Thesis (MTech (Food Technology))--Cape Peninsula University of Technology, 2018. / The aim of this study was to produce Bambara groundnut (BGN) starch-catechin complex using chemical initiators (ascorbic acid and hydrogen peroxide) and cyclodextrin (alpha and beta) with the view to obtain a functional ingredient for the food industry. BGN starch was successfully extracted from BGN flour through dry milling method, yielding 32% of BGN starch. Native BGN starch was chemically modified using ascorbic acid (1% w/w) and hydrogen peroxide (165% w/w) as redox, biocompatible initiator for grafting catechin to the BGN starch. In addition, cyclodextrin (alpha and beta) were also used as initiators for modifying BGN starch through complexation methods. Complexation methods used included the microwave, co-evaporation and kneading. The characterization of native and modified BGN starches was carried out by performing scanning electron microscopy, powder X-ray diffraction, Fourier Transform Infrared (FTIR) and fluorescence spectroscopy analysis. Functional, thermal and rheological properties of native and modified BGN starches were evaluated. The pasting properties of BGN starches were determined using the Rapid Visco Analyser (RVA). According to the SEM profile, native BGN starch had round, oval and elliptical shapes typical for legume starches. Native BGN starch displayed a typical type-C crystallinity which is common among legumes with strong peaks at 2θ of 15o, 17o and 23o. BGN starches modified through complexation methods had sharper peaks indicating an increase in starch crystallinity; however, following chemical modification there was loss in starch crystallinity which was evidenced by the amorphous region in the chemically modified BGN starches. Structure of native and modified BGN starches was confirmed by FTIR. The FTIR spectra of native BGN starch showed variable peaks at 3285.34 cm-1, 2931.69 cm-1, 1634.36 cm-1, 1336.77 cm-1 which are attributed to OH stretching, C-H stretching, water bending vibrations and C-O stretching, respectively. Furthermore, the FTIR results confirmed that native BGN starch is made up of glucose molecules just like all other starches. All modified BGN starches displayed a new absorption peak at 1020 cm-1 wavelength, thus indicating that starch modification was successful. On the other hand, all BGN starch-catechin complexes displayed a new absorption peak in the range of 1520 -1560 cm-1, attributed to the C-C stretching within the aromatic ring of the catechin. The successful grafting of catechin to BGN starch was also confirmed by the fluorescence spectroscopy results, where all the BGN starch-catechin complexes had an emission peak at 320 nm while native BGN starch had an emission peak at 270 nm. Antioxidant capacity of BGN starch was determined through DPPH and ORAC antioxidant assays. Within the DPPH assay, the antioxidant activity ranged from 2.26 to 38.31 μmol TE/g. The antioxidant activity of modified BGN starch-catechin complexes was significantly (p ≤ 0.05) higher than the ones modified without catechin. On the other hand, within the ORAC assay, the antioxidant activity ranged from 0.07 to 126.71 μmol TE/g. As opposed to the results obtained in DPPH assay, the antioxidant activity of chemically modified BGN starch-catechin complexes was significantly (p ≤ 0.05) higher than that of complexed BGN starch-catechin complexes. Chemical modification significantly increased the swelling capacity of native BGN starch while complexation methods significantly reduced it.

Thesis submitted in fulfilment of the requirements for the degree Master of Technology: Food Technology in the Faculty of Applied Sciences at the Cape Peninsula University of Technology / The aim of this study was to evaluate bambara groundnut [BGN] non-starch polysaccharides [NSP] subject to the incorporation into model and food systems with a view to establish their functional and physicochemical properties. BGN insoluble dietary fibre [BGNIF] and soluble dietary fibre [BGNSF] were successfully extracted from four varieties (black-eye: BLE, red: RED, brown: BRN and brown-eye: BRE). Physicochemical properties evaluated revealed the high bulk density of all BGNIF and BGNSF varieties, which could contribute to cost-effective