Maximum Glucose Yield Research Articles

Bioethanol Production from Ball-Milled and Regenerated Rice Husk

This research focused on the bioethanol production from rice husk via enzymatic hydrolysis and fermentation of glucose product. Rice husks were ball-milled, alkali pretreated with 2 %(w/v) NaOH for 3 hours and regenerated by N-methylmorpholine N-oxide (NMMO)/H2O solution into Cellulose II structure. These processes were aimed to enhance the surface purity area and accessibility of enzyme, respectively. Rice husks were hydrolyzed with cellulase from Trichodermareesei ATCC 26921 at various concentrations ranging from 10 to 40 FPU/g of rice husk. The maximum yields of glucose from alkali pretreated rice husk, ball-milled rice husk of sieve size 45-75 μm and regenerated rice husk were 7.13, 9.52 and 10.36 mg/mL, respectively. This was conducted at the enzyme concentration of 20 FPU/g of rice husk for 48 hours. The glucose product was fermented by using Saccharomyces cerevisiae Safale US-05 at various concentrations ranging from 10 to 70 %(v/v). The maximum yields of ethanol products at 50 %(v/v) of yeast and 48 hours of fermentation from the alkali pretreated rice husk, ball-milled rice husk of sieve size 45-75 μm and regenerated rice husk were 44.36%, 68.79% and 89.11% of glucose respectively. The greater yield may be arise from an increase in surface area and the swollen cellulose structure of rice husk.

Effects of reduced severity ammonia pretreatment on pelleted corn stover

Biomass densification impacts pretreatment efficacy and subsequent biochemical conversion to biofuels and other biobased chemicals. Pelleted corn stover was used to evaluate the soaking in aqueous ammonia (SAA) pretreatment efficacy at high solid loadings with reductions ammonia concentration, temperature, and pretreatment time. Pretreatment resulted in 70% greater delignification of stover pellets than loose stover. Glucose yields from enzymatic hydrolysis were 49% higher for the pelleted stover and yields did not change with a twofold increase in pretreatment solid loadings. Hydrolysis yields were modeled as a function of pretreatment conditions and the developed model predicted a maximum 24-h hydrolysis glucose yield of 96% at a pretreatment temperature of 60 °C, for 4 h, with 18% ammonia. Pretreatment severity can still be reduced while maintaining 90% or higher yields with different combinations of temperature, time, and ammonia concentration. Temperature was the most important pretreatment parameter within the design space for achieving high glucose yields. FTIR and SEM analysis of SAA-pretreated corn stover pellets illustrated that the pelleting process increases pretreatment efficacy by modifying the biomass structure, disrupting the lignin-hemicellulose linkages, and partially removing lignin. Using pelleted biomass can reduce the required severity of SAA-pretreatment while still producing glucose yields above 90%. Using pelleted corn stover as a biorefinery feedstock has potential to lower pretreatment costs in addition to the improved handling and transportation.

Multi-step approach to add value to corncob: Production of biomass-degrading enzymes, lignin and fermentable sugars

This work presents an integrated and multi-step approach for the recovery and/or application of the lignocellulosic fractions from corncob in the production of high value added compounds as xylo-oligosaccharides, enzymes, fermentable sugars, and lignin in terms of biorefinery concept. For that, liquid hot water followed by enzymatic hydrolysis were used. Liquid hot water was performed using different residence times (10–50min) and holding temperature (180–200°C), corresponding to severities (log(R0)) of 3.36–4.64. The most severe conditions showed higher xylo-oligosaccharides extraction (maximum of 93%) into the hydrolysates and higher recovery of cellulose on pretreated solids (maximum of 65%). Subsequently, hydrolysates and solids were used in the production of xylanases and cellulases, respectively, as well as, pretreated solids were also subjected to enzymatic hydrolysis for the recovery of lignin and fermentable sugars from cellulose. Maximum glucose yield (100%) was achieved for solids pretreated at log(R0) of 4.42 and 5% solid loading.

