Although sorghum malt is relatively rich in free amino nitrogen (FAN), the 150 mg FAN/L threshold recommended for brewing is difficult to obtain. The vitreous nature of the sorghum endosperm hinders proteolysis during brewing. Hence, exogenous proteolytic enzymes are often required to increase hydrolysis of sorghum malt protein to produce sufficient FAN in order to support rapid yeast growth during fermentation. Ten exogenous proteases were examined for their production of FAN in sorghum malt mashing. Mashing was done at 550C for 45 minutes. Levels of FAN, as determined by the ninhydrin method, showed great variation among the proteolytic enzymes, ranging from 96 in control to 182 mg/100 g malt with possibly of most effective proteolytic enzyme. The variation in FAN level was possibly due to different optimal mashing conditions of exogenous proteases used and perhaps due to low ratios of exopeptidase/endopeptidase in the enzyme preparations. Low temperature (400C) and long duration mashing for (7 hours) gave good FAN production during mashing to a total of 113 and 138 mg/100 g malt in control and the treatment with exogenous proteolytic enzyme Flavourzyme plus malt, respectively. The exogenous enzyme (Flavourzyme) plus potassium metabisulphite (PMB) increased FAN production during mashing in the ratio of 2 to 1 in a treatment where PMB was added compared to one without. Similarly, hot wort extract (HWE) increased by 8% during mashing with exogenous enzyme plus PMB compared to one without PMB, respectively. PMB was involved in destabilizing the disulphide bonds in the sorghum protein polypeptide chains allowing proteolytic enzymes better accessibility to proteins. The increase in HWE was possibly due to the starch being freed from the sorghum protein matrix. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed some oligomeric and polymeric kafirins after mashing. With transmission electron microcopy (TEM), protein bodies of varying sizes with partially degraded peripheral edges and some holes were seen after mashing. SDS-PAGE and TEM results suggest insufficient proteolysis. High protein digestibility sorghum’s potential for brewing was examined with reference to FAN production. Although during mashing FAN increased by approx. 82 and 115% for unmalted normal and high digestibility sorghums, respectively, the 150 mg FAN/L threshold, recommended for brewing was not achieved. FAN production to levels above 150 mg/L may only be realized if normal sorghum malt or high protein digestibility sorghum malt is mashed with exogenous enzymes containing sufficient exopeptidases coupled with appropriate mashing conditions. Copyright / Dissertation (MSc)--University of Pretoria, 2009. / Food Science / unrestricted

Master of Science / Food Science Institute, Grain Science and Industry / Jeffrey A. Gwirtz / There is a limited market of gluten-free beer for the 1% of the US population that is diagnosed with an autoimmune response to gluten protein known as celiac disease. Sorghum can be malted and used in the brewing process to replace malted barley, a grain toxic to celiac patients. The objective of the study was to develop an optimum brewing procedure for a gluten-free ale-style beer. Four different sorghum hybrids (82G63, 83G66, RN315, and X303) were malted and used in brewing gluten-free ale and evaluated for physical and chemical property differences. The four sorghum hybrids were characterized first as grain and then as malt using proximate analysis, single kernel characterization system (SKCS), amylose, α-amylase, and β-amylase contents. Isolated starch from unmalted and malted samples was evaluated with differential scanning calorimetry (DSC). Malt was evaluated throughout the malting process and percent nitrogen, percent moisture, 72 hr germinative energy, steep out moisture, germination-end, and malting loss were measured. Malted sorghum hybrid samples were milled into grist, and employed in a double mash, double decoction brewing process. Following the brewing process the wort was evaluated for specific gravity, Brix, pH, color and free α-amino nitrogen (FAN). Wort was also analyzed using HPLC for ethanol and glucose content. The fermented beer was analyzed for specific gravity, Brix, pH, alcohol by volume, and color. HPLC was also used to measure ethanol and glucose content. Results of analysis found that a significant difference (p=0.05) was found for the DSC data onset temperature, which ranged from 61.75 to 65.51, illustrating the difference in starch gelatinization temperature compared to other cereals. A significant difference was found in α-amylase content (p=0.05) which ranged from 0.16 to 058 in unmalted sorghum and 71.63 to 96.44 in malted sorghum. In addition, α-amylase and β-amylase contents increased during malting. HPLC analysis of wort indicated a significant difference was found in percent maltose which ranged from 1.27 to 2.81. FAN content of wort was also significantly different and ranged from 65.15 to 151.37. HPLC of beer showed a significant difference in percent ethanol and percent glucose. Percent ethanol in the final beer ranged from 3.28 to 4.17 and percent glucose range from 0.16 to 0.31. Process development evaluation indicated a gluten-free ale style beer could be successfully produced with 100% sorghum malt.