packaging. The microstructures of BGNIFs were irregular in shape with different sizes. The colour parameters (lightness, redness, yellowness, chroma and hue angle) differed significantly [p ≤ 0.05] across all BGNIF and BGNSF varieties; and indicated a yellowish-red colour for BGNIFs and a light yellow colour for BGNSFs. Negligible amounts of condensed tannins [CT] were found in BGNIFs (0.014 – 0.160 mg.g-1). Higher amounts polyphenols [PP] were present in BGNSFs (45.42 – 55.90 mg.g-1 gallic acid equivalents [GAE]) compared to the amount PP in BGNIFs (6.14 – 15.56 mg.g-1 GAE). Major sugars identified were arabinose/galactose, xylose and mannose in BGNIFs, and xylose and mannose in BGNSFs. The functional properties evaluated revealed high swelling capacity of BGNIFs (6.37 – 7.72 ml.g-1) and no significant [p > 0.05] difference in water retention capacity. Fat absorption capacity ranged from 1.38 – 1.52 g oil.g-1 dry weight for BGNIFs and 4.04 – 4.55 g oil.g-1 dry weight for BGNSFs. Variability in BGNIF (91.2%) and BGNSF (79.4%) physicochemical and functional properties could both be explained by two principal components (BGNIF component 1: PP, redness, yield; and component 2: xylose, yellowness and chroma; BGNSF component 1: yellowness, chroma, mannose content; and component 2: redness, fat absorption and fructose content). Following an IV optimal mixture design, an optimum white bread formulation was obtained using 59.5% water, 4.3% yeast and 8.5% BGNIF. Bread enriched with the four BGNIF varieties (BLE, RED, BRN and BRE) were tested for several physicochemical properties. Significant [p ≤ 0.05] differences existed between the control and BGNIF enriched loaves for crumb grain characteristics (including pore area distribution, feret angle, circularity, roundness and aspect ratio). Specific loaf volume of BGNIF enriched loaves ranged from 3.33 – 3.85 ml.g-1 and were significantly [p ≤ 0.05] lower compared to the control bread (4.16 ml.g-1). Favourable texture characteristics obtained with the BGNIF enriched breads were lower hardness, chewiness and gumminess compared to the control loaf. Crust and crumb colour parameters (lightness, redness, yellowness, chroma and hue angle) were significantly [p ≤ 0.05] different across all loaves. BRE BGNIF bread (3.43 ± 0.20) had the significantly [p ≤ 0.05] lowest crumb colour difference compared to the control bread; whilst BRN (1.72 ± 0.42) and BRE (2.44 ± 0.78) loaves had the lowest significant [p ≤ 0.05] crust colour difference compared to the control. Favourable chemical properties were the high total dietary fibre [TDF] (7.14 – 8.33%) content of all BGNIF enriched loaves compared to the control loaf (4.96%). Significant [p ≤ 0.05] differences were also observed for some loaves for moisture content, condensed tannins and polyphenol content. Variability in bread physicochemical properties was differentiated by three components (component 1: bread textural properties; component 2: specific loaf volume and bread lightness; component 3: crumb colour parameters) which accounted for a cumulative variation of 92.8%. All bread loaves were also sensorially acceptable as rated moderately like to like very much (>3 rating on a 5-point hedonic scale) by consumers for all parameters (appearance, crust and crumb colour, aroma, taste, texture and overall acceptability) evaluated. Furthermore, brown BGNSF was tested for stabilising effects in an orange beverage emulsion. BGNSF and orange oil were varied at two levels each based on a 22 augmented factorial design and the effects determined on the equilibrium backscattering [BS] flux as emulsion stability indicator. The BS profiles which resulted from the Turbiscan stability analysis revealed flocculation at low rates as the major destabilisation mechanism. The optimal formulation producing a stable emulsion was identified as low oil (6%) and high BGNSF (30%) concentrations. The objective of this study was therefore achieved and showed that positive physicochemical and functional properties are associated with BGNIF and BGNSF from black-eye, red, brown and brown-eye varieties. Furthermore, the incorporation of BGN fibres in white bread and a beverage emulsion was shown to contribute positive technological properties in these systems.

