acid soluble protein isolates had 83.0 and 96.0% protein content on a moisture and oil free basis, respectively. The acid soluble protein isolate had comparable functional properties to those of commercially available soybean and other protein isolates. The precipitated protein isolate exhibited less desirable functionality than the soluble isolate, due to its high lipid content (~25%); however, it was still comparable to soybean isolates. Storage temperature had limited effect on lipid oxidation, and hence the stability of the precipitated protein isolate at 25-45ºC. Taste and texture of wieners and bologna prepared with 1-2% of this isolate as binder were comparable to those prepared with soy protein isolates.

Two types of protein isolates were prepared from de-hulled yellow mustard flour by aqueous extraction, membrane processing and acid precipitation of proteins at the isoelectric point (IEP 5.5). Their electrophoretic, main functional properties and protein composition were determined. The precipitated and acid soluble protein isolates had 83.0 and 96.0% protein content on a moisture and oil free basis, respectively. The acid soluble protein isolate had comparable functional properties to those of commercially available soybean and other protein isolates. The precipitated protein isolate exhibited less desirable functionality than the soluble isolate, due to its high lipid content (~25%); however, it was still comparable to soybean isolates. Storage temperature had limited effect on lipid oxidation, and hence the stability of the precipitated protein isolate at 25-45ºC. Taste and texture of wieners and bologna prepared with 1-2% of this isolate as binder were comparable to those prepared with soy protein isolates.

Thesis (M.S.)--University of Toledo, 2009. / Typescript. "Submitted as partial fulfillment of the requirements for The Master of Science Degree in Biology (Ecology-track)." "A thesis entitled"--at head of title. Bibliography: leaves 28-32.