• Journal of Microbiology and Biotechnology
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• v.4 no.3
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• pp.215-221
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• 1994

Two strains of yeast (RA-74-2 and RA-912) showing superior fermenting ability at a high temperature were isolated from soils and wastewaters by an enrichment culture method. Based on the morphological and physiological charateristics, the two strains were identified as Saccharomyces cerevisiae and Kluyveromyces marxianus, respectively. RA-74-2 was able to grow upto $43^{\circ}C$ and sustain similar fermenting ability in the temperatures range from 30 to $40^{\circ}C$. In addition, the sugar- and ethanol-tolerance of RA-74-2 were 30% (w/v) glucose and 10% (v/v) ethanol, which appeared to be higher than those of nine other industrial yeast strains currently being used in the alcohol factories. The thermotolerant ethanol fermenting yeast RA-912 showed identical growth in the temperatures range from 35 to $45^{\circ}C$ and was resistant to various heavy metals. The quality and quantity of byproducts of the isolated yeast strains in fermentation broth after fermentation at $40^{\circ}C$ and $45^{\circ}C$ were similiar with those obtained at $30^{\circ}C$. These results show that RA-74-2 can be adopted for the ethanol fermentation process where the expenses for cooling system is significant, and suggest that RA-912 may be applied in either SSF(simultaneous saccharification and fermentation) or Flash-fermentation process and RA-912 may be used as a gene donor for the development of thermotolerant ethanol-fermenting yeasts.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.718-726
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• 2010

A selected fungal strain, for production of the raw-starchdigesting enzyme by solid-state fermentation, was improved by two repeated sequential exposures to ${\gamma}$-irradiation of $Co^{60}$, ultraviolet, and four repeated treatments with Nmethyl-N'-nitrosoguanidine. The mutant strain Aspergillus sp. XN15 was chosen after a rigorous screening process, with its production of the raw-starch-digesting enzyme being twice that of usual wild varieties cultured under preoptimized conditions and in an unsupplemented medium. After 17 successive subculturings, the enzyme production of the mutant was stable. Optimal conditions for the production of the enzyme by solid-state fermentation, using wheat bran as the substrate, were accomplished for the mutant Aspergillus sp. XN15. With the optimal fermentation conditions, and a solid medium supplemented with nitrogen sources of 1% urea and 1% $NH_4NO_3$, 2.5 mM $CoSO_4$, 0.05% (v/w) Tween 80, and 1% glucose, the mutant Aspergillus sp. XN15 produced the raw-starch-digesting enzyme in quantities 19.4 times greater than a typical wild variety. Finally, XN15, through simultaneous saccharification and fermentation of a raw rice corn starch slurry, produced a high level of ethanol with $Y_{p/s}$ of 0.47 g/g.

• Molecules and Cells
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• v.28 no.4
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• pp.369-373
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• 2009

Heterologous secretory expression of endoglucanase E (Clostridium thermocellum) and ${\beta}$-glucosidase 1 (Saccharomycopsis fibuligera) was achieved in Saccharomyces cerevisiae fermentation cultures as an ${\alpha}$-mating factor signal peptide fusion, based on the native enzyme coding sequence. Ethanol production depends on simultaneous saccharification of cellulose to glucose and fermentation of glucose to ethanol by a recombinant yeast strain as a microbial biocatalyst. Recombinant yeast strain expressing endoglucanase and ${\beta}$-glucosidase was able to produce ethanol from ${\beta}$-glucan, CMC and acid swollen cellulose. This indicates that the resultant yeast strain of this study acts efficiently as a whole cell biocatalyst.

