Application of Thermoplastic Microbioreactor to the Single Cell Study of Budding Yeast to Decipher the Effect of 5-Hydroxymethylfurfural on Growth

Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Gasification of Trans-4-Hydroxycinnamic Acid with Ethanol at Elevated Temperatures

Lignin is a major constituent of woody biomass, and exists abundantly in nature. It is the major byproducts from the paper industry and bioethanol production processes. The byproducts are mainly used for low-valued applications. Instead, lignin can be converted into higher-valued gaseous fuel, thereby helping to curtail the ever-growing price of oil and to slow down the trend of global warming. Although biochemical treatment is capable of converting cellulose into liquid ethanol fuel, it cannot be applied to the conversion of lignin. Alternatively, it is possible to convert lignin into gaseous fuel thermochemically. In the present work, trans-4-hydroxycinnamic acid, a model compound for lignin, which closely resembles the basic building blocks of lignin, is gasified in an autoclave with ethanol at elevated temperatures and pressures, that are above the critical point of ethanol. Ethanol, instead of water, is chosen, because ethanol dissolves trans-4-hydroxycinnamic acid easily and helps to convert it into lighter gaseous species relatively well. The major operating parameters for the gasification reaction include temperature (673-873 K), reaction pressure (5-25 MPa) and feed concentration (0.05-0.3 M). Generally, more than 80% of the reactant, including trans-4-hydroxycinnamic acid and ethanol, were converted into gaseous products at an operating condition of 873 K and 5 MPa.

Processing and Economic Analysis of Rain Tree (Samanea saman) Pods for Village Level Hydrous Bioethanol Production

Biofuel is one of the renewable energy sources adapted by the Philippine government in order to lessen the dependency on foreign fuel and to reduce carbon dioxide emissions. Rain tree pods were seen to be a promising source of bioethanol since it contains significant amount of fermentable sugars. The study was conducted to establish the complete procedure in processing rain tree pods for village level hydrous bioethanol production. Production processes were done for village level hydrous bioethanol production from collection, drying, storage, shredding, dilution, extraction, fermentation, and distillation. The feedstock was sundried, and moisture content was determined at a range of 20% to 26% prior to storage. Dilution ratio was 1:1.25 (1 kg of pods = 1.25 L of water) and after extraction process yielded a sugar concentration of 22 0Bx to 24 0Bx. The dilution period was three hours. After three hours of diluting the samples, the juice was extracted using extractor with a capacity of 64.10 L/hour. 150 L of rain tree pods juice was extracted and subjected to fermentation process using a village level anaerobic bioreactor. Fermentation with yeast (Saccharomyces cerevisiae) can fasten up the process, thus producing more ethanol at a shorter period of time; however, without yeast fermentation, it also produces ethanol at lower volume with slower fermentation process. Distillation of 150 L of fermented broth was done for six hours at 85 °C to 95 °C temperature (feedstock) and 74 °C to 95 °C temperature of the column head (vapor state of ethanol). The highest volume of ethanol recovered was established at with yeast fermentation at five-day duration with a value of 14.89 L and lowest actual ethanol content was found at without yeast fermentation at three-day duration having a value of 11.63 L. In general, the results suggested that rain tree pods had a very good potential as feedstock for bioethanol production. Fermentation of rain tree pods juice can be done with yeast and without yeast.

Kinetic Parameters for Bioethanol Production from Oil Palm Trunk Juice

Abundant and cheap agricultural waste of oil palm trunk (OPT) juice was used to produce bioethanol. Two strains of Saccharomyces cerevisiae and a strain of Pichia stipitis were used to produce bioethanol from the OPT juice. Fermentation was conducted at previously optimized condition at 30oC and without shaking. The kinetic parameters were estimated and calculated. Monod equation and Hinshelwood model is used to relate the specific growth to the concentration of the limiting substrate and also to simulate bioethanol production rate. Among the three strains, single S. cerevisiae Kyokai no. 7 produce the highest ethanol yield of 0.477 g/l.h within the shortest time (12 h). This yeast also produces more than 20 g/l ethanol concentration within 10 h of fermentation.

