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The ethanol yields from lignocellulo-starch biomass (peels of sweet potato, elephant foot yam, tannia, greater yam and beet root) by fed-batch separate hydrolysis and fermentation (F-SHF) and simultaneous saccharification and fermentation (F-SSF) using Saccharomyces cerevisiae were compared. Fed-batch saccharification of steam or dilute sulphuric acid pretreated biomass enhanced the reducing sugar yield which resulted in high RS consumption, volumetric ethanol productivity and ethanol yield during the first 24 h fermentation under F-SHF mode, while continuous production and utilization of reducing sugars occurred up to 72 h in F-SSF. Dilute sulphuric acid pretreated residues under F-SHF gave higher ethanol yield (34–43 g/L) and productivity (274–346 ml/kg dry biomass) than steam pretreatment (27–36 g/L and 223–295 ml/kg respectively), while F-SSF was superior for steam pretreated peels of sweet potato, elephant foot yam and tannia giving ethanol yields from 281 to 302 ml/kg. Glucose and xylose were present in all the hydrolysates with a preponderance of glucose and fermentation resulted in significant reduction in glucose levels in both F-SHF and F-SSF. Higher levels of total soluble phenolics and hydroxymethyl furfural were observed in the hydrolysates from dilute sulphuric acid pretreatment and yeast assimilated/detoxified part of the inhibitors, while only trivial amounts of furfural were present due to the low xylose content in the hydrolysates. Continuous formation led to higher accumulation of inhibitors in F-SSF despite supplementation with the detoxification mix comprising Tween 20, polyethylene glycol and sodium borohydride. F-SHF of dilute sulphuric acid pretreated biomass could be considered as a comparatively advantageous process where only one time feeding of enzyme cocktail and yeast was adopted compared to multiple feeds of enzymes and yeast along with other additives such as detoxification mix or nutrient solution in F-SSF.

The generation of phenolic inhibitors in the prehydrolysates from steam, dilute sulphuric acid (DSA) and lime pretreated root wastes (peels from sweet potato, elephant foot yam, tannia, beet root and greater yam) and vegetable processing residues (peels from ash gourd, pumpkin and vegetable banana and mixed vegetable waste) was monitored. Further, the effect of detoxification agents such as Tween 20, PEG 4000, active charcoal and sodium borohydride in reducing their levels was compared. The aqueous extracts from native biomass contained soluble phenolics at levels of 1.03% to 3.02%. Highest release (12.5-16.2% increase from the values in aqueous extracts of native biomass) of total soluble phenolics (TSPs) into the prehydrolysates was found in steam (60 min.) and DSA (60 min; 121 ⁰C) pretreatments, irrespective of the type of biomass, while lime pretreatment resulted in only 1.7-3.1% increase in the phenolics. Maximum quantity of TSPs was removed in 2 h at room temperature (30±1 ⁰C) by Tween 20 (70-81%) or its combination with PEG (73-82%). Active charcoal (2%; 45 ⁰C; 1 h) or sodium borohydride (40 mM; 20 min; 30±1 ⁰C) removed only 40-59% and 37-53% phenolics respectively. Loss of reducing sugars was the least (2-3%) in Tween 20 (0.50%) treatment, indicating its superiority over the other treatments.

Two strategies leading to enzyme saving during saccharification of pretreated lignocellulo-starch biomass (LCSB) was investigated which included reducing enzyme dosage by varying their levels in enzyme cocktails and enhancing the fermentable sugar yield in enzyme-reduced systems using detoxification chemicals. Time course release of reducing sugars (RS) during 24–120 h was significantly higher when an enzyme cocktail containing full dose of cellulase (16 FPU/g cellulose) along with half dose each of xylanase (1.5 mg protein/g hemicelluloses) and Stargen (12.5 μl/g biomass) was used to saccharify conventional dilute sulphuric acid (DSA) pretreated biomass compared to a parallel system where only one-fourth the dose of the latter two enzymes was used. The reduction in RS content in the 120 h saccharified mash to the extent of 3–4 g/L compared to the system saccharified with full complement of the three enzymes could be overcome considerably by supplementing the system (half dose of two enzymes) with detoxification chemical mix incorporating Tween 20, PEG 4000 and sodium borohydride. Microwave (MW)-assisted DSA pretreated biomass on saccharification with enzyme cocktail having full dose of cellulase and half dose of Stargen along with detoxification chemicals gave significantly higher RS yield than DSA pretreated system saccharified using three enzymes. The study showed that xylanase could be eliminated during saccharification of MW-assisted DSA pretreated biomass without affecting RS yield when detoxification chemicals were also supplemented. The Saccharification Efficiency and Overall Conversion Efficiency were also high for the MW-assisted DSA pretreated biomass. Since whole slurry saccharifcation of pretreated biomass is essential to conserve fermentable sugars in LCSB saccharification, detoxification of soluble inhibitors is equally important as channelling out of insoluble lignin remaining in the residue. As one of the major factors contributing to the cost of ethanol production from LCSB is the cost of enzymes, appropriate modification of enzyme cocktail based on the composition of the pretreated biomass coupled with effective detoxification of the slurry would be a promising approach towards cost reduction.

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