Glucose and Valuable Chemicals Production from Cotton Waste Using Hydrothermal Method

Direct hydrolysis of a towel to glucose was investigated using steam explosion and microwave-assisted treatment to find effective uses for cotton waste. The maximum glucose yield by direct hydrolysis (based on an untreated towel) was 18.8%, obtained at a steam pressure of 5.5 MPa (at 271 °C) and steaming time of 5 min using steam explosion. For the microwave-assisted treatment, with a 1.0 (w/w)% sulfuric acid catalyst, the maximum glucose yield by direct hydrolysis was 28.9%, obtained at a microwave heating temperature of 200 °C for 7 min. The maximum total glucose yield (from both direct hydrolysis and enzymatic hydrolysis of treated residue) was 78.0%, attained at a microwave heating temperature of 200 °C for 7 min with 0.5 (w/w)% sulfuric acid catalyst. Furthermore, the maximum total glucose and valuable water soluble chemicals (cellobiose, 5-hydroxymethylfurfural, formic acid, and levulinic acid), 94.1%, were achieved at heating temperature of 200 °C for 10 min with 0.5 (w/w)% sulfuric acid catalyst. Finally, ethanol, 84.5% of conversion rate, could be produced using supernatant (it contained glucose) and microwave treated residue (200 °C for 7 min with 0.25 (w/w)% sulfuric acid catalyst) as carbon source for Saccharomyces cerevisiae with less fermentation inhibition.

Effect of combined inorganic with organic acids pretreatment of rice straw on its structure properties and enzymatic hydrolysis

Rice straw was pretreated with sulfuric acid combined with five organic acids (oxalic acid, acetic acid, tartaric acid, succinic acid, and fumaric acid) at 121°C for 90 min with five different blending ratios. The concentrations of glucose, xylose, arabinose and inhibitors such as acetic acid, furfural and HMF (5‐hydroxymethylfurfural) in the prehydrolysate were analyzed. It demonstrated that the concentrations of both sugars and inhibitors in prehydrolysate decreased with increasing organic acids blending ratio. The different solid residues from pretreatments were subjected to enzymatic hydrolysis process. The results showed that pretreating rice straw using combined sulfuric acid with oxalic acid or fumaric acid resulted in the maximum glucose yield (∼66%) after 72 h enzymatic hydrolysis in sharp contrast to ∼58% glucose yield obtained from pretreatment only with sulfuric acid under the same conditions. Furthermore, the structure of the raw material and the pretreated rice straw were characterized by scanning electron microscopy (SEM) and Fourier transformer infrared (FTIR) spectrum to clarify the effects of the combined sulfuric acid with oxalic or fumaric acid pretreatment on disrupting the fibers, removing hemicellulose and making the cellulose structure loose. © 2017 American Institute of Chemical Engineers Environ Prog, 37: 808–814, 2018

Studies on the effect of hydrothermal pretreatment of sugarcane bagasse for biobutanol production

ABSTRACTThis study deals with the production of butanol using hydrothermally pretreated sugarcane bagasse (SCB) hydrolysate by anaerobic fermentation using Clostridium sporogenes. First, SCB was soaked in water at 30–70°C at a solid to liquid ratio (SLR) of 1:10. Second, SCB was soaked in water and subsequently subjected to thermal treatment at 121°C for 15 min. The effectiveness of pretreatment was evaluated by enzymatic hydrolysis and a maximum glucose yield of 462 mg/g of glucan was obtained from the pretreated biomass. Anaerobic fermentation of the hydrolysate under strict anaerobic condition resulted in the butanol yield of 0.173 g/g. C. sporogenes coculturing with Bacillus subtilis without anaerobic treatment was also investigated.

Enzymatic Hydrolysis of Lignocellulose for Bioethanol Production

AbstractThe effect of temperature, time and amount of enzyme on hydrolysis of wheat straw lignocellulose remaining after furfural production was studied. The residual substrate was subjected to enzymatic hydrolysis at different temperatures — 45 °C, 50 °C and 55 °C. Hydrolysis time was 72 hours, and samples were taken every 24 hours. The maximum glucose yield (76.5% of the theoretically possible) was reached when hydrolysis temperature 50 °C was used. The production rate of glucose increased with a hydrolysis period of time. The yield of glucose significantly depended on the ratio of enzyme to substrate.

