November 2018 Arup Jyoti Borah (Roll No Center for Energy Indian Institute of Technology Guwahati Guwahati, Assam, India. Arun Goyal Professor (MTech, PhD) (FAMI, FABAP, FBRS, FNABS,FNAAS,FIFIB) Afdeling Biowetenschappen en Bio-engineering Indian Institute van technologie Guwahati Guwahati Assam, India.
Physical Insights of Ultrasound-Assisted Ethanol Production from Composite feedstock of Invasive
157 5.4.2 Experimental results of ethanol fermentation 157 5.4.3 Simulation results of ethanol fermentation 158 5.5 Flow cytometry analysis of cells used in fermentations 162.
Mechanistic Investigations in Biobutanol Synthesis via Ultrasound–Assisted ABE fermentation using
Overview and Suggestions for Future Work 199
Results of mixture design with biomass obtained from Table 4.2 (A). B) Results of mixture design with biomass obtained from table 4.2. Figure 6.2 Time profiles of the pH of the fermentation broth under control. mechanical shaking) and test (mechanical shaking + ultrasonication.
I NTRODUCTION AND L ITERATURE R EVIEW
Introduction
India’s approach to biofuel policy
The policy also envisages the development of the next generation of more efficient technologies for the conversion of biofuels based on new raw materials. The policy will bring accelerated development and promotion of cultivation, production and use of biofuels, which will increasingly replace gasoline and diesel in transport.
Invasive weedy lignocellulosic biomass
- Global and national scenario
Globally, losses due to IAS invasion have been estimated at US$1.4 trillion per year (Table 1.2), which is close to 5% of global GDP (gross domestic product) (Avery 2013). A detailed account of weed invasion in the main grassland and national park of Assam is shown in Table 1.3 and Table 1.4 below.
Pretreatment and hydrolysis of biomasses
- Physical pretreatment .1 Mechanical comminution
- Physicochemical pretreatment .1 Steam explosion
- Ammonia fiber explosion (AFEX)
- Carbon dioxide explosion
- Chemical pretreatment .1 Ozonolysis
- Acid pretreatment
- Alkaline pretreatment
- Oxidative delignification
- Organosolv process
- Biological pretreatment
This provides access for acids and enzymes to hydrolyze cellulose by increasing the porosity of the lignocellulosic material so that the cellulose is easily accessible and hydrolyzed. This process is used to reduce the size of materials, usually 10-30mm after shredding, to 0.2-2mm by milling or grinding.
Dilute acid hydrolysis
Detoxification of lignocellulose hydrolysate after acid hydrolysis
- Phenolic compounds
- Aliphatic acids
- Furan aldehydes
- Inorganic compounds
Some aromatic carboxylic acids can act as sequestrants, as shown by experiments with plant cells and salicylic acid (Norman et al., 2004). In contrast, another aromatic carboxylic acid, p-hydroxybenzoic acid, which is common in lignocellulosic hydrolysates, does not show a separation effect (Norman et al., 2004).
Delignification of acid hydrolyzed biomasses
- Ultrasound induced delignification of acid hydrolyzed biomass
Simultaneous analysis of experimental and simulation results reveals interesting mechanistic facets of the delignification process, as detailed in the subsequent sections. A summary of some representative studies on the optimization of pretreatment and fermentation of the invasive weeds is given in Table 1.5.
Saccharification of delignified biomass
Most of the studies are aimed at reducing the recalcitrance of cellulose and making it susceptible to enzymatic hydrolysis and facilitating the formation of a fermentable form of sugar by reducing crystallinity and increasing the accessibility to the glucan chain. Both sonication and cavitation create intense energy concentration on a temporal and spatial scale, which improves the kinetics of the process.
Fermentation of enzymatic hydrolysate to ethanol
The main limitation regarding SSF is the compromise with optimal conditions of enzymatic hydrolysis and fermentation. Evaluation of the effects of ultrasound on the enzymatic hydrolysis of sugarcane bagasse to obtain fermentable sugars.
