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Natural and Lactic Acid Bacteria Fermentations of Pastes of Soybeans and Soybean-Maize Blends

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Natural and Lactic Acid Bacteria Fermentations of Pastes of Soybeans and Soybean-Maize Blends Natural and Lactic Acid Bacteria Fermentations of Pastes of Soybeans and Soybean-Maize Blends: Effect on Nutritional Quality, Microbial iversity, Food Safety and Consumer Acceptance Naturlig fermentering og fermentering med melkesyrebakterier av soyabnne- og soyabnne-mais blandinger: Effekt p ernringsmessig kvalitet, mikrobiell diversitet, mattrygghet og forbrukeraksept Philosophiae Do tor (PhD) Thesis Tinna Austen Ng&ong&ola-Manani Department of Chemistry, Biote hnology and Food S ien e Fa ulty of (eterinary Medi ine and Bios ien es Norwegian University of ,ife S ien es -s .011 Thesis number .011:21 ISSN: 1241-510. ISBN: 472-2.-777-1.18-4 Table of Contents !/Ybh[95D9a9b 595L/! Lhb LL !..w9L! Lhb LLL aa!w L !aa9b5w!D LL [L hC t!t9w Lb wh5/ Lhb .!/YDwh b5 h I9 5 !9a9b hC I9 twh.[9a W LCL/!Lhb hC I9 5 [L 9w! w9 w9L9 ILhw twh5 /Lhb !b5 9 hC h.9!b /I9aL/![ !b5 b wLLhb![ /hathLLhb hC h.9!b 9CC9/ hC 5ha9L/ twh/9LbD hb /hathLLhb hC h.9!b C9wa9b!Lhb ! ! a![[ /![9 !b5 Ih 9Ih[5 9/Ibh[hD aL/wh.Lh[hD hC [!/L/ !/L5 .!/9wL! C9wa9b95 /9w9![ Dw 9[ hC !CwL/! aL/wh.Lh[hD hC ![Y![Lb9 C9wa9b95 995 !b5 h.9!b twh5 / ! ! !C9 hC C9wa9b95 h.9!b twh5 / " a9Ih5 Chw 599/Lhb hC aL/whhwD!bLa Lb C9wa9b95 h.9!b twh5 / # 9bhw twht9wL9 !b5 !//9t!b/9 hC C9wa9b95 h.9!b twh5 / $ a9!.h[La hC I9 a!Whw aL/whhwD!bLa Lb%h[%95 Lb C9wa9b!Lhb hC h.9!b t!9 " h.W9/ L9 a!Lb h.W9/L%9 t9/LCL/ h.W9/L%9 IthI99 a!Lb w9 [ !b5 D9b9w![ 5L / Lhb 9CC9/ hC C9wa9b!Lhb hb b wLLhb![ v ![L hC t!9 hC h.9!b !b5 h.9!b a!L'9 .[9b5 ! aL/wh.L![ tht [!Lhb 5L%9wL !b5 a9!.h[La Lb b! w![ !b5 [!/L/ !/L5 .!/9wL! C9wa9b!Lhb # Chh5 !C9 Lat[L/!Lhb hC b! w![ !b5 [!/L/ !/L5 .!/9wL! C9wa9b!Lhb 9bhw twht9wL9 5wL%LbD /hb a9w !//9t!b/9 ! /hb/[ Lhb !b5 C w9 t9w t9/ L9 ! /hb/[ Lhb $ C w9 t9wt9/L%9 !tt9b5L/9 w9C9w9b/9 " 9b/[h 95 t!t9w L#L " ( ACKNOWLEDGEMENTS It is with gratitude that I acknowledge the Norwegian Programme for Development, Research and Education (NUFU) for financing this research. The International Foundation for Science (IFS) through grant number E/4889-1 is acknowledged for financing some of the research activities conducted in Malawi. I also acknowledge the Norwegian State Education Loan Fund (Lånekassen) for financially supporting my stay in Norway. I sincerely thank management of Lilongwe University of Agriculture and Natural Resources (LUANAR), Bunda Campus, Malawi for granting me a study leave. I am very grateful to the coordinators of the Lungwena health, nutrition and agriculture research project (NUFUPRO-2007/10149), Professor Kenneth Maleta and Professor Johanne Sundby for accepting me as a PhD student. I sincerely thank Dr Anne Wetlesen for accepting to be my supervisor when I was applying for admission at Norwegian University of Life Sciences (NMBU). Dr Charles Masangano is also thanked for coordinating at LUANAR. This PhD thesis would not have been accomplished if it were not for the guidance and support from my supervisors. I thank all my supervisors for reading my papers over and over. I am forever grateful to Associate Professor Trude Wicklund for accepting to be my main supervisor when Dr Anne Wetlesen took another job. Associate Professor Trude Wicklund was supportive and considerate in all matters. I am deeply indebted to my co-supervisor Associate Professor Hilde Marit Østlie for her keen interest in my work. Her in-depth microbiological knowledge and her timely constructive suggestions shaped the write-up of the papers for the better. I also greatly appreciate the kindness and generosity that Associate Professor Hilde Marit Østlie and her family accorded my family when we just moved to Norway. I also express my appreciation to my co-supervisor Dr Agnes Mwangwela for her valuable contribution towards this work and because of her administrative support at LUANAR that created a conducive environment for laboratory work in Malawi. I am very grateful for the technical support I received from members of the Dairy Technology group (1951) at IKBM, Norway. I extend my gratitude to Kari Olsen for HPLC analyses, to Ellen Skuterud for texture analyses, to Tone Stokke Molland for assistance in some microbiological techniques, and to May Aalberg for countless everyday support in the lab. I also appreciate the assistance of Ahmed Abdelghani and Bjørg Holter in the pathogen lab. I am thankful for the technical support I received in Malawi from the following: Mr Chipiliro Kantikana (Home Economic and Human Nutrition Department) and Mr Emmanuel Mbewe ( (Crops and Soil Sciences Department). Sincere gratitude goes to Oliveta Buleya for practical support throughout laboratory analyses and for protein analyses. Technicians in Aquaculture and Fisheries Sciences and Basic Sciences departments are thanked for their support in various ways. I am very