Pre-treatment of biomasses using magnetised sulfonic acid catalysts

There is a significant interest in employing solid acid catalysts for pre-treatment of biomasses for subsequent hydrolysis into sugars, because solid acid catalysts facilitate reusability, high activity, and easier separation. Hence the present research investigated pretreatment of four lignocellulosic biomasses, namely Switchgrass (Panicum virgatum L ‘Alamo’), Gamagrass (Tripsacum dactyloides), Miscanthus (Miscanthus × giganteus) and Triticale hay (Triticale hexaploide Lart.) at 90°C for 2 h using three carbon-supported sulfonic acid catalysts. The catalysts were synthesized via impregnating p-Toluenesulfonic acid on carbon (regular) and further impregnated with iron nitrate via two methods to obtain magnetic A and magnetic B catalysts. When tested as pre-treatment agents, a maximum total lignin reduction of 17.73±0.63% was observed for Triticale hay treated with magnetic A catalyst. Furthermore, maximum glucose yield after enzymatic hydrolysis was observed to be 203.47±5.09 mg g–1 (conversion of 65.07±1.63%) from Switchgrass treated with magnetic A catalyst. When reusability of magnetised catalysts were tested, it was observed that magnetic A catalyst was consistent for Gamagrass, Miscanthus × Giganteus and Triticale hay, while magnetic B catalyst was found to maintain consistent yield for switchgrass feedstock. Our results suggested that magnetised solid acid catalyst could pre-treat various biomass stocks and also can potentially reduce the use of harsh chemicals and make bioenergy processes environment friendly.

Optimization of liquid ammonia pretreatment variables for maximum enzymatic hydrolysis yield of energy cane bagasse

This study aimed at optimizing a liquid ammonium hydroxide pretreatment for energy cane bagasse for maximum sugar yields (glucose and xylose) via response surface methodology. Optimized pretreatment parameters included temperature (160–220°C), ammonium hydroxide to biomass ratio (0–0.5:1) and residence time (30–60min). Temperature was found to be the dominant pretreatment parameter followed by ammonium hydroxide to biomass ratio. High temperatures and long residence times had a negative effect on sugar yields. Sugar yields had the highest correlation with lignin removal. Based on our quadratic models fitted on the experimental results, optimum pretreatment conditions for maximum glucose yield were 208°C, for 36min and at an ammonium hydroxide to biomass ratio of 0.4:1. A glucose yield of 30.77g glucose/100g (dry weight) untreated biomass and a xylose yield of 3.99g xylose/100g (dry weight) untreated biomass were predicted by the model. Optimum pretreatment conditions for maximum xylose yield were 160°C, for 60min and at an ammonium hydroxide to biomass ratio of 0.31:1. These conditions resulted in a predicted xylose yield of 9.10g xylose/100g (dry weight) untreated biomass and a glucose yield of 23.34g glucose/100g (dry weight) untreated biomass. The low xylose yields observed were attributed to the high amounts of xylan lost due to solubilization. The quadratic models were found to be reliable for the prediction of sugar yields within the design space. All predicted values were experimentally confirmed with 95% confidence of the predicted values.

Hydrophobic Lurylamine Modified Heteropoly Acid as an Efficient and Recyclable Catalyst for the Hydrolysis Reaction in Aqueous Solution under Microwave

A new modified phosphotungstic acid (HPW) catalyst (HPW-catal) was synthesized b y a simple mixing method under ambient atmosphere and evaluated for the hydrolysis of sucrose and ethyl acetate under microwave irradiation. The as-synthesized catalyst was characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy(FTIR), and NH3-TPD, which proved that HPW was sucessfully modified with lurylamine and the catalyst still remains the Keggin structure. Significantly, the HPW-catal showed excellent catalytic hydrolysis perforance for the sucrose and ethyl acetate under microwave irradiation. Both of the reactions were studied over other solid acid catalysts, such as AM-15 and ZSM-5. At mild tempretures, the sucrose and ethyl acetate could be hydrolyzed into corresponding products. The maximum glucose yield and alcohol could reach 100% and 95%, respectively. The HPW-catal can be recycled and reused. The HPW-catal will provides new ideas for the modifying of heteropoly acids.

CO-PRODUCTION OF LACTIC ACID AND ETHANOL USING RHIZOPUS SP. FROM HYDROLYZED INEDIBLE CASSAVA STARCH AND LEAVES

Production of lactic acid and ethanol from inedible cassava starch and leaves was investigated. Prior to fermentation, hydrolysis of the starch and leaves was conducted. Hydrolysis was optimized by manipulating at three levels each four particular factors, namely acid concentration, starch concentration, temperature, and reaction time. Maximum glucose yield of 0.96 g/g was obtained when 2.5% (w/v) of cassava leaves with 2.5% (w/v) of starch was hydrolyzed using 0.20 M of nitric acid at 160oC of temperature for 10 min. The potential of hydrolyzed cassava starch and leaves was then investigated for lactic acid production using fungal Rhizopus sp. The fermentation process was then conducted in shake flask by varying four factors at three levels each. Maximum lactic acid and ethanol yields of 0.95 g/g and 0.52 g/g, respectively, were achieved at different optimum conditions.  Lactic acid production was found to be linked to a decrease in ethanol production. Design Expert v6.0.8 was used to aid in the design of the experiment using Taguchi’s methodology.