Butanol as a potential biofuel
- Butanol production from lignocellulosic biomass
- ABE Fermentation: past and current perspectives
- ABE fermentation process
- Microbes role in ABE fermentation
- Commercialization of butanol production
- Process design in ABE fermentation
- Sensitivity analysis of butanol production through ABE fermentation
The low butanol yield and butanol concentration with low cell densities made butanol production from glucose by ABE fermentation uneconomic (Maddox, 1989). Various challenges related to ABE fermentation and their solution that will lead to commercialization are shown in Table 1.10.
Comparison of process economics of ethanol and butanol
Acceleration of the bioprocess by ultrasound treatment
Biorefinery approach
Scope of the present study and specific objectives
Techno-Economic Analysis of the Production of Cellulosic Butanol from corn starch through acetone-butanol-ethanol fermentation. Direct ethanol production from lignocellulosic sugars and sugarcane bagasse by a recombinant strain Trichoderma reesei HJ48. Butanol production from wheat straw by simultaneous saccharification and fermentation using Clostridium beijerinckii: Part I - Batch fermentation.
WEEDS AS MULTIPLE FEEDSTOCKS FOR BIOFUEL PRODUCTION
AN ASSESSMENT OF THE POTENTIAL OF INVASIVE WEEDS AS MULTIPLE FEEDSTOCKS FOR BIOFUEL
Introduction
- Aim and approach
In the present study, we performed pretreatment of the eight invasive weeds mentioned above at optimized conditions determined for Parthenium hysterophorus, (Singh et al., 2014) and assessed the yield of reducible sugars. In such a situation, it may not be possible or practical to carry out a comprehensive optimization of the pretreatment conditions for each biomass used as feedstock. The present study essentially attempts to paint a picture of such variations by pretreatment of the eight selected invasive weeds under conditions optimized for the weed of Parthenium hysterophorus.
Materials and methods
- Biomass collection and processing
- Proximate and biochemical analysis of invasive biomasses
- Acid hydrolysis of invasive biomasses
- Detoxification of acid hydrolysate of invasive biomass
- Delignification of acid pretreated invasive biomasses
- Saccharification of invasive biomasses by commercial enzymes
- Determination of reducing sugar in acid and enzyme hydrolysate
- Characterization of the raw, acid pretreated and delignified biomasses
After completion of the pretreatment, the biomass was separated from the reaction mixture by filtration through a double-layered muslin cloth. This was followed by drying the biomass residue in a hot air oven for 24 hours at 60 °C. The delignification of the biomass obtained after acid hydrolysis was carried out using the procedure outlined by (Bharadwaja et al., 2015).
Results and discussion
- Results of dilute acid and enzymatic hydrolysis
- Enzymatic hydrolysis
- Total fermentable sugar (TFS) yield
- Assessment of biofuels production potential
- Results of biomass characterization
The results of enzymatic hydrolysis of the pretreated biomasses of the eight invasive weed species are depicted in fig. An increase in crystallinity index essentially indicates an increase in the percentage of cellulose content in biomass after pretreatment. However, the extent of the increase in crystallinity is different for the eight biomass species.
Conclusion
Mechanistic investigation into ultrasound-induced enhancement of enzymatic hydrolysis of invasive biomass species, Bioresource Technology. Effects of pretreatment on the morphology, chemical composition and enzymatic digestibility of eucalyptus bark: a potentially valuable source of fermentable sugars for biofuel production – Part 1. A correction to 'Feasibility of orice straw as alternative substrate for biobutanol production' by Ranjan et al.
INVASIVE BIOMASS SPECIES §
MECHANISTIC INVESTIGATION IN ULTRASOUND INDUCED ENHANCEMENT OF ENZYMATIC
Introduction
A peculiarity of the present study is that enzymatic treatments of all biomasses were carried out under constant conditions. The pretreatment of the invasive weeds prior to enzymatic hydrolysis was also performed under constant conditions, which were optimized for P. The rationale underlying this strategy is explained next. Previous authors have reported studies in ultrasonic enhancement of the enzymatic hydrolysis of pretreated biomass.