• Korean Chemical Engineering Research
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• v.54 no.4
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• pp.494-500
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• 2016

In order to remove acetyl group from yellow poplar, deacetylation was performed using sodium hydroxide (NaOH) prior to oxalic acid pretreatment. During the deacetylation ($60^{\circ}C$ for 80 min, 0.8% NaOH), most of the acetyl group were removed from hemicellulose. Simultaneous saccharification and fermentation (SSF) and semi-SSF were carried out based on solid loading (10, 12.5, 15%) of deacetylated biomass and pre-hydrolysis with enzymes (0, 6, 12, 24 h). The highest ethanol was obtained as 26.73 g/L after 120 h when 10% of biomass was used for SSF. It is corresponding to 88.41% of theoretical ethanol yield. At the 12.5% and 15% of biomass loading, the highest ethanol was obtained from 6 h pre-hydrolysis. It was 32.34 g/L and 27.15 g/L, respectively, and corresponding to ethanol yield of 85.58 and 59.87%. In order to remove fermentation inhibitors from hydrolysates, overliming was performed using calcium hydroxide ($Ca(OH)_2$). The highest ethanol was 5.28 g/L after 72 h of fermentation.

• Journal of the Korean Wood Science and Technology
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• v.41 no.6
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• pp.507-514
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• 2013

In this study, oxalic acid pretreatment of empty fruit bunch (EFB) was performed at different pretreatment temperatures. Also, we evaluated oxalic acid recovery from hydrolysate by electrodialysis. The fermentable sugar concentration in hydrolysate was high at more than $20g/{\ell}$, when pretreatment was carried out at $150^{\circ}C$. At the same time, ethanol production was $3.78g/{\ell}$ after 72 h which correspond to the ethanol yield of 0.21 g/g. On the other hydrolysate (160, $170^{\circ}C$), fermentable sugar was not consumed by Pichia stipitis during fermentation. Most of the oxalic acid was recovered and some of the fermentation inhibitors were removed by electrodialysis. For the electrodialysis treated hydrolysate, ethanol production was increased compared to the original hydrolysate. The highest ethanol production was $5.38g/{\ell}$ after 24 h which correspond to the yield of 0.33 g/g. The ethanol production by simultaneous saccharification and fermentation (SSF) under all pretreatment conditions was more than $15g/{\ell}$ after 96 h. The highest ethanol production was $20.54g/{\ell}$, when pretreatment was performed at $170^{\circ}C$. In particular, ethanol production was increased, when electrodialysis treated hydrolysate was used for SSF.

• Microbiology and Biotechnology Letters
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• v.18 no.5
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• pp.506-510
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• 1990

The Takju brewing of raw rice starch was carried out by the simultaneous saccharification- fermentation using Rhizopus sp. koji and yeast, and compared with the Takju mash brewed by the conventional method. Rhitopus koji was prepared with uncooked rice for Takju brewing without cooking of rice starch. Alcohol concentration of Takju mash brewed with uncooked rice was slightly higher of 1.8% than that with cooked one. Amino acid contents was almost double and fuse1 oil contents was lower in uncooked brewing. The Takju mash prepared after fermentation without cooking of rice had a characteristic odor of raw material and a good quality of taste.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1401-1405
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• 2012

A thermotolerant Saccharomyces cerevisiae mutant strain, TT6, was constructed after multi-parental hybridization of five mutant strains obtained by UV or NTG treatment of the original strain, S. cerevisiae KV1. When incubated at $40^{\circ}C$ in YPD broth, TT6 began to grow exponentially in 10 h, but KV1 did not show any noticeable growth even after 22 h. The thermotolerant growth of TT6 was confirmed by serial dilution assay at $42^{\circ}C$; TT6 grew at a cell concentration ($10^{-5}$) 10,000 times lower than that of KV1 ($10^{-1}$). Whereas ethanol production from YP containing 23% (w/v) glucose by KV1 decreased with increasing temperature from $30^{\circ}C$ to $36^{\circ}C$, ethanol production by TT6 did not decrease at temperatures up to $37^{\circ}C$. When TT6 was tested for ethanol production at $36^{\circ}C$ by simultaneous saccharification and fermentation (SSF) from 23% corn, 24 h of fermentation time or 50% of the glucoamylase dose was saved when compared with KV1 at $30^{\circ}C$. The ethanol yield from corn by SSF with TT6 at $36^{\circ}C$ was 91.7% of the theoretical yield, whereas that of KV1 at $30^{\circ}C$ was 90.6%.