Fermentable Sugars from Palm Empty Fruit Bunch Biomass for Bioethanol Production

This study investigated the effect of a dilute acid, lime and ammonia aqueous pretreatment on the fermentable sugars conversion from empty fruit bunch (EFB) biomass. The dilute acid treatment was carried out in an autoclave, at 121ºC with 4% of sulfuric acid. In the lime pretreatment, 3 wt % of calcium hydroxide was used, whereas the third method was done by soaking EFB with 28% ammonia solution. The EFB biomass was then subjected to a two-stage-acid hydrolysis process. Subsequently, the hydrolysate was fermented by using instant baker’s yeast to produce bioethanol. The highest glucose yield was 890 mg/g of biomass, obtained from the sample which underwent lime pretreatment. The highest bioethanol yield of 6.1mg/g of glucose was achieved from acid pretreatment. This showed that the acid pretreatment gave the most fermentable sugars compared to the other two pretreatments.

Thermal and Morphological Evaluation of Chemically Pretreated Sugarcane Bagasse

Enzymatic hydrolysis is one of the major steps involved in the conversion from sugarcane bagasse to yield ethanol. This process offers potential for yields and selectivity higher, lower energy costs and milder operating conditions than chemical processes. However, the presence of some factors such as lignin content, crystallinity degree of the cellulose, and particle sizes, limits the digestibility of the cellulose present in the lignocellulosic biomasses. Pretreatment aims to improve the access of the enzyme to the substrate. In this study sugarcane bagasse was submitted chemical pretreatment that consisted of two consecutive steps, the first with dilute sulfuric acid (1 % (v/v) H2SO4), and the second with alkaline solutions with different concentrations of NaOH (1, 2, 3 and 4 % (w/v)). Thermal Analysis (TG/ DTG and DTA) was used to evaluate hemicellulose, cellulose and lignin contents in the samples. Scanning Electron Microscopy (SEM) was used to evaluate the morphological structures of the in natura and chemically treated samples. Results showed that pretreatments were effective in chemical degradation of lignocellulosic materials of the samples, and also was possible to observe the morphological changes occurring in the biomasses after pretreatments.

Biodegradation of Lignocellulosic Residues of Water Hyacinth (Eichhornia crassipes) and Response Surface Methodological Approach to Optimize Bioethanol Production Using Fermenting Yeast Pachysolen tannophilus NRRL Y-2460

The objective of this research was to investigate biodegradation of water hyacinth (Eichhornia crassipes) to produce bioethanol using dilute-acid pretreatment (1% sulfuric acid) results in high hemicellulose decomposition and using yeast (Pachysolen tannophilus) as bioethanol producing strain. A maximum ethanol yield of 1.14g/L with coefficient, 0.24g g-1; productivity, 0.015g l-1h-1 was comparable to predicted value 32.05g/L obtained by Central Composite Design (CCD). Maximum ethanol yield coefficient was comparable to those obtained through enzymatic saccharification and fermentation of acid hydrolysate using fully equipped fermentor. Although maximum ethanol concentration was low in lab scale, the improvement of lignocellulosic ethanol yield is necessary for large scale production.

Investigation of Pre-Treatment Parameters of Rye and Triticale for Bioethanol Production

This paper presents the new results of energy plant – rye and triticale at yellow ripeness and ripe, pre-treatment in high pressure steam reactor and monosaccharide extraction. There were investigated the influence of steam pressure (20 to 22 bar), retention duration (180 to 240 s) and catalytic sulphuric acid concentration strength (0 to 0.5 %) on the pre-treatment process, contents of monosaccharides (glucose, arabinose, xylose, mannose) and undesirable by-compounds (furfural and HMF) in the reactor. The study has determined that the largest amount of monosaccharides (37.2 % of glucose, 2.7 % of arabinose, 8.4 % of xylose, and 1.3 % of mannose) was received in the rye at ripe, the samples of which were mixed with 0.5 % concentration of catalytic sulphuric acid, and hydrolysed in the reactor, where the pressure was 20 bar, whereas the reaction time – 240 s.

Bioethanol - A Viable Answer to India-s Surging Energy Needs

India is currently the second most populous nation in the world with over 1.2 billion people, growing annually at the rate of 1.5%. It is experiencing a surge in energy demands, expected to grow more than three to four times in 25 years. Most of the energy requirements are currently satisfied by the import of fossil fuels – coal, petroleum-based products and natural gas. Biofuels can satisfy these energy needs in an environmentally benign and cost effective manner while reducing dependence on import of fossil fuels, thus providing National Energy Security. Among various forms of bioenergy, bioethanol is one of the major options for India because of availability of feed stock crops. This paper presents an overview on bioethanol production and technology, steps taken by the Indian government to facilitate and bring about optimal development and utilization of indigenous biomass feedstocks for production of this biofuel.

