• Korean Chemical Engineering Research
• /
• v.52 no.1
• /
• pp.45-51
• /
• 2014

This is to study the effects of various pretreatment methods of agricultural residue, corn stover, and to compare the feature and pros and cons of each method including dilute sulfuric acid (DSA), soaking in aqueous ammonia (SAA), and ammonia recycle percolation (ARP). In order to convert corn stover to ethanol, various pretreatments followed by simultaneous saccharification and co-fermentation (SSCF) were tested and evaluated in terms of ethanol yield. With 3%, w/w of glucan loading using ARP-, DSA-, and SAA-treated solids, SSCFs using recombinant E. coli strain (ATCC$^{(R)}$ 55124) with commercial enzymes (15 FPU of Spezyme CP/g-glucan and 30 CBU/g-glucan enzyme loading) were tested. In the SSCF tests, 87, 90, and 78% of theoretical maximum ethanol yield were observed using ARP-, DSA-, and SAA-treated solids, respectively, which were 69, 58, and 74% on the basis of total carbohydrates (glucan + xylan) in the untreated corn stover. Ethanol yield of SAA-treated solid was higher than those of ARP- and DSA-treated solids. In addition, SSCF test using treated solids plus pretreated hydrolysate indicated that the DSA-treated hydrolysate showed the strongest inhibition effect on the KO11 strain, whereas the ARP-treated hydrolysate was found to have the second strongest inhibition effect. Bioconversion scheme using SAA pretreatment and SSCF can make the downstream process simple, which is suggested to produce ethanol economically because utilization of hemicellulose in the hydrolysate is not necessary.

• Journal of Life Science
• /
• v.23 no.10
• /
• pp.1192-1198
• /
• 2013

To improve yeast strains for bioethanol production, yeasts with ethanol tolerance, thermotolerance, and ${\beta}$-1,3-glucanase activity were bred using yeast genome shuffling. Saccharomyces cerevisiae $BY4742{\Delta}exg1$/pAInu-exgA, which has extracellular ${\beta}$-1,3-glucanase activity, and the Aspergillus oryzae and S. cerevisiae YKY020 strains, which exhibit ethanol tolerance and thermotolerance, were fused by yeast protoplast fusion. Following cell fusion, four candidate cells (No. 3, 9, 11, and 12 strains) showing thermotolerance at $40^{\circ}C$ were selected, and their ethanol tolerance (7% ethanol concentration) and ${\beta}$-1,3-glucanase activity were subsequently analyzed. All the phenotypes of the two parent cells were simultaneously expressed in one (No. 11) of the four candidate cells, and this strain was called BYK-F11. The BYK-F11 fused cell showed enhanced cell growth, ethanol tolerance, ${\beta}$-1,3-glucanase activity, and ethanol productivity compared with the $BY4742{\Delta}exg1$/pAInu-exgA and YKY020 strains. The results prove that a new yeast strain with different characters and the same mating type can be easily bred by protoplast fusion of yeasts.

• Journal of Life Science
• /
• v.23 no.7
• /
• pp.863-868
• /
• 2013

The XylP gene, which encodes endoxylanase in Bacillus sp. HY-20, was subcloned, and two expression plasmids, pG-xylP and pGMF-xylP were constructed. These plasmids, which contain different signal sequences, XylP s.s and $MF{\alpha}_{opt}$ s.s, respectively, for the secretory expression of endoxylanase, were transformed into Saccharomyces cerevisiae SEY2102 and FY833, respectively. The recombinant endoxylanases were successfully expressed, with a total activity range of 23.7-70.1 unit/ml according to the expression system and host strain. The endoxylanase activity in SEY2102/pGMF-xylP reached a maximum of 88.1 unit/ml in baffled flask culture. Most of the recombinant endoxylanase was efficiently secreted in the extracellular fraction, and the $MF{\alpha}_{opt}$ s.s was more efficient for secreting endoxylanase in yeast than the XylP s.s. Therefore, the expression system developed in this study produces large extracellular amounts of endoxylanase using S. cerevisiae as the host strain, and it could be used in bioethanol production and industrial applications.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.3
• /
• pp.390-399
• /
• 2012