Materials and methods .1 Materials
- Pretreatment of raw biomass and hydrolysis of pretreated biomass
- Protocol of enzymatic hydrolysis of biomass with mechanical shaking
- Protocol for enzymatic hydrolysis of biomass with sonication
- Total reducing sugar estimation
- Analysis of enzyme structure
The progress of the enzymatic hydrolysis was monitored by periodically withdrawing 0.1 mL samples of the reaction mixture and analyzing for sugar release. The temperature of the reaction mixture inside the flask was the same as that of the water outside. Similar to control experiments, the progress of sugar release in enzymatic hydrolysis was monitored by periodically withdrawing 0.1 mL portions of the reaction mixture and analyzing them for reducing sugar concentration.
Mathematical model of enzyme hydrolysis of delignified biomass
The numerical values of these parameters are obtained by comparing the numerical solution of the ordinary differential equation (ODE) for reaction rate with the experimental profile of total reducing sugar (ie, the soluble product, Gs). For calculating the simulated profile of soluble hydrolysis product and matching it with experimental profile, optimal values of the model parameters in Eq. The optimal values of the model parameters were determined by calculating the root mean square (RMS) error between experimental values and model results using genetic algorithm (GA) (Flowchart given in Appendix B Fig.
Results and Discussion
- Experimental results of enzymatic hydrolysis
- Simulation results of enzymatic hydrolysis using HCH-1 model
- Results of intrinsic fluorescence and circular dichroism analysis
The results of enzymatic hydrolysis simulations using the HCH-1 model are shown in Table 3.2, where the values of the model parameters under the conditions are listed. Enzymatic hydrolysis simulation results: Kinetic and physiological parameter values in the HCH–1 model. The practically identical values of during mechanical mixing and sonication show that enzyme adsorption on cellulose is not limited by mass transfer.
Analysis of the circular dichroism spectrum using the Dichroweb online server is given in Table 3.3. Reduction of the -helix structure is more pronounced for ultrasound (or sonication) treatment than mechanical agitation. Intrinsic spectra of Cellobiase Figure 3.4 Intrinsic fluorescence spectra of enzymes in different forms (enzyme treatment and post-. with mechanical shaking and sonication at atmospheric pressure or 101.3 kPa).
Conclusion
The enhancing effect of sonication was found to be independent of the conditions of enzymatic hydrolysis – optimized or non-optimized. Mechanistic insight into ultrasound-induced enhancement of the simultaneous saccharification and fermentation of Parthenium hysterophorus for ethanol production. Estimation of protein secondary structure from circular dichroism spectra: comparison of CONTIN, SELCON and CDSSTR methods with an extended reference set.
FEEDSTOCK FOR BIOFUEL PRODUCTION
KINETIC MODELING OF DILUTE ACID HYDROLYSIS OF VARIOUS INVASIVE WEEDS TO DEVELOP A
Introduction
- Assessment of biofuels production potential
One of the best ways to reduce the impact of such an invasive weed on the ecosystem is to exploit its biomass as a raw material for the production of biofuels. The best four of the eight biomasses were considered as Lot 1 for the mixed design, and the remaining four biomasses were placed in Lot 2. The best lot from each set (category) will be mixed to achieve the best ratio of the eight biomass combination in terms of TRS yield.
Materials and Methods
- Materials
- Biomass collection and processing
- Protocol for hemicellulose estimation
- Dilute acid pretreatment of raw biomasses, delignification and enzymatic hydrolysis of composite biomass
- Characterization of raw and pretreated composite biomass
- Kinetic Modeling to determine the rate constant of acid hydrolysis
The crystallinity of the cellulose residue in the composite biomass was calculated according to the empirical method proposed by Segal et al. The solver function works by changing the values of k1 and k2 to try to get an error value of zero. The concentrations of the sugars produced, which are obtained by analyzing the chromatograms, are determined as the initial concentrations of cellulose and hemicellulose.
Results and Discussion
- Kinetic modeling of acid hydrolysis
- Biochemical analysis of composite biomass
- Kinetic assessment of enzymatic hydrolysis
- Results of composite biomass characterization
This assumption is clearly reflected from the k1 value of E. Time required for this bioconversion of TRS is almost reduced to 3.5 times for E. The outcome of the results can be attributed to the biochemical constituent of the lignocellulose. The data obtained from acid and enzymatic hydrolysis of the composite biomass (both category A and B) are given in Table 4.4(A). The results of TRS yield of the delignified composite biomass after enzyme hydrolysis for both mechanical agitation (control) and ultrasound agitation (test) are shown in Fig.