The Kinetic of Biodegradation Lignin in Water Hyacinth (Eichhornia Crassipes) by Phanerochaete Chrysosporium using Solid State Fermentation (SSF) Method for Bioethanol Production, Indonesia

Lignocellulosic materials are considered the most abundant renewable resource available for the Bioethanol Production. Water Hyacinth is one of potential raw material of the world-s worst aquatic plant as a feedstock to produce Bioethanol. The purposed this research is obtain reduced of matter for biodegradation lignin in Biological pretreatment with White Rot Fungi eg. Phanerochaete Chrysosporium using Solid state Fermentation methods. Phanerochaete Chrysosporium is known to have the best ability to degraded lignin, but simultaneously it can also degraded cellulose and hemicelulose. During 8 weeks incubation, water hyacinth occurred loss of weight reached 34,67%, while loss of lignin reached 67,21%, loss of cellulose reached 11,01% and loss of hemicellulose reached 36,56%. The kinetic of losses lignin using regression linear plot, the results is obtained constant rate (k) of reduction lignin is -0.1053 and the equation of reduction of lignin is y = wo - 0, 1.53 x

Sweetpotato Organic Cultivation with Wood Vinegar, Entomopathogenic Nematode and Fermented Organic Substance from Plants

The effect of wood vinegar, entomopathogenic nematodes ((Steinernema thailandensis n. sp.) and fermented organic substances from four plants such as: Derris elliptica Roxb, Stemona tuberosa Lour, Tinospora crispa Mier and Azadirachta indica J. were tested on the five varieties of sweetpotato with potential for bioethanol production ie. Taiwan, China, PROC No.65-16, Phichit 166-5, and Phichit 129-6. The experimental plots were located at Faculty of Agriculture, Natural Resources and Environment, Naresuan University, Phitsanulok, Thailand. The aim of this study was to compare the efficiency of the five treatments for growth, yield and insect infestation on the five varieties of sweetpotato. Treatment with entomopathogenic nematodes gave the highest average weight of sweetpotato tubers (1.3 kg/tuber), followed by wood vinegar, fermented organic substances and mixed treatment with yields of 0.88, 0.46 and 0.43 kg/tuber, respectively. Also the entomopathogenic nematode treatment gave significantly higher average width and length of sweet potato (9.82 cm and 9.45 cm, respectively). Additionally, the entomopathogenic nematode provided the best control of insect infestation on sweetpotato leaves and tubers. Comparison among the varieties of sweetpotato, PROC NO.65-16 showed the highest weight and length. However, Phichit 129-6 gave significantly higher weight of 0.94 kg/tuber. Lastly, the lowest sweet potato weevil infestation on leaves and tubers occurred on Taiwan and Phichit 129-6.

Bioethanol Production from Enzymatically Saccharified Sunflower Stalks Using Steam Explosion as Pretreatment

Sunflower stalks were analysed for chemical compositions: pentosan 15.84%, holocellulose 70.69%, alphacellulose 45.74%, glucose 27.10% and xylose 7.69% based on dry weight of 100-g raw material. The most optimum condition for steam explosion pretreatment was as follows. Sunflower stalks were cut into small pieces and soaked in 0.02 M H2SO4 for overnight. After that, they were steam exploded at 207 C and 21 kg/cm2 for 3 minutes to fractionate cellulose, hemicellulose and lignin. The resulting hydrolysate, containing hemicellulose, and cellulose pulp contained xylose sugar at 2.53% and 7.00%, respectively.The pulp was further subjected to enzymatic saccharification at 50 C, pH 4.8 citrate buffer) with pulp/buffer 6% (w/w)and Celluclast 1.5L/pulp 2.67% (w/w) to obtain single glucose with maximum yield 11.97%. After fixed-bed fermentation under optimum condition using conventional yeast mixtures to produce bioethanol, it indicated maximum ethanol yield of 0.028 g/100 g sunflower stalk.