Endoglucanase gene egIV was cloned from Trichoderma viride AS 3.3711, an important cellulose-producing fungus, by using an RT-PCR protocol. The egIV cDNA is 1,297 bp in length and contains a 1,035 bp open reading frame encoding a 344 amino acid protein with an estimated molecular mass of 35.5 kDa and isoelectronic point (pI) of 5.29. The expression of gene egIV in T. viride AS 3.3711 could be induced by sucrose, corn straw, carboxymethylcellulose (CMC), or microcrystalline cellulose, but especially by CMC. The transcripts of egIV were regulated under these substrates, but the expression level of the egIV gene could be inhibited by glucose and fructose. Three recombinant vectors, pYES2-xegIV, $pYES2M{\alpha}$-egIV, and $pYES2M{\alpha}$-xegIV, were constructed to express the egIV gene in Saccharomyces cerevisiae H158. The CMCase activity of yeast transformants $IpYES2M{\alpha}$-xegIV was higher than that of transformant IpYES2-xegIV or $IpYES2M{\alpha}$-egIV, with the highest activity of 0.13 U/ml at induction for 48 h, illustrating that the modified egIV gene could enhance CMCase activity and that $MF{\alpha}$ signal peptide from S. cerevisiae could regulate exogenous gene expression more effectively in S. cerevisiae. The recombinant EGIV enzyme was stable at pH 3.5 to 7.5 and temperature of $35^{\circ}C$ to $65^{\circ}C$. The optimal reaction condition for EGIV enzyme activity was at the temperature of $55^{\circ}C$, pH of 5.0, 0.75 mM $Ba^{2+}$, and using CMC as substrate. Under these conditions, the highest activity of EGIV enzyme in transformant $IpYES2M{\alpha}$-xegIV was 0.18 U/ml. These properties would provide technical parameters for utilizing cellulose in industrial bioethanol production.

• Korean Chemical Engineering Research
• /
• v.46 no.5
• /
• pp.977-982
• /
• 2008

The production of ethanol by microbial fermentation as an alternative energy source has been of interest because of increasing oil price. Saccharomyces cerevisiae and Zymomonas mobilis are two of the most widely used ethanol producers. In this study, characteristics of ethanol fermentation by Saccharomyces cerevisiae CHY1077 and Zymomonas mobilis CHZ2501 was compared. Brown rice, naked barley, and cassava were selected as representatives of the starch-based raw materials commercially available for ethanol production. The volumetric ethanol productivities by Saccharomyces cerevisiae from brown rice, naked barley and cassava were $0.68g/l{\cdot}h$, $1.03g/l{\cdot}h$ and $1.28g/l{\cdot}h$ respectively. But for the Zymomonas mobilis, $2.19g/l{\cdot}h$(brown rice), $2.60g/l{\cdot}h$(naked barley) and $3.12g/l{\cdot}h$(cassava) were obtained. Zymomonas mobilis was more efficient strain for ethanol production than S. cerevisiae.

• Journal of Life Science
• /
• v.27 no.12
• /
• pp.1403-1409
• /
• 2017

In this study, the xylitol dehydrogenase (XYL2) gene was expressed in Saccharomyces cerevisiae as a host cell for ease of use in the degradation of lignocellulosic biomass (xylose). To select suitable expression systems for the S.XYL2 gene from S. cerevisiae and the P.XYL2 gene from Pichia stipitis, $pGMF{\alpha}-S.XYL2$, $pGMF{\alpha}-P.XYL2$, $pAMF{\alpha}-S.XYL2$ and $pAMF{\alpha}-P.XYL2$ plasmids with the GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence was also connected to each promoter to allow secretion. Each plasmid was transformed into S. cerevisiae $SEY2102{\Delta}trp1$ strain and the xylitol dehydrogenase activity was investigated. The GAL10 promoter proved more suitable than the ADH1 promoter for expression of the XYL2 gene, and the xylitol dehydrogenase activity from P. stipitis was twice that from S. cerevisiae. The xylitol dehydrogenase showed $NAD^+$-dependent activity and about 77% of the recombinant xylitol dehydrogenase was secreted into the periplasmic space of the $SEY2102{\Delta}trp1/pGMF{\alpha}-P.XYL2$ strain. The xylitol dehydrogenase activity was increased by up to 41% when a glucose/xylose mixture was supplied as a carbon source, rather than glucose alone. The expression system and culture conditions optimized in this study resulted in large amounts of xylitol dehydrogenase using S. cerevisiae as the host strain, indicating the potential of this expression system for use in bioethanol production and industrial applications.

• Applied Chemistry for Engineering
• /
• v.22 no.5
• /
• pp.562-565
• /
• 2011

In this study, we developed a bioprocess using Clostridium ljungdahlii as a biological catalyst to produce bio-ethanol, and the effect of pH on microbial growth and ethanol production was investigated. From the results of fermentation at various initial pH condition without pH control, pH of fermentation broth decreased to 4.5 within 24 h due to accumulation of by-product acetic acid and both microbial growth and ethanol production were stopped. The experimental result of initial pH 8 showed the highest microbial growth and ethanol production (0.53 g/L), since the pH drop was relatively slow. From the experiment of pH 7 maintained fermentation using pH controllable bioreactor, the maximum cell dry weight of 1.65 g/L and the maximum ethanol concentration of 1.43 g/L were obtained within 24 h. In conclusion, the C. ljungdahlii growth was enhanced by pH maintenance of neutral range, and the ethanol production was also enhanced based on the growth-associated ethanol production characteristics of C. ljungdahlii.