Due to the high cost of wheat importation in countries where the climatic conditions do not favour its cultivation, alternative sources of bread baking flour are required. Maize is a suitable alternative because it is by far the most important crop produced in Africa. However, it lacks gluten, the protein that is formed in wheat dough which is responsible for the desirable quality attributes (high loaf volume, soft and open crumb structure) of wheat bread. Therefore the need arises to improve maize bread quality. The effects of three types of non-wheat bread methods on the quality of maize bread were investigated. The first was a traditional sourdough method used in Lesotho for making steamed bread. This involved addition of spontaneously fermenting sorghum malt sourdough (equivalent to 15% of the total maize flour) and pre-gelatinization of the starch in the maize flour with boiling water. The second was a Food and Agriculture Organization method which involved pre-gelatinization of the starch in 10% of the maize flour by cooking. The third method was a modern gluten-free sourdough method which involved fermenting 75% of the maize flour with a multiple strains starter culture or Lactobacillus plantarum plus the natural flora in the maize. The modern sourdough method produced maize bread with a more open crumb structure and a significant increase in loaf volume compared to the other methods. This was probably related to the high percentage of fermented maize flour in the recipe, which was probably sufficient to modify the dough properties satisfactorily enough to impact positively on the maize bread quality. Based on these findings, the modern sourdough method was investigated further. Maize sourdoughs were prepared (as described) and compared to chemically acidified maize dough. Sourdough maize bread had an approx. 25-26% increase in loaf volume and a more open crumb structure with large gas cells. This showed that the maize bread quality improvement was not due to low pH. Confocal laser scanning microscopy revealed a cohesive dough structure in the sourdoughs. Larger cells and a more uniform crumb structure were also observed in maize breads with maize sourdough. This indicated an improvement in the maize dough properties with sourdough. Differential scanning calorimetry showed that maize sourdough had a slightly lower peak temperature than straight maize dough, an indication of starch modification. Rheological analysis showed that maize sourdough had a shorter relaxation time, an indication that it was less elastic. Strain sweep analysis revealed that maize sourdoughs had the lowest elastic modulus, also indicating a less elastic dough. Temperature sweep analysis showed an initial less elastic dough and a final high tan delta, suggesting that the maize dough could withstand gas expansion pressure during baking without crumbling. The dominant lactic acid bacteria in the sourdoughs were identified as L. plantarum. In the two sourdoughs, the L. plantarum present were gram-positive, catalase negative and exhibited proteolytic activities. However, only the L. plantarum in the multiple strains starter culture fermented maize sourdough exhibited amylolytic activities. It is proposed that proteolytic activity of the L. plantarum degraded the endosperm protein matrix and hydrolysed the proteins soluble in the dough liquid, thereby allowing increased accessibility of water to the starch granules. It is further proposed that the amylolytic activity of the L. plantarum slightly hydrolysed the starch granules, increasing water absorption by the starch granules. It is proposed that improvement in maize bread quality by sourdough fermentation is due to starch modification (increase water accessibility and water absorption by the starch granules due to the proteolytic and amylolytic activities of the dominant lactic acid bacteria in the sourdoughs) which made the dough less elastic. This in-turn improves the ability of the dough to trap and withstand the pressure of the expanding carbon dioxide in the fermenting dough and bread. / Thesis (PhD)--University of Pretoria, 2014. / gm2015 / Food Science / Unrestricted