Bambara groundnut (Vigna subterranea [L.] Verdc.) is an under-utilized indigenous African legume crop which has substantial potential to contribute to food security in sub-Saharan Africa. The crop is well adapted to severe agro-ecologies and grows where other legumes may not survive. The seed is highly nutritious with an ideal balance of carbohydrate (55-72%), protein (18-20%) and fats (6-7% oil), which is particularly beneficial in balancing protein deficiencies in cereals. Also, the seed contains essential and non-essential amino acids of about 33% and 66%, respectively. These attributes make Bambara groundnut an ideal crop to alleviate food insecurity, and to reduce protein malnutrition in rural communities of Africa. However, small-scale farmers grow low-yielding landraces in most production regions in sub-Saharan Africa. Bambara groundnut landraces exist as heterogeneous mixtures of seeds of a few to several seed morpho-types that embrace wide genetic potential for breeding. The objectives of this study were: 1) to determine the production status and constraints associated with Bambara groundnut production in Kano State of Nigeria through the use of a participatory rural appraisal (PRA); 2) to determine the genetic diversity of Bambara groundnut landraces through seed morphology; 3) to assess the inter- and intra-genetic diversity of the Bambara groundnut landraces; 4) to determine the yield and yield component responses among selected Bambara groundnut genotypes, 5) to determine the genomic diversity in Bambara groundnut landraces, using simple sequence repeat (SSR) markers; and 6) to develop a crossing protocol. Using a structured questionnaire, 150 Bambara groundnut farmers were interviewed. The respondents interviewed were male and aged between 36 to 50 years, while Qur’anic education was the most popular among them. Most of the farmers practiced a combination of sole and mixed cropping, and allocated between 0.38 to 1.68 hectares of land to Bambara groundnut growing. They selected Bambara groundnut landraces, especially looking for large seeds that were pure and oval in shape, with a cream seed coat colour and which were early maturing. A total of 27 diverse landraces bearing different names were identified in the hands of the farmers. Most popular among them were Gurjiya, Kurasa, Hawayen-Zaki, Fara Mai-Bargo and Silva. Production was largely for home consumption and for sale on local markets. Common production constraints of the crop were identified as a lack of improved varieties (70.7%), frequent droughts (9.3%), low yield (4%) and limited access to large markets (3.3%). Diverse collections of Bambara groundnut landraces from seven geographic origins were characterized using seed morphology, including seed coat, seed eye colour and pattern, and hilum colour and pattern. Out of 58 original seedlots, a total of 353 different seed morpho-types were further identified. The selected morpho-types- can be used for large-scale production or true-to-type lines could be used in genetic improvement of the crop. Genetic variability within- and between-landraces was investigated among 262 Bambara groundnut landraces, forty nine were studied for agronomic traits, and 213 were investigated for pod and seed variability. Most (47.9%) of the landraces developed pods with a point on one pole, and a round end on the other. Most had a creamy (37.1%) and yellow (76.1%) pod colour, and the pods were usually rough textured, and contained an oval seed. A further 158 landraces were evaluated for leaf morphology where 49.4% had round leaves, while 21.5% had elliptic leaves, with 55.7% of the landraces being heterogeneous, possessing more than one form of leaf shapes. These discrete characters can be utilized for genetic studies and improvement of Bambara groundnut. Single plant selections of 49 Bambara groundnut genotypes were evaluated for yield and yield components using 26 yield and yield related traits. Highly significant variations (P<0.001) were detected among the genotypes for canopy spread, petiole length, weight of biomass, seed weight and seed height. Principal component analysis (PCA) identified nine useful components, where two components, PC1 and PC2, contributed strongly to the total variation, at 19% and 14%, respectively. The PCA revealed that leaf colour at emergence, petiole colour, leaf joint pigmentation and calyx colour were highly correlated with PC1, while seed length, seed width and seed height had strong association with PC2. Both the principal component and cluster analyses showed that most genotypes associated with one another with respect to agronomic and seed yield traits, irrespective of geographical location. The genotypes 211-57, MO9-4 and TV-27 displayed high seed yield performances, while TV-93 and 45-2 had higher biomass production. These genotypes can be used as breeding lines to enhance productivity of Bambara groundnut. Fifty Bambara groundnut genotypes, representing seven geographical regions across Africa, were genotyped using five pre-selected polymorphic simple sequence repeat (SSR) markers developed specifically for Bambara groundnut. The results detected a total of 53 alleles among the 50 Bambara groundnut genotypes, while the neighbor-joining analysis generated seven major genetic groups, which were clustered regardless of their geographic origin. Close relationship were found between 211-68 on one hand and 211-83-2, N211K and M09-3 with 211-68 on the other. Genotypes M02-3, 211-55-1 and 211-57 displayed close similarities. These associations suggested the likelihood that the two pair groups had common origins or may possess similar genes. A preliminary protocol was developed for crossing Bambara groundnut using eight selected parents, using the diallel mating system. Emasculation and crossing of Bambara groundnut was effective when conducted on the same day, with the two procedures being carried out sequentially between 4:30 am and 9:00 am. This protocol generated a number of F1 seeds, with the most success being from crosses between 211-40-1 x N211-2, N212-8 x 211-40-1 and M09-3 x 211-82-1. These F1 seeds can be advanced to confirm whether they are true F1 or selfs. The most important production constraint of Bambara groundnut production is the lack of improved varieties, suggesting that further breeding is needed to enhance productivity. Bambara groundnut landraces need to be sorted using discrete morphological features before breeding for genetic enhancement. The SSR markers used in the study demonstrated their ability to distinguish the existing diversity among the Bambara groundnut genotypes, which could be useful for both germplasm conservation and for breeding. Genotypes that displayed outstanding performance in seed yield and biomass can be used as breeding lines for the genetic improvement of Bambara groundnut. Overall, the study generated valuable and novel Bambara groundnut genetic material, useful in the development of improved cultivars for large-scale production in sub-Saharan Africa. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2014.