thankful to the students of Home Economics and Human Nutrition Department and to the staff members of LUANAR, Bunda Campus, who participated in descriptive sensory panel. The people of Lungwena (Mdala-Makumba, Chilonga and Chapola villages) who participated in consumer studies are also thanked. I greatly thank Mrs Mercy Chautsi for tirelessly supporting me during sensory and consumer studies. I greatly appreciate former PhD students of IKBM. Special thanks to Dr Beatrice Mgaya Kilima for supporting me when I needed someone most, the words of encouragement and the laughter we shared are cherished. I am very grateful to Dr Agnes Nabubuya for sharing in the joys and the challenges. I thank Dr Heide Grønnevik for hands-on DGGE tutorial and Dr Ivan Mukisa for fruitful discussions on molecular microbiology work. I am thankful for the wonderful association with the Malawian community in Ås and around Oslo. Thanks to Doreen Auma and Cecilia Munthali for their social support. Many and special thanks go to my Mum and Dad for laying good foundation, encouraging and supporting their children with education from primary school to universities. Your love, prayers, guidance and innumerable support are always cherished. I sincerely appreciate the love, support, encouragement and prayers from my sisters (and their families) Anita, Esther, Mafase and Khumbo, my brother Austin, my Aunt Mrs Deliwe Munthali and my niece Maggie Munthali. My sincere and heartfelt gratitude goes to my dear husband Felton for supporting me throughout my studies. I thank Felton for taking good care of our son, Takondwa when I was away. I also thank my beloved children Takondwa and Mphatso for their patience when I was stressed and gave them divided attention. Your smiles and joys are a motivation. Lastly but not least, I praise and thank God for giving me life and for granting me rare opportunities and blessings in life. Tinna Austen Ng’ong’ola-Manani. Ås, Norway, September, 2014. ( DEDICATION To My Dad, Austin Henderson Supuni Ng’ong’ola and my mum, Grace Chapasi Ng’ong’ola . For your love and for instilling love for education in your five daughters and your son. & To Felton, Takondwa and Mphatso . For your love, patience and endurance during my studies. ( ABBREVIATIONS 100S Paste composed of 100% soybeans and subjected to natural fermentation 100SBS Paste composed of 100% soybeans and subjected to lactic acid bacteria fermentation through back-slopping 100SC Paste composed of 100% soybeans and fermented with Lactobacillus fermentum 75S Paste composed of 75% soybeans and 25% maize and subjected to natural fermentation 75SBS Paste composed of 75% soybeans and 25% maize and subjected to lactic acid bacteria fermentation through back-slopping 90S Paste composed of 90% soybeans and 10% maize and subjected to natural fermentation 90SBS Paste composed of 90% soybeans and 10% maize and subjected to lactic acid bacteria fermentation through back-slopping 90SC Paste composed of 90% soybeans and 10% maize and fermented with Lactobacillus fermentum BSP Pastes fermented through back-slopping CFU Colony forming units DGGE Denaturing gradient gel electrophoresis DNA Deoxyribonucleic acid LAB Lactic acid bacteria LFP Lactic acid bacteria fermented pastes NFP Naturally fermented pastes PCA Principal component analysis PCR Polymerase chain reaction SCP Pastes fermented with Lactobacillus fermentum as starter culture TI Trypsin inhibitors TIA Trypsin inhibitor activities ( SUMMARY Traditional Malawian diets are predominantly maize-based and have been associated with widespread inadequate intakes of several nutrients. In addition to maize, legumes are an important source of protein and other nutrients in diets of many people in developing countries. Soybeans have the highest protein content among legumes and when consumed together with cereals, a high quality protein is provided because cereals and legumes are complementary in terms of limiting amino acids. However, soybean utilization in Malawi is minimal due to limited knowledge in processing. In an effort to increase utilization and consumption of soybeans by all age groups in Malawi, solid-state fermented pastes of soybeans and soybean-maize blends were developed. The fermented pastes were to be used as relish and to serve as major sources of protein in maize- based diets. Spontaneous solid state fermentation of soybeans favors growth of Bacillus subtilis , a highly proteolytic organism that produces high amount of ammonia. High ammonia levels result in strong odor which some people find objectionable. On the other hand, lactic acid bacteria (LAB) are weakly proteolytic and do not lead to accumulation of high levels of organoleptically unpleasant metabolic products. In this study, thobwa , a Malawian fermented cereal gruel prepared from maize flour and co- fermented with malt flour from finger millet was used as a back – slopping material to facilitate lactic acid bacteria fermentations in LAB fermented pastes (LFP).
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