Optimization of hydrothermal pretreatment for co-utilization C-5 and C-6 sugars of cassava alcohol residue

Hydrothermal reaction was first applied to pretreat cassava alcohol residue for realizing the co-utilization of xylose and glucose to assume fermentation ethanol. This work focused on the influence of hydrothermal pretreatment conditions on ethanol production. Hydrothermal reaction was used to explore the maximum xylose and glucose yields, in respect to reaction temperature (120–240°C), solid-liquid ratio (0.023–0.150) and reaction time (15–120min). The results showed that the suitable conditions were at 180–200°C, for 45–60min and with 10–12.5% solids. In this range, the conjunct of xylose and glucose would reach the maximum, which can make full use of hemicellulose and cellulose in cassava alcohol residue. According to the results, respond surface methodology (RSM) based on Box-Behnken design was used to further optimize the three independent variables for the highest ethanol by co-utilization of xylose and glucose. RSM revealed that the effect of temperature on ethanol production was much more significant than the effect of reaction time and solid-liquid ratio, and the highest ethanol production was 70.6mg/g that was close to the experiment value of 69.5mg/g at 193°C for 51min and with 11.5% solids. Furthermore, the crystallinity and morphology of the untreated and pretreated cassava alcohol residue were investigated to assess the effect of hydrothermal pretreatment by scanning electron microscope (SEM) and X-ray diffraction (XRD), respectively.

Comparison of hydrothermal, hydrotropic and organosolv pretreatment for improving the enzymatic digestibility of bamboo

Pretreatment is the crucial step to disrupt the recalcitrant structure of lignocellulosic biomass for improving the enzymatic hydrolysis efficiency. Typically, hydrothermal, organosolv and hydrotropic pretreatments are environmentally benign and effective methods. In this work, effects of hydrothermal, organosolv and hydrotropic pretreatments on improving enzymatic hydrolysis of bamboo were comprehensively compared. Hydrotropic pretreatment was more effective in removal lignin and xylose from bamboo fiber cell wall. However, the surface coverage by lignin and extractives were dramatically displaced during organosolv pretreatment as investigation by X-ray photoelectron spectroscopy. After pretreatments, the crystallinity of cellulose in pretreated substrates has a significant reduction, and pores were exposed on fiber surface. The residual content of acetyl and phenolic groups in hydrotropic pretreated substrates is lower than organosolv pretreated substrates. In order to deeply assess the delignification of pretreatments, the isolated lignins obtaining from pretreatments process were characterized by Fourier transform infrared spectroscopy also. It was revealed that hydrotropic lignin contained more phenolic hydroxyl group and syringyl units than organosolv lignin. Compared to hydrothermal and organosolv pretreatment, cellulase adsorption capacity of pretreated substrates was notably improved by hydrotropic pretreatment, which indicating the better enzyme accessibility of cellulose. Eventually, the maximum glucose yield was obtained from hydrotropic pretreated substrates.

Kraft GL-ethanol pretreatment on sugarcane bagasse for effective enzymatic hydrolysis