Thesis (MTech (Agriculture))--Cape Peninsula University of Technology, 2014. / Plants require nitrogen (N) to support desired production levels. Nitrogen fertilization strategy is a major consideration in field management with regard to achieving both economic and environmental objectives. For instance, in potato, insufficient N supply reduces tuber size and overall yield while excessive N supply can reduce tuber quality and increase environmental risk through nitrate (NO3-) leaching and nitrous oxide emission. Selection of an adequate N fertilizer application rate for crops is difficult, due to marked variations in soil N supply and crop N demand in both the field and over time. This research was conducted to characterise the ammonium transporter gene (AMT1) of Bambara groundnut and potato using molecular biology and bioinformatics methods. Nucleotide database sequences were used to design AMT1-specific primers which were used to amplify and sequence the core-region of the gene from Bambara groundnut and potato. Bioinformatics techniques were used to predict the structure and infer properties of the proteins. Nucleotide sequence alignment and phylogenetic analysis indicate that BgAMT1 and PoAMT1 are indeed from the AMT1 family, due to the clade and high similarity they respectively shared with other plant AMT1 genes. Amino acid sequence alignment showed that BgAMT1 is 92%, 89% and 87% similar to PvAMT1.1, GlycineAMT1 and LjAMT1.1 respectively, while PoAMT1 is 92%, 83% and 76% similar to LeAMT1.1, LjAMT1.1 and LeAMT1.2 respectively. BgAMT1 and PoAMT1 fragments were shown to correspond to the 5th - 10th transmembrane spanning-domains. Mutation of Bg W1A-L and S28A (for BgAMT1) and Po S70A (for PoAMT1) is predicted to enhance ammonium (NH4+) transport activity. Residues Bg D23 (for BgAMT1) and Po D16 (for PoAMT1) must be preserved otherwise NH4+ transport activity is inhibited. In all, BgAMT1 and PoAMT1 play a role in N uptake from the root while BgAMT1 may contribute more in different steps of rhizobia interaction. In an investigation of the correlation between AMT1 gene expression levels and leaf chlorophyll content index (CCI) with plant N status, potato plants were grown in a hydroponic greenhouse with 0.75 or 7.5 mM NO3- and 0.75 or 7.5 mM NH4+ as forms of N supply in a completely randomized design. Leaf CCI as measured by chlorophyll content meter, showed that an increase in N supply results in increased leaf CCI in response to both forms of N. Total RNA was isolated from leaf sampled at 28 days after treatment and expression level of the AMT1 gene was determined by reverse transcription-qPCR using a second set of primers designed for qPCR. The results showed that expression levels of AMT1 increased from 8.731 ± 2.606 when NO3- supply was high to 24.655 ± 2.93 when NO3- supply was low. However, there was no significant response in AMT1 expression levels to changes in NH4+. This result suggested that AMT1 transports NO3- less efficiently than NH4+, and thus more transport channels are required in the cell membrane when NO3- levels are low. Such variation in AMT1 expression levels are not necessary for NH4+ transport since the transport mechanism for NH4+ is efficient even at low NH4+ levels.

Submitted in fulfillment of the academic requirement for the degree Doctor of Philosophy (Ph.D.) in Food Science and Technology, Durban University of Technology, Durban, South Africa, 2017. / Bambara groundnut (Vigna subterranea) is an indigenous underutilised leguminous crop to Africa. It is a good source of protein and carbohydrate including starch. Bambara groundnut is a traditional crop grown mainly for subsistence in Southern Africa. Bambara groundnut has the advantage of being drought tolerant and can thrive in hot temperatures and poor soil conditions. Therefore, it has great potential as an alternative crop to soya bean and peanuts for cultivation and utilisation. Bambara groundnut starch can potentially be used for various industrial applications. However, native starches are not suitable for most industrial applications, hence the need for modification. Bambara groundnut starch has been previously modified using physical and chemical modification methods. Natural alternatives such as the use of lipids are being sought to modify starches due to the associated risk with chemically modified starch. In this research, Bambara groundnut starch was modified with lipids to improve functional properties, utilisation and application. Specifically, the physicochemical properties of native Bambara groundnut starch obtained from five Bambara groundnut genotypes and three landraces (maroon, brown and cream) were determined. Bambara groundnut starch was modified with lipids (palmitic acid, stearic acid, oleic acid, linoleic acid and lysophosphatidylcholine) and the physicochemical properties of the modified starch were investigated. Further, the influence of high-pressure homogenization on complexation of Bambara groundnut starch with lipids was assessed in comparison with maize and potato starches. Lastly, an application of modified Bambara groundnut starch in biofilm production was also studied. Bambara groundnut landraces generally showed higher amylose contents (approx. 