Enzymatic hydrolysis at high solids loading has the potential to reduce both capital and operational expenditures. Here, pretreatment of rice straw (PRS) with dilute acid was carried out at a pilot ...scale (250 kg per day) at 162°C for 10 min and 0.35% acid concentration, followed by enzymatic hydrolysis at different total solids loadings. It showed that although the total sugar concentration increased from 48 to 132 g/l, glucan conversion reduced by 27% (84–66.2%) with increasing solids from 5 to 20% in batch mode. Therefore, two different fed-batch approaches were evaluated to improve the glucan conversion by the sequential addition of a substrate and/or enzyme. At 20% solid loadings and a 3 filter paper units/g enzyme dosage, the highest glucan conversion obtained was 66% after 30 h of hydrolysis in batch mode. However, in an optimized fed-batch approach, the glucan yield was improved to 70% by simply dividing the substrate feeding into three batches, that is, 50% at 0 h, 25% each after 4 h, and 8 h of hydrolysis reaction. The addition of surfactant (Ecosurf E6) further improved the conversion to 72% after 30 h. The role of critical factors, that is, inhibitors, enzyme–lignin binding, and viscosity, was investigated during the course of hydrolysis in the batch and fed-batch approaches. This study suggests a sustainable approach for improved hydrolysis at high solids loadings by fine-tuning a simple process.
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•Pilot-scale pretreatment of rice straw in a 250 Kg/day plant capacity.•26% reduction in glucan conversion at high solids loading of 20% w/w.•Enzyme inhibition at high solid loading ...due to feedback inhibition, lignin binding.•Six strategies investigated to enhance the glucan conversion yields.•15% increase in glucan conversion by enzyme blending approach.•A unique strategy developed showing 24.7% enhancements in biomass saccharification.
The commercial viability of 2G bioethanol production is dependent on cost-effective and efficient enzymatic hydrolysis of biomass at low enzyme and high dry solid loadings. In the present study, rice straw was subjected to a dilute acid pretreatment in a continuous pilot plant (250 kg/day).It was observed that, the glucan conversion yields were reduced by 26% at high solid loading (20% w/w) with low enzyme dosage of 3 FPU/g w/w of in-house developed enzyme (MRJ16). Thus, multiple approaches were investigated to enhance the glucan conversion yields, a) fed batch approach (splitting biomass or enzyme dosage during process), b) additives supplementation (surfactants, metal ions, non-enzymatic proteins and activated charcoal) and c) enzyme blending with accessory enzymes. Results showed that, the 3-stage fed-batch approach enhanced the hydrolysis yield by 12% in comparison to 14.3% increase with the selected surfactant (0.5% w/w Ecosurf). The selected metal salt (NiCl2, 1 mM) increased the glucan conversion by 13.2%, while, the non-enzymatic proteins (5% w/w yeast hydrolysate and yeast extract) improved the yields to 14.8% and 14.4%, respectively. The activated charcoal at a concentration of 0.5% w/w enhanced the hydrolysis yields by 14.4%. Further, 15.1% increase in sugar yield was obtained by supplementing the in-house produced enzyme (MRJ16) with xylanase from Thermomyces lanuginosus and BGL from Penicillium janthinellum EMS-UV-8. Finally, a unique optimized strategy was developed on the basis of selected approaches and parameters which demonstrated 24.7% higher saccharification in comparison to control. To the best of our knowledge, this is the only study available in literature where; a wide range of strategies were studies on a single feedstock (i.e. rice straw) pretreated at pilot-scale and determine its effect for improved enzymatic hydrolysis. Furthermore, developed process helps us to save (almost 2X) expensive enzyme to achieve same level of glucan conversion (∼62.0%) to make the overall process cost effective by using cheap and easily available additives. The current study also provided an in-depth understanding of enzyme inhibition at low enzyme dosages and high solid loadings in an industrially relevant scenario.
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