Conclusion
Physical and chemical characteristics of corn stover and poplar solids resulting from leading pretreatment technologies. Butanol Production Using Clostridium beijerinckii BA 101 Hyper–Butanol-Producing Mutant Strain and Recovery by Pervaporation.
INVASIVE WEEDS §
PHYSICAL INSIGHTS OF ULTRASOUND-ASSISTED
Introduction
Ultrasound is an established technique for the intensification of various physical and chemical processes and has also been used for improved bioethanol production (Nikolic et al., 2010; Subhedar et al., 2015; Battista et al., 2016; Berlowska et al. ., 2016; ). Ultrasound exerts its effect on the feedback system through implosive collapse cavitation bubbles that create intense energy concentration on an extremely small spatial and temporal scale (Nalajala et al., 2010; Choudhury et al., 2014). The main physical impact of ultrasound and cavitation is the generation of intense microturbulence that imparts micromixing in the medium, which overcomes mass transfer limitations.
Materials and Methods
- Chemicals and reagents
- Biomass collection and its processing
- Pretreatment of raw composite biomass (acid pretreatment and delignification)
- Enzymatic hydrolysis
- Microorganism, culture revival and maintenance
- Fermentation of composite biomass for ethanol production
- Analysis
- Viability analysis of sonication-exposed yeast cells by FACS
The cellulose content of the delignified solids was determined as g/g of delignified biomass according to standard TAPPI protocols (Allan et al., 1992). The position of the flask was carefully kept similar in all experiments performed to avoid artifacts generated due to fluctuating ultrasound intensity (Moholkar et al., 2000). Rest of the protocol for the control and test experiments was the same as described earlier for hexose fermentation (Singh et. al., 2015).
Mathematical model
Total reducing sugars (TRS): The consumption of TRS in the fermentation broth occurs through two mechanisms viz. The numerical solution of the above equations was compared with the experimental time profiles of concentration, ethanol concentration and cell mass concentration.
Results and Discussion
- Acid hydrolysis and enzymatic hydrolysis of composite biomass
- Experimental results of ethanol fermentation
- Simulation results of ethanol fermentation
The values of the kinetic and physiological parameters of the fermentation model obtained after fitting the simulated profiles to the experimental data by applying GA optimization are given in Table 5.2. Due to the reduced energy requirements for cell maintenance, a greater proportion of the substrate is used for cell metabolism resulting in increased ethanol productivity. The additional beneficial effect of microconvection is the deagglomeration of the cells and the regulation of the osmolarity of the internal volume of the cell.
Flow cytometry analysis of cells used in fermentations
Of the contour plots shown in (Fig. 5.3), the upper left quadrant (Q1) indicates the living population, and the upper right quadrant (Q2) consists of membrane compromised cells. Dead cells appeared in the lower right quadrant (Q3) and unstained cells in the lower left quadrant (Q4). These contour plots are machine generated based on the relative intensity collected in the two filter chambers, viz.
Conclusion
Multiparametric flow cytometry and cell sorting for the assessment of viable, injured and dead Bifidobacterium cells during bile salt stress. Optimization of process variables using response surface methodology (RSM) for ethanol production from cashew apple juice by Saccharomyces cerevisiae. Mathematical modeling of cellulose conversion to ethanol by simultaneous saccharification and fermentation process, Applied Biochemistry and Biotechnology.
USING MIXED FEEDSTOCK OF INVASIVE WEEDS §
MECHANISTIC INVESTIGATIONS IN BIOBUTANOL SYNTHESIS VIA ULTRASOUND–ASSISTED ABE
Introduction
As a solution to the high cost of the substrate (Demirbas, 2009, Cherubini et al. 2010, Farmanbordara et al. 2018), the present study has used mixed (or composite) biomass of several invasive weeds as feedstock for fermentation . However, these weeds are high in holocellulose (in the form of hemicellulose and cellulose), which can be hydrolyzed to produce fermentable pentose and hexose sugars (Singh et al. Votruba et al., (1986) synthesized the equations of model on kinetic rate analysis of batch fermentation data under optimal conditions of media composition and physical parameters.
Materials, methods and mathematical model
- Chemicals and reagents
- Composite biomass
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