• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.145-149
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• 2016

Thermal acid hydrolysis pretreatment of Kappaphycus alvarezii was carried out with 12% (w/v) seaweed slurry and 180 mM H₂SO₄ at 140°C for 5 min. Utility of the thermotolerant yeast Kluyveromyces marxianus KCTC7150 was evaluated with respect to cell growth and ethanol fermentation at 40°C was close to optimal for enzymatic hydrolysis. This could lead to the integration of both the saccharification and fermentation processes. The levels of ethanol production by simultaneous saccharification and fermentation (SSF) with non-adapted and adapted K. marxianus KCTC7150 were 9.1 g/l with an ethanol yield (Y_EtOH) of 0.24 and 10.2 g/l with an ethanol yield (Y_EtOH) of 0.27 at 156 h, respectively. The two-phase SSF process was employed in this study to improve the efficiency of ethanol fermentation. Adapted K. marxianus KCTC7150 using the two-phase SSF process produced 13.5 g/l with an ethanol yield (Y_EtOH) of 0.35 at 96 h. Development of the two-phase SSF process could enhance the overall ethanol fermentation yields of the seaweed K. alvarezii.

• Korean Journal of Soil Science and Fertilizer
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• v.37 no.1
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• pp.46-53
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• 2004

In this study, we compared the levels of methylotrophic bacterial community diversity in the leaf, stem, grain, root and rhizosphere soil sainples of four rice cultivars collected from three regions of Korea. Thirty five pigmented and five non-pigmented isolates showing characteristic growth on methanul were obtained. When phylotypes were defined by performing numerical analysis of 42 characteristics, four distinct clusters were formed. While two clusters, I and IV diverged on the basis of nitrate and nitrite reduction, other two clusters, comprising only pink pigmented colonies, diverged on the basis of cellulase activity. Out of the two reference strains used in the analysis, Methyhbacterium extorquens AM1 diverged from all the clusters and M. fujisawaense KACC 10744 grouped under cluster III. All the isolates were positive for urease, oxidase, catalase and pectinase activity and negative for indole production, MR and VP test, $H_2S$ production, starch, and casein hydrolysis. No clusters were found to possess thermotolerant isolates, as no growth of the isolates was observed at $45^{\circ}C$. Two strains in cluster I were found to possess gelatin hydrolysis and methane utilizing properties respectively. Most of the isolates in all the four clusters utilized monosaccliarides, disaccharide and polyols as carbon source. Six isolates showed considerable nitrogenase activity ranging from 86.2 to $809.9nmol\;C_2H_4\;h^{-1}\;mg^{-1}$ protein.

• Microbiology and Biotechnology Letters
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• v.25 no.4
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• pp.386-390
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• 1997

The characteristics of alcohol dehydrogenase (ADH, EC 1.1.1.1, alcohol:NAD oxidoreductase) of thermotolerant alcohol-producing yeasts, Saccharomyces cerevisiae RA-74-2 and Kluyveromyces marxianus RA-912, were compared with that of mesophilic S. cerevisiae D, an industrial strain. Under anaerobic culture condition, both S. cerevisiae RA-74-2 and D had similar level of ADH activity at 30$\circ$C, and the activity of S. cerevisiae RA-74-2 at 37$\circ$C was the same level at 30$\circ$C. However, the level of ADH activity of S. cerevisiae D at 37$\circ$C decreased about 70% of that at 30$\circ$C. The level of enzyme activity of K. marxianus RA-912, which showed lower alcohol productivity than S. cerevisiae RA-74-2 and D, was about 43% of those strains at 30$\circ$C, and decreased somewhat at 37$\circ$C. The results showed a good correlation between the alcohol productivities and the level of ADH activities of these strains grown at 30$\circ$C and 37$\circ$C. And the higher heat stability of ADH of S. cerevisiae RA-74-2 than that of S. cerevisiae D seemed to reflect the ability of high temperature fermentation. Despite of its fermentation ability even at 45$\circ$C, however, the ADH of K. marxianus RA-912 showed lower heat stability than that of S. cerevisiae D. Both S. cerevisiae RA-74-2 and D showed similar patterns of two bands of ADH isozyme, and the low band of S. cerevisiae RA-74-2 moved slightly faster than that of S. cerevisiae D. The staining intensity of the bands of S. cerevisiae D at 37$\circ$C was weaker than those at 30$\circ$C. However, S. cerevisiae RA-74-2 showed no differences in total intensity of the bands of 30$\circ$C and 37$\circ$C. As the patterns of cellular proteins and ADH isozyme of K. marxianus RA-912 were different from S. cerevisiae RA-74-2 and D, K. marxianus might have its own characteristic ADH system.