33%) than the genotypes (approx. 28%). Differences were observed in the crystalline patterns of these starches. Bambara groundnut genotypes exhibited the C-type-crystallinity, while the landraces showed the unusual A-type pattern. In terms of functionality, landrace starches showed better swelling than the genotypes. Subsequent studies on modification used maroon Bambara groundnut starch since the amylose content was higher than other landraces and there was a consistent supply of the grains during the period of the study. Generally, Bambara groundnut starch showed higher complexing ability with all the lipids than maize and potato reference samples. These differences in complexing ability among the starches could be due to the variation in amylose contents (Bambara groundnut starch: 31.5%, maize: 22.5% and potato: 24.6%). Fatty acids complexed better with Bambara groundnut starch than lysophosphatidylcholine, which could be due to the structural differences in comparison with the lysophosphatidylcholine molecule. The number of fatty acid in the glycerol backbone and the additional steric hindrance of the polar phosphatidic acid group in the lysophosphatidylcholine may have reduced its complexing ability. Among the fatty acids, palmitic acid complexed better than stearic and the unsaturated fatty acids, possibly due to its short chain length compared to other fatty acids. Bambara groundnut starch showed reduced peak and setback viscosities in the presence of stearic acid, linoleic acid and lysophosphatidylcholine, suggesting the formation of V-amylose complex. Bambara groundnut starch pasted with lipids displayed reduced gelling ability compared to their unmodified counterparts. XRD studies of freeze-dried paste revealed peaks at 2Ѳ = 7.4, 12.9 and 19.9o confirming the formation V-amylose complexes in Bambara groundnut starch. Modification of Bambara groundnut starch with lipids resulted in reduced digestibility. High-pressure homogenization significantly increased the complexing ability of Bambara groundnut starch with lipids. Homogenized Bambara groundnut starch-lipid complexes generally exhibited higher complex index than their unhomogenized counterparts. The higher complexing ability could be attributed to the effect of high-pressure which may have enhanced greater dispersion of lipids in the starch-water system. X-ray diffraction studies also revealed the formation of higher complexes as shown by high intensities at peaks (2Ѳ= 7.4, 12.9 and 19.9o) corresponding to V-amylose complexes. Bambara groundnut starch-lipid complexes displayed significantly higher melting temperatures (95.74-103.82oC) compared to native uncomplexed starch (77.32oC). Homogenized Bambara groundnut starch complexes were non-gelling while the unhomogenized types produced weak gels, with G′ ˃ G″ in the range of 0.1- 10 Hz. Complexation of Bambara groundnut starch with lipids using high-pressure homogenization may be employed in the production of modified starch with non-gelling properties and higher thermal stability suitable for certain industrial application, such as fat replacers in mayonnaise, frozen foods and desserts for a better mouth feel. The physicochemical and mechanical properties of biofilm prepared from Bambara groundnut starch modified with stearic acid at varying concentrations of 0, 2, 4, 6, 7 or 10% were further studied. By SEM, Bambara groundnut starch films containing stearic acid (˃ 2%) showed a progressively rough surface compared to those with 2% stearic acid and the control. The addition of 2% stearic acid to Bambara groundnut starch film reduced water vapour permeability by approximately 17%. However, mechanical properties of starch films were generally negatively affected by stearic acid. Bambara groundnut starch film may be modified with 2% stearic acid for improved water vapour permeability and thermal stability with minimal effect on tensile strength. / D

Bambara groundnut (Vigna subterranea L. Verdc) is a drought tolerant African legume capable of producing reasonable yields where other crops may fail. However, it remains an underutilised crop, owing to limited research, cultivated using landraces, of which scant information is available describing their agronomy and genetic diversity. The aim of this study was to evaluate the response of bambara landraces from different geographical locations to water stress under controlled and field conditions. Seeds were sourced from subsistence farmers of Tugela Ferry and Deepdale in KwaZulu-Natal (South Africa) and Zimbabwe, and characterised into three seed coat colours: light-brown, brown and red. Seed quality was assessed using the standard germination test. Vigour indices of germination velocity index and mean germination time were determined. Seedling establishment was evaluated using seedling trays using a factorial experiment, with four factors: 1. provenance – (Tugela Ferry and Deepdale), 2. seed colour – (red, light-brown and brown), 3. water regimes – (30%, 60% and 100% field capacity), and 4. soil media – (clay, sand and clay + sand). Seedling leaf samples were used to evaluate proline accumulation as an indicator