• Korean Journal of Weed Science
• /
• v.30 no.4
• /
• pp.340-360
• /
• 2010

The depletion of fossil fuels, ecological problems associated with $CO_2$ emissions climate change, growing world population, and future energy supplies are forcing the development of alternative resources for energy (heat and electricity), transport fuels and chemicals: the replacement of fossil resources with $CO_2$ neutral biomass. Several options exist to cover energy supplies of the future, including solar, wind, and water power; however, chemical carbon source can get from biomass only. When used in combination with environmental friend production and processing technology, the use of biomass can be seen as a sustainable alternative to conventional chemical feedstocks. The biorefinery concept is analogous to today's petroleum refinery, which produce multiple fuels and chemical products from petroleum. A biorefinery is a facility that integrates biomass conversion processes and equipment to produce fuels, power, and value-added chemicals from biomass. Biorefinery is the co-production of a spectrum of bio-based products (food, feed, materials, and chemicals) and energy (fuels, power, and heat) from biomass [definition IEA Bioenergy Task 42]. By producing multiple products, a biorefinery takes advantage of the various components in biomass and their intermediates therefore maximizing the value derived from the biomass feedstocks. A biorefinery could, for example, produce one or several low-volume, but high-value, chemical or nutraceutical products and a low-value, but high-volume liquid transportation fuel such as biodiesel or bioethanol. Future biorefinery may play a major role in producing chemicals and materials as a bridge between agriculture and chemistry that are traditionally produced from petroleum. Industrial biotechnology is expected to significantly complement or replace the current petroleum-based industry and to play an important role.

• KSBB Journal
• /
• v.24 no.5
• /
• pp.439-445
• /
• 2009

The process for ethanol production requires lignocellulosic biomass to be hydrolyzed to generate monomeric sugars for the fermentation. During hydrolysis step, a monomeric sugars and a broad range of inhibitory compounds (furan derivatives, weak acids, phenolics) are formed and released. In this study, we investigated the effects of inhibitory compounds on the fermentative performance of Saccharomyces cerevisiae K35 and Pichia stipitis KCCM 12009 in ethanol production, two yeast strains were fermented in the synthetic medium including six inhibitory compounds such as 5-hydroxymethylfurfura (5-HMF), furfural, acetic acid, syringaldehyde, vanillic acid and syringic acid. Ethanol of over 40 g/L was produced by two yeast strains in the absence of inhibitory compounds, respectively. Most inhibitory compounds except acetic acid had a little effect on the ethanol production, but acetic acid showed high inhibition effect on the cell growth and ethanol production.

• Journal of the Korean Wood Science and Technology
• /
• v.37 no.3
• /
• pp.255-264
• /
• 2009

To evaluate the effects of chemical pretreatments of lignocellulosic biomass on enzymatic hydrolysis process, Populus euramericana was pretreated for 1 hr with 1% sulfuric acid ($H_2SO_4$) at $150^{\circ}C$ and 1% sodium hydroxide (NaOH) at $160^{\circ}C$, respectively. Before the enzymatic hydrolysis, each pretreated sample was subjected to drying process and thus finally divided into four subgroups; dried or non-dried acid pretreated samples and dried or non-dried alkali pretreated samples and chemical and physical properties of them were analyzed. Biomass degradation by acid pretreatment was determined to 6% higher compared to alkali pretreatment. By the action of acid ca. 24.5% of biomass was dissolved into solution, while alkali degraded ca. 18.6% of biomass. However, reverse results were observed in delignification rates, in which alkali pretreatment released 2% more lignin fragment from biomass to the solution than acid pretreatment. Unexpectedly, samples after both pretreatments were determined to somewhat higher crystallinity than untreated samples. This result may be explained by selective disrupture of amorphous region in cellulose during pretreatments, thus the cellulose crystallinity seems to be accumulated in the pretreated samples. SEM images revealed that pretreated samples showed relative rough and partly cracked surfaces due to the decomposition of components, but the image of acid pretreated samples which were dried was similar to that of the control. In pore size distribution, dried acid pretreated samples were similar to the control, while that in alkali pretreated samples was gradually increased as pore diameter increased. The pore volume which increased by acid pretreatment rapidly decreased by drying process. Alkali pretreatment was much more effective on enzymatic digestibility than acid pretreatment. The sample after alkali pretreatment was enzymatically hydrolyzed up to 45.8%, while only 26.9% of acid pretreated sample was digested at the same condition. The high digestibility of the sample was also influenced to the yields of monomeric sugars during enzymatic hydrolysis. In addition, drying process of pretreated samples affected detrimentally not only to digestibility but also to the yields of monomeric sugars.