• Microbiology and Biotechnology Letters
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• v.11 no.1
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• pp.5-13
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• 1983

This experiment was carried out to elucidate the thermotolerant properties of a thermophilic bacterium which isolated from soils of the hot springs area and selected for the $\beta$-galactosidase production. Biochemical and physiological characteristics of this strain were studied, including the investigation on the fatty acid composition for its neutral fats. The results obtained were summarized as follows. 1. This bacterium was identified as a strain belong to the genus Thermus. 2. Optimal temperature and pH for growth of this strain were $65^{\circ}C$ and pH 6.5 respectively, and it was found to be an absolute thermophilic bacterium which could not grow at 37$^{\circ}C$. 3. No growth was obtained in the medium which contained more than 1.0% of sodium chloride. 4. The tolerable concentration of antobiotics were 10$\mu\textrm{g}$ of penicillin G per $m\ell$ of medium and 0.5$\mu\textrm{g}$ of chloramphenicol per $m\ell$ respectively 5. This strain had autotrophilic requirements for calcium-pantothenate and pyridoxine-HCO as an-essential factor and for niacin as a stimulative factor. 6. Fatty acid composition of neutral fats of the strain was palmitic acid. 60.20%; lauric acid, 11.8%; myristic acid, 7.56%, behenic acid, 4.25%; capric acid, 1.77%; stearic acid, 2.13%; arachidic acid, 1.53%; and others unidentified, 10.7%.

• The Journal of Natural Sciences
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• v.16 no.1
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• pp.15-29
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• 2005

To produce a thermostable cyclodextrin by using thermotolerant cyclomaltodextrin glucanotransferase(CGTase), a thermophilic and alkalophilic bacterium isolate, designated B-13 showing the highest CGTase activity was isalated from natural sources and identified as Bacillus cereus B-13 based on the morphological and physiological characteristics, and 16S rRNA sequence. The maximal CGTase activity (130 U/ml) was obtained when Bacillus cereus B-13 was cultured in SYC medium containing 2.0% soluble starch, 1.0% yeast extracts, 1% corn steep liquor and 1% $Na_2CO_3$ (pH 8.5) at $50^{\circ}C$ for 24 h and about 80% of maximal activity was also showed in he culture broth of $60^{\circ}C$ for 18 h. Optimum reaction temperature and pH of the partial purified CGTase for soluble starch were $65^{\circ}C$ and pH 8.5-9.0 respectively. The partial purified CGTase were also stable below $80^{\circ}C$ and pH 5.0-10.0. When 1% soluble starch was digested with the partial purified CGTase, the yield of cyclodextrin was 49%.

• Korean Journal of Agricultural Science
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• v.9 no.1
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• pp.377-386
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• 1982