of stress tolerance. A field trial was used to evaluate productivity of bambara landraces under rainfed and irrigated conditions. A pot trial was conducted under controlled environment conditions with three factors: temperature (33/27°C and 21/15°C), water regimes (30% and 100% of crop water requirement) and bambara landrace selections. Results showed no significant differences in germination capacity between bambara landrace selections. Germination time differed significantly (P<0.001) between bambara landrace selections. The Jozini provenance was shown to perform best, followed by Zimbabwe, Tugela Ferry and Deepdale. Brown landrace selections had higher (P<0.001) germination compared with red and light-brown selections, respectively. Seedling establishment showed that emergence was higher (P<0.001) at 100% FC compared with 60% FC and 30% FC. Emergence was higher (P<0.001) in the Sand+Clay mixture compared with Clay and Sand media. Dark-coloured selections had higher (P<0.001) emergence compared with light-coloured selections. Results from the field trial showed that the red landrace selections emerged better (P<0.001) than the light-brown and brown landrace selections, respectively. Plant growth was lower under irrigated compared with rainfed conditions. Stomatal conductance was higher (P<0.001) under irrigated compared with rainfed conditions, whereas chlorophyll content index was higher (P<0.05) under rainfed compared with irrigated conditions. Results of the pot trial showed that emergence was significantly (P<0.001) affected by temperature. It was higher at 33/27°C compared with 21/15°C (P<0.001). Dark-coloured landraces had higher emergence compared with the light-brown landraces. Stomatal conductance was lower at 30% ET relative to 100% ETc. There were no significant differences between water regimes with respect to biomass, pod number per plant, pod mass per plant, seed number per pod, seed mass per plant and harvest index. It is concluded that seed colour is an important variable in the identity of bambara landraces. Provenance plays a significant role in seed performance and there is a significant interaction between provenance and seed coat colour. This study could be expanded to obtain more data for crop improvement through inclusion of many sites and seasons for better agronomic advice to farmers. / Thesis (M.Sc.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

Thesis (MTech (Food Technology))--Cape Peninsula University of Technology, 2015. / The aim of this study was to characterise the physicochemical, rheological, prebiotic and emulsion stabilising properties of four varieties (black-eye, brown-eye, brown and red) of Bambara groundnut (BGN) extracted using the modified wet milling method. A relatively high yield of BGN dietary fibres was obtained with soluble dietary fibres (SDFs) ranging from 15.4 to 17.1% and insoluble dietary fibres (IDFs) ranging from 12.0 to 15.6%. Black-eye and brown-eye dietary fibres showed superiority in terms of swelling capacities, water holding capacities, oil binding capacities, antioxidant properties as well as thermal stabilities than red and brown dietary fibres. In addition, black-eye and brown-eye dietary fibres were characterised by higher lightness (L*), redness (+a*), yellowness (+b*), chroma (C*) and hue. All four SDFs showed acceptable colour differences with ΔE < 8 ranging from 0.81 to 3.08. The hydrolysable polyphenolic (HPP) content of SDFs ranged from 6.89 to 20.86 mg/g GAE and that of IDFs ranged from 10.96 to 14.43 mg/g GAE. All four SDFs differed significantly (p < 0.05) in their HPP content. BGN IDFs were very low in tannins (< 2.2 mg/g).

Thesis MTech (Food Technology))--Cape Peninsula University of Technology, 2017. / Bambara groundnut (Vigna subterranea) an indigenous legume cultivated in Sub-Saharan Africa has been proclaimed to have medicinal properties from communities and in rural areas. However, there is not enough scientific information to validate these claims. Therefore, this study aimed to identify possible medicinal properties of Bambara groundnut (BGN), by analysing the phytochemical and antimicrobial properties of BGN seed and product extracts from Mpumalanga province within South Africa. The BGN extracts (70% methanol, 70% ethanol, milli-Q water) from seeds and products (milk and yoghurt) were screened for the presence of alkaloids, flavonoids, phenols, riboflavin and thiamine using analytical laboratory methods for basic screening, high-performance liquid chromatography (HPLC) and gas chromatography (GC) for quantification. The antimicrobial activity involved direct bioautography and minimum inhibitory concentration (MIC) against six antibiotic-resistant microorganisms, Acinetobacter baumannii ATCC 19606T, Enterococcus faecalis ATCC 29212, Klebsiella pneumoniae subsp. pneumoniae ATCC 700603, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus subsp. aureus ATCC 33591 and Candida albicans ATCC 24433. For the seed extracts, flavonoids and phenols were highly concentrated in the red and brown hulls of BGN compared to whole and dehulled BGN. Organic solvents in comparison to water yielded the highest concentration of flavonoids, whilst