Pillai, Santhosh Kumar KuttanOkuofu, Somiame Itseme2021-07-292021-07-292020https://hdl.handle.net/10321/3607Submitted in fulfilment of the requirements for the Degree of Master of Applied Sciences in Biotechnology, Durban University of Technology, Durban, South Africa, 2020.Bambara and cowpea are important pulses grown in semi-arid South Africa due to their balanced nutrient profile and drought resilient capacity. The haulm is the lignocellulosic residue obtained after grain harvest and are rich in carbohydrates. However, these haulms are underutilised and under researched. The aim of the study, therefore, was to investigate the potential to valorise bambara haulms (BGH) and cowpea haulms (CH) to bioethanol which is the most promising biofuel with commercial prospects currently. The structural and chemical composition of BGH and CH was elucidated using techniques such as compositional analysis, XRD, FTIR, ICP-AES, and SEM. Results indicated a volatile matter and fixed carbon mass fraction of 77.70% and 13.15% (w/w) in BGH and 76.16% and 16.26% (w/w) in CH respectively. The polysaccharides make up the largest fraction (51%), followed by extractives (> 20%), while the lignin in BGH (12%) and CH (10%) was low. X-ray diffraction pattern showed a higher percentage of amorphous regions in BGH (78%) than CH (56%). CH was then subjected to dilute acid pretreatment (DAP) to enhance biosugar production for bioethanol fermentation. The effects of operational factors for DAP including temperature, time, and acid concentration on sugar yield and inhibitor formation was investigated and optimised using response surface methodology (RSM). The solid recovered after DAP was subjected to prehydrolysis with simultaneous saccharification and fermentation (PSSF). In addition, the pretreatment hydrolysate was detoxified and fermented to ethanol using cocultures of Saccharomyces cerevisiae BY4743 and Scheffersomyces stipitis wild type (PsY633). A total ethanol titre of 15.67 g/L was obtained corresponding to 75% conversion efficiency. On the other hand, BGH was subjected to deep eutectic solvent (DES) pretreatment. Five deep eutectic solvents were prepared and screened for their effectiveness in improving enzymatic sugar yield. This was achieved by pretreating BGH with each DES followed by a 48 h enzymatic saccharification. Choline chloride – lactic acid (ChCl-LA) treatment provided the most promising result and was further optimised by investigating the effect of different temperatures and time on cellulose loss and enzymatic sugar yield. ChCl-LA pretreatment at 100°C for 1 h was observed to be the best condition for maximum sugar recovery. The hydrolysate thus obtained was concentrated and fermented for 72 h with S. cerevisiae BY4743. A maximum ethanol yield of 11.57 g/L was obtained. From the results, it is evident that bambara and cowpea haulm are promising substrates for bioethanol production. Dilute acid hydrolysis was shown to be effective in the pretreatment of CH with over 85% of the theoretical sugar recoverable for conversion to bioethanol. In addition, deep eutectic solvents are effective media for breaking the recalcitrance in BGH to achieve high sugar yield for conversion to bioethanol. However, further studies are required to reduce cellulose loss during pretreatment to improve bioethanol yield.139 penBambara groundnutCowpeaHaulmBambara groundnutCowpeaLignocelluloseBiomass energyEnergy cropsThesishttps://doi.org/10.51415/10321/3607