• KSBB Journal
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• v.25 no.5
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• pp.471-477
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• 2010

In order to improve bio-ethanol productivity by various cultivation methods in this paper, the culture modes using food wastes, such as batch culture, high-cell-density fermentation, SSF (simultaneous saccharification and fermentation) by fill & draw, continuous culture by fill & draw were performed and their productivities were compared. SSFs by fill & draw were performed by continuous decompression using 1 L evaporator system, and by 10 L bioreactor without decompression. In addition, the continuous cultures by fill & draw mode using SFW (saccharafied food wastes) medium were performed by changes of 40% culture broth with intervals of 12 h (0.03 $h^{-1}$), 6 h (0.07 $h^{-1}$), 3 h (0.13 $h^{-1}$). Consequently, productivities of bio-ethanol were 2.52 g/L-h and 1.30 g/L-h in batch culture and high- cell-density fermentation, respectively. The productivities of SSF by fill & draw showed 2.24 g/L-h and 2.03 g/L-h in continuous decompression with 1 L evaporator and 10 L bioreactor without decompression, respectively. Also, the productivities in continuous culture by fill & draw modes showed 2.02 g/L-h, 4.07 g/L-h and 6.25 g/L-h by medium change with intervals of 12 h, 6 h, and 3 h, respectively. In conclusion, the highest ethanol productivity was obtained in the continuous culture mode by fill & draw with dilution rate of 0.13 $h^{-1}$.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.430-435
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• 1987

A chitin-immobilized enzyme amyloglucosidase(AMG) and a bacterium Zymomonas mobilis were coentrapped in alginate gel beads. Ethanol production was performed in a packed bed column reactor in a simultaneous saccharification and fermentation(SSF) mode using liquefied sago starch as a substrate. It was found that this process eliminated product inhibition and reverse reaction of glucose enhancing the rate of saccharification and ethanol production. At a low dilution rate of D = 0.11 hr$^{-1}$, the steady-state ethanol concentration was 46.0g/$m\ell$ (96.8 % of theoretical yield). The maximum ethanol productivity was 17.7g/$m\ell$, h at D = 0.83 hr$^{-1}$ when the calculation was based on the total working volume. The continuous production of ethanol was maintained stably over 40 days without problems in this reactor system.

• Journal of the Korean Wood Science and Technology
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• v.37 no.4
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• pp.397-405
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• 2009

In this study, we investigated the potential of producing bioethanol from Liriodendron tulipifera by using oxalic acid pretreatment. Amounts of fermentable sugars, mostly xylose and glucose, in the liquid fraction (hydrolysate) was $40.22g/{\ell}$ after the biomass was pretreated with 0.037 g/g of oxalic acid for 20 minutes at $160^{\circ}C$. Production amounts of ethanol was $8.6g/{\ell}$ from the 72 hours of simultaneous saccharification and fermentation (SSF) on solid fraction of the pretreated sample. At the same condition, when the reaction time increased to 40 minutes, $32.66g/{\ell}$ of fermentable sugars in the hydrolysate and $9.5g/{\ell}$ of ethanol was produced from the process of pretreatment and SSF. As a result of analyzing the fermentation inhibitors, such as acetic acid, 5-HMF, furfural and total phenolic compounds, as the reaction time increased, the amount of the fermentation inhibitors in the hydrolysate increased. Production of the fermentation inhibitors was more affected by initial concentration of oxalic acid rather than reaction time. $3.39{\sim}5.78g/{\ell}$ of acetic acid was produced by pretreatment with 0.013 g/g of oxalic acid, and the amount of furfural produced by decomposition of xylose was 2~3 times higher than the amount of 5-HMF produced by decomposition of glucose. All the hydrolysates contained more than $5g/{\ell}$ of total phenols considered as the degradation product of lignin. Therefore, by analyzing the amount of fermentable sugars and fermentation inhibitors in the hydrolysate, and producing ethanol from SSF of solid fraction of the pretreated sample, the biomass pretreated with 0.037 g/g of oxalic acid for 20 minutes at $160^{\circ}C$ can be expected to produce the most ethanol.