water yielded the highest concentration for phenols. Flavonoid compounds that were detected at the highest concentrations were rutin (24.458 ± 0.234 mg.g-1, brown hull extracted with 70% methanol), quercetin (0.070 ± 0.043 mg.g-1, red hull extracted with 70% methanol), kaempferol (0.391 ± 0.161 mg.g-1; brown hull extracted with 70% ethanol) and myricetin (1.800 ± 0.771 mg.g-1; red hull extracted with 70% methanol). For phenol compounds, gallic acid (0.009 ± 0.004 mg.g-1; brown hull extracted with milli-Q water), catechin (0.026 ± 0.041 mg.g-1; brown hull extracted with milli-Q water), methyl gallate (0.008 ± 0.013 mg.g-1; brown whole extracted with milli-Q water), chlorogenic acid (0.115 ± 0.199 mg.g-1; brown hull extracted with milli-Q water) and ellagic acid (0.105 ± 0.082 mg.g-1; red hull extracted with milli-Q water) were detected. Vitamins B1 and B2 (riboflavin and thiamine) were mostly present in milli-Q water extracts. Black-eye hull had the highest concentration of thiamine (vitamin B1) and riboflavin (vitamin B2) consisting of 0.072 mg.g-1 (extracted with milli-Q water) and 0.002 mg.g-1 (extracted with 70% ethanol and 70% methanol). Red and brown hull extracts from organic solvents (70% ethanol and 70% methanol) showed the highest antimicrobial activity, whereas the whole, dehulled and hulls (black-eye and brown-eye) extracts had no antimicrobial activity. As for BGN products extracts, flavonoid compounds that were detected at the highest concentrations were rutin (5.694 mg.g-1, whole BGN milk, milli-Q water), quercetin (0.703 mg.g-1, whole BGN yoghurt, milli-Q water) and myricetin (0.987 mg.g-1, whole BGN yoghurt, 70% ethanol).

Submitted in complete fulfillment for the degree of Doctor of Philosophy (Food Science and Technology), Durban University of Technology, Durban, South Africa, 2016. / Bambara groundnut (Vigna substerranea) is an indigenous legume of African origin which is currently experiencing a low level utilisation. It is tolerant to drought and can grow under poor soil conditions in which other lucrative crops such as groundnut cannot grow. Bambara is a good source of protein comparable to that of cowpea and slightly lower than soya bean. In order to assess the potential use of bambara protein as a functional ingredient in food systems and as an important ingredient for the formulation of therapeutic product, the knowledge of its protein composition, structure and functionality becomes important. The main goal of this thesis was to determine the composition and bioactive properties of bambara protein and its hydrolysates. Specifically, a comparative study was carried out on the protein content, yield and functional properties of protein concentrates prepared from three different bambara landraces using different extraction methods (Salt solubilisation and Acid precipitation). There was no significant difference in protein content, yield and functional properties of the landraces. However, the method of extraction had an influence on their physicochemical and functional properties. Acid precipitation produced bambara protein concentrates with high protein content and yield (79% and 52% respectively) when compared to salt solubilisation (protein content - 57% and yield - 25%). Protein concentrates prepared through salt solubilisation method exhibited better functional properties in terms of water absorption capacity, oil absorption capacity, foaming capacity, foaming stability and emulsion activities when compared to concentrates obtained through acid precipitation. Furthermore, the composition of bambara proteins produced through isoelectric precipitation was determined. SDS PAGE revealed four major bands; a broad band at 55 kDa which was analysed to be vicilin, two medium bands at 62 kDa and 80 kDa and a high molecular weight (HMW) protein at 141 kDa. Further investigation of bambara protein revealed vicilin (55 kDa) with two sub units as the major protein in bambara and this was also confirmed by the proteomic map. The proteomic map revealed acidic amino acids as the major protein of bambara which is characteristic of vicilin, the map also showed that there were differences in the number of spots across the landraces with 77 spots matching each other. Circular dichroism spectroscopy exhibited reductions in α-helix, and β-pleated sheet conformations as pH varies. In addition, the tertiary structures as observed from the near-UV CD spectra were also influenced by shifts in pH conditions. Differential scanning calorimetry thermograms showed two endothermic peaks at around 67 and 81oC respectively. These can be attributed to thermal denaturation of vicilin and the HMW protein. Subsequent studies used isolates from red bambara since the composition of the landraces were similar. Bambara protein isolate was subjected to enzymatic hydrolysis using three proteases (alcalase, pepsin and trypsin) to produce various bambara protein hydrolysates (BPHs). BPHs were investigated for antioxidant and antihypertensive activities. The in vitro structural and functional characteristics of bambara protein and its enzymatic protein hydrolysate revealed that bambara groundnut possessed antioxidant properties