Kraft green liquor (GL) mainly consisting of sodium carbonate and sodium sulfide are available from Kraft mills. Kraft GL combined with ethanol (Kraft GL-ethanol) was chosen to pretreat sugarcane bagasse for effective enzymatic hydrolysis. And the obtained lignins and cellulose rich fractions were characterized in detail. The highest lignin removal of 95.3% was observed at a pretreatment temperature of 160°C, and approximately 70.8% of lignin was removed at the pretreatment temperature of 100°C. The lignin removal with Kraft GL-ethanol pretreatment was higher than that with Soda GL-ethanol pretreatment at 140°C. An FT-IR study of the isolated lignins showed that an increase in pretreatment temperature resulted in the cleavage of ester bonds. GPC results revealed that the molecular weight decreased with increasing pretreatment temperature. The 2D HSQC NMR data showed that the cleavage of β-O-4 became more severe with increasing pretreatment temperature from 80 to 160°C. Fewer condensation reactions between lignin units occurred when the pretreatment temperature was above 100°C. Moreover, the enzymatic hydrolysis efficiency of sugarcane bagasse increasing with increasing pretreatment temperature. The glucose yield reached 65.13% at a pretreatment temperature of 80°C. The maximum glucose yield (98.26%) of sugarcane bagasse after 72h of enzymatic hydrolysis was achieved at a pretreatment temperature of 160°C. When lignin content was low in samples after pretreatment at 140–160°C, the enzymatic hydrolysis efficiency of the substrate increased because the number of crystalline cellulose regions decreased. Scanning electron microscopy showed that the surfaces of Kraft GL-ethanol pretreated sugarcane bagasse were separated into individual fibers. In addition, contact angle analysis showed that the Kraft GL-ethanol pretreated sugarcane bagasse was more hydrophilic than untreated sugarcane bagasse.

Waste Bread as a Biomass Source: Optimization of Enzymatic Hydrolysis and Relation between Rheological Behavior and Glucose Yield

Amongst the many forms of food waste, bread is a major contributor to the problem. Two aims of this research investigate waste breads potential of being a bioresource for the production of fermentable sugars which are precursor of valuable bioproducts by fermentation process: finding optimum substrate, water and enzyme ratio to produce the highest amount of fermentable sugars and investigating the rheological behavior of the system during hydrolysis. Two stage waste bread hydrolysis was performed with enzymes α-amylase and amyloglucosiades and response surface methodology was used to optimize substrate, water and enzyme ratio. Discovery Hybrid Rheometer-2 (TA Instruments) fitted with a parallel-plate geometry was used to investigate steady flow viscosity of the slurry during hydrolysis. 99 % of theoretical maximum glucose yield, a main fermentable sugar, is achieved by optimizing enzymatic hydrolysis conditions of waste bread at liquefaction and saccharification stages. Just after the addition of α-amylase enzyme, substantial decrease is observed in viscosity and Casson apparent yield stress of the slurry. During saccharification stage, glucose yield increases dramatically while viscosity of the slurries is very low and does not change considerably. The results imply that utilizing high concentrations of waste bread as a feedstock to produce fermentation products offers economic benefits without causing high power consumption, excessive wear on equipment, and reduced conversion which are generally expected consequences for high-solid processing.

A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis

The enzymatic hydrolysis of cellulose and lignocellulosic materials is marked by a rate decrease along the reaction time. Cellobiohydrolase slow dissociation from the substrate and its inhibition by the cellobiose produced are relevant factors associated to the rate decrease. In that sense, addition of β-glucosidases to the enzyme cocktails employed in cellulose enzymatic hydrolysis not only produces glucose as final product but also reduces the cellobiohydrolase inhibition by cellobiose. The digestive β-glucosidase GH1 from the fall armyworm Spodoptera frugiperda, hereafter called Sfβgly, containing the mutation L428V showed an increased kcat for cellobiose hydrolysis. In comparison to assays conducted with the wild-type Sfβgly and cellobiohydrolase TrCel7A, the presence of the mutant L428V increased in 5 fold the initial rate of crystalline cellulose hydrolysis and reduced to one quarter the time needed to TrCel7A produce the maximum glucose yield. As our results show that mutant L428V complement the action of TrCel7A, the introduction of the equivalent replacement in β-glucosidases is a promising strategy to reduce costs in the enzymatic hydrolysis of lignocellulosic materials.

A biotechnological process efficiently co-produces two high value-added products, glucose and xylooligosaccharides, from sugarcane bagasse

In this study, a co-production of two high value-added products, glucose and xylooligosaccharides (XOS), was investigated by utilizing sugarcane bagasse (SB) within a multi-product bio-refinery framework optimized by Box–Behnken design-based response surface methodology. The developed process resulted in a maximum cellulose conversion of xylan-removed SB, 98.69±1.30%, and a maximum extracted SB xylan conversion into XOS (xylobiose and xylotriose) of 57.36±0.79% that was the highest SB xylan conversion reported in the literature, employing cellulase from Penicillium oxalicum EU2106 and recombinant endo-β-1,4-xylanase in Pichia pastoris. Consequently, a mass balance analysis showed that the maximum yields of glucose and XOS were 34.43±0.32g and 5.96±0.09g per 100g raw SB. Overall, this described process may be a preferred option for the comprehensive utilization of SB.