This experiment was carried out to elucidate the thermotolerant properties of a thermophilic bacterium which was isolated from soils of the hot springs area and selected for the ${\beta}$-galactosidase production. This bacterium was identified as a strain belong to the genus Thermus. Biochemical and physiological characteristics of this strain were studied, including the investigation of the fatty acid composition of its neutral fats. The results obtained were summarized as follows. 1. Optimal temperature and pH for growth of this strain were $65^{\circ}C$ and pH 6.5 respectively, and it was found to be an absolute thermophilic bacterium which could not grow at the temperature below $43^{\circ}C$. 2. No growth was obtained in the medium which contained more than 1.0% of sodium chloride. 3. The tolerable concentration of antibiotics were 10mg of penicillin G per ml of medium and 0.5mg of chloramphenicol per ml respectively. 4. This strain had auxotrophilic requirements for calcium-pantothenate and pyridoxin-HCl as an essential factor and for niacin as a stimulative factor. 5. Yellow pigment was released into the liquid culture of this strain, which showed maximum absorption at 420 nm. 6. Fatty acid composition of neutral fats of the strain was palmitic acid, 60.20%; lauric acid, 11.80%; myristic acid, 7.56%; behenic acid, 4.25%; capric acid, 1.77%; stearic acid, 2.13%; arachidic acid, 1.53%; and others unidentified, 10.7%.

• Journal of The Korean Society of Grassland and Forage Science
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• v.25 no.4
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• pp.267-274
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• 2005

To develop transgenic orchardgrass (Dactylis glomerata L.) resistant to high temperature, a thermptolerance gene, DgP23, was introduced into orchardgrass using Agrobacterium - mediated transformation method. PCR and Southern blot analyses using genomic DNA showed specific DNA band on agarose gel and hybridization signal on X- ray film in transgenic orchardgrass harboring the recombinant DgP23 gene, but not in the wild type and empty vector control plants. RT-PCR and Southern blot analyses using total RNA also showed specific DNA band and hybridization signal. Transgenic orchardgrass did not showed ny morphological aberration both in the green house and field cultivation. Thermotolerance of transgenic plants was not detected in laboratory test. but may detected in field test.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.2
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• pp.99-106
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• 2011

In order to investigate rice stem proteome in response to heat stress, rice plants were subjected to heat treatment at 42$^{\circ}C$ and total soluble proteins were extracted from stem tissues, and were fractionated with 15% PEG (poly ethylene glycol) and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). After staining of 2-DE gels, 46 of differentially expressed proteins were extracted, digested by trypsin, and subjected to matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Proteins were identified through database search by using peptide mass fingerprints. Among them, 10 proteins were successfully identified. Seven proteins were up- and 3 proteins were down-regulated, respectively. These proteins are involved in energy and metabolism, redox homeostasis, and mitochondrial small heat shock proteins. The identification of some novel proteins in the heat stress response provides new insights that can lead to a better understanding of the molecular basis of heat-sensitivity in plants, and also useful to molecular breeding of thermotolerant forage crops.

• Microbiology and Biotechnology Letters
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• v.31 no.1
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• pp.63-68
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• 2003

Saccharomyces cerevisiae KNU5377 is a constitutively thermotolerant, fermentative strain at high temperatures over 4$0^{\circ}C$. The exposure to 0.5 M NaCl caused S. cerevisiae KNU5377 to be lost its constitutive thermotolerance. Furthermore, the NaCl adaptation beyond 0.3 M during the overnight culture forced the strain-specific fermentation ability of S. cerevisiae KNU5377 to be disappeared. However, these phenomena did not occur in the reference, Saccharomyces cerevisiae ATCC24858. As a result, this adaptation led both strains to show the closely similar thermotolerance level and alcohol fermentation ability, implying the NaCl adaptation eliminated its strain-specific characteristics of S. cerevisiae KNU5377 Therefore it indicated that the superior intrinsic characteristics of S. cerevisiae KNU5377 must be related to the NaCl adaptation. On the other hand, the heat adaptation elevated alcohol productivity for both strains, but surprisingly did it for KNU5377 at the rate of two times higher than the reference's one; this suggests that KNU5377 possesses more efficient system enough to cause the difference. Consequently, these characteristics of S. cerevisiae KNU5377 must be interesting targets for further study to understand on how KNU5377 could acquire the constitutive thermotolerance and the outstanding fermentative capacity at high temperatures.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1636-1643
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• 2012

Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-${\beta}$-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at $65-70^{\circ}C$ with an optimal pH at 9-10 and retaining >80% activity at pH 9, $60^{\circ}C$ for 1 h. Xyn3F showed a $V_{max}$ of 2,327 IU/mg and $K_m$ of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.