against a variety of physiologically relevant free radicals. High surface hydrophobicity and the molecular size of the peptide seem to be important for scavenging of hydroxyl radicals, ferric reducing power and metal chelation. BPHs and peptide fractions were able to scavenge DPPH radicals with greater affinity for smaller size. Less than 1 and 1-3 kDa pepsin fraction was able to scavenge DPPH radical more than glutathione, BPHs and its fractions scavenge ABTS•+ three folds than the isolate. Scavenging of superoxide radicals was generally weak except for 5-10 kDa peptide fractions. All BPHs inhibited linolenic acid oxidation with greater affinity for the lower molecular size peptide. BPHs showed potential antihypertensive properties because of the in vitro inhibition of activities of angiotensin converting enzyme (ACE) and renin inhibition. The molecular size had significant effect on the ACE inhibitory properties with low molecular weight peptide (<1 kDa) fractions exhibiting significantly higher (p<0.05) inhibitory activities. However, enzyme type had synergistic effects on renin inhibition with alcalase hydrolysate showing highest inhibition at 59% when compared to other hydrolysates and their membrane fractions. The fractions with <1 and 1-3 kDa peptides showed a higher potential as antihypertensive and antioxidant peptides. Based on this study, incorporation of bambara protein isolate as an ingredient may be useful for the manufacture of high quality food products. Likewise, the bambara protein hydrolysates, especially the <1 kDa and 1-3 kDa fraction represent a potential source of bioactive peptides in formulating functional foods and nutraceuticals. / D

Thesis (Master of Applied Sciences in Chemistry)--Cape Peninsula University of Technology, 2018. / Antimicrobial Peptides (AMPs) are short peptides of about 45 - 54 amino acids that exhibit antibacterial and antifungal activities. Plant defensin is a type of AMP in plants which belong to a family of cationic peptides with a characteristic 3D folding pattern held in place by four disulfide bridges. AMPs especially defensins have been identified to have a huge biotechnological potential and are being patented for many applications. The aim of this work was to clone an antimicrobial peptide from Vigna subterranea and characterise it with bioinformatics analysis. 4 sets of primers were synthesized according to the sequences of conserved regions in AMPs i.e. defensins from legumes like Vigna unguiculata, Vigna radiata, Cicer arietinum and Cajanus cajan, amongst others, which have defensins with only a few sequence differences. The primers were designated VsDef P1 to P4. Using Vigna subterranea total genomic DNA as a template, fragments of expected sizes were successfully amplified and cloned into the pDRIVE vector and used to transform Escherichia coli JM109 cells in each case. Representative clones were sequenced and analysed using BLAST from National Center for Biotechnology Information. However, only the VIG clone was shown to be a bona fide defensin (over 90% identity, E-value of 1ex102, 99% query coverage of the nucleotide sequence, compared to Vigna unguiculata defensin). Based on this high sequence identity, a new pair of primers VsDef P5 was designed based on the Vigna unguiculata defensin sequence to specifically amplify the complete Vigna subterranea defensin gene, hereafter called VsDef1. Attempts to clone VsDef1 were however unsuccessful, and evidence of clone deletion and insert re-arrangement of insert DNA was observed. Direct sequencing of the PCR product demonstrated that it was indeed the complete VsDef1 pre-protein, composed of 433 nucleotides. In silico translation and analysis showed that VsDef1 has an intron at position 105 − 259 of the nucleotide sequences and encodes for a 78 amino acid peptide. Phylogenetic analysis revealed to be similar to the sequence of the defensins for Vigna unguiculata (96%), Vigna radiata (95%), Vigna angularis (95%) and Phaseolus vulgaris (93%) on the NCBI database. The three - dimensional structure of the peptide was modelled with SWISS-MODEL expasy and the structure was found to include one α- and three β domains, similar to those of other defensins. The failure to identify VsDef1 clone in a V. subterranea library and the failure to recover its cDNA clone are consistent with the hypothesised toxicity of VsDef1 to Escherichia coli. It is suggested that a different host, such as yeast, should be used in the future. The VsDef1 mRNA levels in germinating V. subterranea seeds was however successfully investigated using real-time reverse transcription quantitative PCR. VsDef1 mRNA is present in both the testa and embryo of dry seed and will persist through the early stages of seedling growth. This demonstrates the importance of VsDef1 in fighting off infection during germination in order to ensure successful germination. It is therefore essential to characterise more antimicrobial peptides from V. subterranea. The diversity of AMPs and their patterns of expressed genes will enable understanding of complex regulatory networks, which will likely enable identifying of genes involved in diseases and new biological processes.