Alkaline-sulfite pretreatment and use of surfactants during enzymatic hydrolysis to enhance ethanol production from sugarcane bagasse.

Sugarcane bagasse is a by-product from the sugar and ethanol industry which contains approximately 70 % of its dry mass composed by polysaccharides. To convert these polysaccharides into fuel ethanol it is necessary a pretreatment step to increase the enzymatic digestibility of the recalcitrant raw material. In this work, sugarcane bagasse was pretreated by an alkaline-sulfite chemithermomechanical process for increasing its enzymatic digestibility. Na2SO3 and NaOH ratios were fixed at 2:1, and three increasing chemical loads, varying from 4 to 8 % m/m Na2SO3, were used to prepare the pretreated materials. The increase in the alkaline-sulfite load decreased the lignin content in the pretreated material up to 35.5 % at the highest chemical load. The pretreated samples presented enhanced glucose yields during enzymatic hydrolysis as a function of the pretreatment severity. The maximum glucose yield (64 %) was observed for the samples pretreated with the highest chemical load. The use of 2.5 g l(-1) Tween 20 in the hydrolysis step further increased the glucose yield to 75 %. Semi-simultaneous hydrolysis and fermentation of the pretreated materials indicated that the ethanol yield was also enhanced as a function of the pretreatment severity. The maximum ethanol yield was 56 ± 2 % for the sample pretreated with the highest chemical load. For the sample pretreated with the lowest chemical load (2 % m/m NaOH and 4 % m/m Na2SO3), adding Tween 20 during the hydrolysis process increased the ethanol yield from 25 ± 3 to 39.5 ± 1 %.

Fungal pretreatment of non-sterile miscanthus for enhanced enzymatic hydrolysis

Miscanthus was pretreated with the fungus Ceriporiopsis subvermispora under non-sterile conditions, using sterile miscanthus that had been previously colonized with the fungus as the inoculum. Inoculum ratios equal to or greater than 30% yielded a successful pretreatment, enhancing the enzymatic digestibility of miscanthus by 3- to 4-fold over that of raw miscanthus, which was comparable with the fungal pretreatment under sterile conditions. This enhanced digestibility was linearly correlated with lignin degradation. Although cellulose loss of up to 13% was observed for the successful non-sterile pretreatments, the final glucose yield was 3–4 times higher than that of raw miscanthus and comparable to that of the sterile pretreated miscanthus. A time course study showed that maximum glucose yield can be achieved with a pretreatment time of 21days.

Effects of Pre-Treatment on Lignocellulosic Butanol as a Bio-Fuel Produced from Bamboo Using <i>Clostridium acetobutylicum</i>

Conversion of lignocellulosic biomass from bamboo (Bambusa vulgaris) to butanol is an important alternative energy source. In this work, bamboo was used as biomass feedstock for the production of butanol by the fermentation of sugars. Mechanical grinding was carried out, followed by pre-treatment with dilute sulfuric acid concentration of 0.5 and 1.0 (%v/v). This was done at temperatures of 25, 110, 120, 150 and 200°C at time intervals of 2 and 4 hours. Pre-hydrolysate was later analyzed for total sugars by the use of UV-Visible Spectrophotometer. For the conditions considered, the maximum glucose yields were obtained at 200°C. The yields after pre-treatment were 244.80 mg/g, at pre-treatment conditions of 200°C and acid concentrations of 1% for 4 hours. Water insoluble solids obtained were subsequently hydrolysed with Celluclast (Trichoderma reesi) and β-glucosidase (Novozyme 188) for 72 hrs. Bacteria (Clostridium acetobutylicum) were then used to ferment the solubilized sugar into butanol. Raman spectroscopy was used to determine the butanol yield. Optical Microscope images of bamboo samples were obtained at various stages of pre-treatment and enzymatic hydrolysis. These revealed the morphological changes that occur in the cellular structure of the bamboo during exposure to acid and enzymatic hydrolysis. The results show that, increasing temperature, time and acid concentration are associated with higher total sugar yields and cellulose conversion rates. 10.4mg/mL of butanol was produced in sample treated at 1% H2SO4 for 110°C.