Abstract: 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.
Abstract: A systematic study was conducted to explore the photocatalytic reduction of carbon dioxide (CO2) into methanol on TiO2 loaded copper ferrite (CuFe2O4) photocatalyst under visible light irradiation. The phases and crystallite size of the photocatalysts were characterized by X-ray diffraction (XRD) and it indicates CuFe2O4 as tetragonal phase incorporation with anatase TiO2 in CuFe2O4/TiO2 hetero-structure. The XRD results confirmed the formation of spinel type tetragonal CuFe2O4 phases along with predominantly anatase phase of TiO2 in the CuFe2O4/TiO2 hetero-structure. UV-Vis absorption spectrum suggested the formation of the hetero-junction with relatively lower band gap than that of TiO2. Photoluminescence (PL) technique was used to study the electron–hole (e−/h+) recombination process. PL spectra analysis confirmed the slow-down of the recombination of electron–hole (e−/h+) pairs in the CuFe2O4/TiO2 hetero-structure. The photocatalytic performance of CuFe2O4/TiO2 was evaluated based on the methanol yield with varying amount of TiO2 over CuFe2O4 (0.5:1, 1:1, and 2:1) and changing light intensity. The mechanism of the photocatalysis was proposed based on the fact that the predominant species of CO2 in aqueous phase were dissolved CO2 and HCO3- at pH ~5.9. It was evident that the CuFe2O4 could harvest the electrons under visible light irradiation, which could further be injected to the conduction band of TiO2 to increase the life time of the electron and facilitating the reactions of CO2 to methanol. The developed catalyst showed good recycle ability up to four cycles where the loss of activity was ~25%. Methanol was observed as the main product over CuFe2O4, but loading with TiO2 remarkably increased the methanol yield. Methanol yield over CuFe2O4/TiO2 was found to be about three times higher (651 μmol/gcat L) than that of CuFe2O4 photocatalyst. This occurs because the energy of the band excited electrons lies above the redox potentials of the reaction products CO2/CH3OH.
Abstract: The effect of Zn2+, Mg2+, and Ba2+ on Saccharomyces
pastorianus performance was evaluated in this study at independent
and three variable combinations. After 96 h of fermentation, high
wort fermentability (%F) = 29.53 was obtained in medium containing
900:4 ppm Mg2+ + Ba2+. Increased ethanol yield 7.35 %(v/v) and
7.13 %(v/v) were obtained in media containing 900:4 ppm Mg2+ +
Ba2+ and 12:900 ppm Zn2+ + Mg2+. Decrease %F = 22.54 and ethanol
yield 6.18 % (v/v) was obtained in medium containing 12:4 ppm Zn2+
+ Ba2+. In media containing the individual ions, increased %F =
27.94 and 26.03 were recorded for media containing 700 ppm Mg2+
and 2 ppm Ba2+ , with ethanol yield of 7.88% (v/v) and 7.62% (v/v)
respectively. Reduced %F and ethanol yield was observed for 10 ppm
Zn2+ and 4 ppm Ba2+ media. The impact of Ba2+ at 1 and 2 ppm was
significant.
Abstract: 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.
Abstract: A process of conversion of flour from three varieties of cassava, namely Odongbo, ofege and TMS30752 to ethanol using α-amylase locally sourced from germinated unhusked paddy rice and yeast isolated from palm wine was developed. It involves the germination of paddy rice for a period of 15days to produce α-amylase for starch hydrolysis and isolation of yeast from palm wine for fermentation. The results showed that optimum amylase yield of “ofada” rice paddy was at 6th day germination which was 576.9ml/g. Ethanol yield for TMS30572 (440.3%) was significantly higher than “Odongbo” (160.2%) and “Ofege’’ (115.1%), Sugar conversion efficiency were 311.0%v/v, 268.2%v/v and 186.84%v/v for TMS30572, “Odongbo” and “Ofege” respectively. The ethanol boiling points were 78oC, 76oC and 80oC for TMS30572, “Odongbo” and “Ofege” respectively. This study showed that cassava varieties affects quality of ethanol produced and germination of “ofada” rice for 6 days ensures optimum production of crude amylase enzyme.
Abstract: 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.
Abstract: Coproduction of fructose and ethanol from dates extract by a glucose-selective S. cerevisiae ATCC 36859 strain has been studied. Various initial sugar concentrations (i.e., 131.4, 315.3, 408.2, and 500.0 g/l) have been tested. The fermentation experiments were performed in a water shaker bath at 30°C and 120 rpm. The results showed that highest yields of fructose (95.0%) and ethanol (72.8%) were achieved for the 131.4 g/l concentration. Increasing the initial concentration to 315.3 g/l resulted in lower yields of fructose (82.2%) and ethanol (61.0%). However, further increase to 408.2 g/l increased the fructose yield (97.5%) at the expense of ethanol yield (42.0%) due to probable substrate inhibitions that resulted in lower glucose conversion. At 500 g initial sugar/l the growth rate of ATCC 36859 was highly inhibited.
Abstract: 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.
Abstract: The microbial production of ethanol from biodiesel¬derived crude glycerol by Enterobacter aerogenes TISTR1468, under micro-aerobic and anaerobic conditions, was investigated. The experimental results showed that micro-aerobic conditions were more favorable for cellular growth (4.0 g/L DCW), ethanol production (20.7 g/L) as well as the ethanol yield (0.47 g/g glycerol) than anaerobic conditions (1.2 g/L DCW, 6.3 g/L ethanol and 0.72 g/g glycerol, respectively). Crude glycerol (100 g/L) was consumed completely with the rate of 1.80 g/L/h. Two-stage fermentation (combination of micro-aerobic and anaerobic condition) exhibited higher ethanol production (24.5 g/L) than using one-stage fermentation (either micro-aerobic or anaerobic condition. The two- stage configuration, exhibited slightly higher crude glycerol consumption rate (1.81 g/L/h), as well as ethanol yield (0.56 g/g) than the one-stage configuration. Therefore, two-stage process was selected for ethanol production from E. aerogenes TISTR1468 in scale-up studies.
Abstract: The dilute acid pretreatment and enzymatic
saccharification of lignocellulosic substrate, cogon grass (Imperata
cylindrical, L.) was optimized prior ethanol fermentation using
simultaneous saccharification and fermentation (SSF) method. The
optimum pretreatment conditions, temperature, sulfuric acid
concentration, and reaction time were evaluated by determining the
maximum sugar yield at constant enzyme loading. Cogon grass, at
10% w/v substrate loading, has optimum pretreatment conditions of
126°C, 0.6% v/v H2SO4, and 20min reaction time. These
pretreatment conditions were used to optimize enzymatic
saccharification using different enzyme combinations. The maximum
saccharification yield of 36.68mg/mL (71.29% reducing sugar) was
obtained using 25FPU/g-cellulose cellulase complex combined with
1.1% w/w of cellobiase, ß-glucosidase, and 0.225% w/w of
hemicellulase complex, after 96 hours of saccharification. Using the
optimum pretreatment and saccharification conditions, SSF of treated
substrates was done at 37°C for 120 hours using industrial yeast
strain HBY3, Saccharomyces cerevisiae. The ethanol yield for cogon
grass at 4% w/w loading was 9.11g/L with 5.74mg/mL total residual
sugar.
Abstract: 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.