• Journal of Life Science
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• v.15 no.6 s.73
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• pp.884-888
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• 2005

An agar-degrading bacterium was isolated from north-eastern sea of Jeju island and cultured in marine agar 2216 media. Biochemical and morphologicl characteristics and 165 rRNA gene revealed that isolated strain was member of Agarivorans genus, and named Agarivorans sp. JA-1. Agarase was produced as growth-related and expressed regardless of agar presence. Optimal pH was 8 at 50 mM Clycine-NaOH buffer, and activity was maximum at $40^{\circ}C$E Enzymatic activity was maintained over $80\%$ at $60^{\circ}C$t and $70\%$ at $80^{\circ}C$ which is thermotolerant. Hence isolated novel Agarivorans sp. JA-1 strain and its beta-agarase could be used for the production of functional oligosaccharide from agar in solution state.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.1011-1014
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• 2003

The objective of this work was to evaluate a thermotolerant yeast Issatchenkia orientalis DY252 as a microbial feed additive for ruminants. In the present study, the influence of volatile fatty acids (VFA) and temperature on oxygen uptake rate by I. orientalis DY 252 was investigated. It was evident that the oxygen uptake rate was decreased gradually as the VFA concentrations increased in a range of 30 to 120 mM. Although the oxygen uptake rate was not greatly affected by temperature in the range 37 to $43^{\circ}C$, a maximum value of $0.45mg\;O_2/g$ cell/ min was obtained at $39^{\circ}C$. With regard to the oxygen uptake rate by yeast, viability was found to be less important than the metabolic activity of yeast.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1178-1182
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• 2005

Survival of thermotolerant Lactobacillus reuteri KUB-AC5 in $20\%$ (w/v) skim milk was found to be $11.3\%$ after spray drying by using a pilot scale spray dryer with inlet temperature at $170^{\circ}C$ and outlet temperature at $85^{\circ}C$. The ability of dried cell to produce antimicrobial activity was not affected by the spray drying. The model system for predicting viability of spray-dried L. reuteri KUB-AC5 during long-term storage was established, based on the Arrhenius equation, and verified by experimental data, because the viability of cells during storage can be correlated with storage temperature. The viability during storage at $30^{\circ}C$ declined more rapidly than that storage at $4^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1230-1237
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• 2009

The selective plugging strategy of Microbial Enhanced Oil Recovery (MEOR) involves the use of microbes that grow and produce exopolymeric substances, which block the high permeability zones of an oil reservoir, thus allowing the water to flow through the low permeability zones leading to increase in oil recovery. Bacillus licheniformis TT33, a hot water spring isolate, is facultatively anaerobic, halotolerant, and thermotolerant. It produces EPS as well as biosurfactant and has a biofilm-forming ability. The viscosity of its cell-free supernatant is $120\;mPa{\cdot}s$ at $28^{\circ}C$. Its purified EPS contained 26% carbohydrate and 3% protein. Its biosurfactant reduced the surface tension of water from 72 to 34 mN/m. This strain gave $27.7{\pm}3.5%$ oil recovery in a sand pack column. Environmental scanning electron microscopy analysis showed bacterial growth and biofilm formation in the sand pack. Biochemical tests and Amplified Ribosomal DNA Restriction Analysis confirmed that the oil recovery obtained in the sand pack column was due to Bacillus licheniformis TT33.

• Journal of The Korean Society of Grassland and Forage Science
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• v.20 no.2
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• pp.97-102
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• 2000

To increase thennotolerance of forage crops, transgenic rice plants as a model for transformation of monocots were generated. A cDNA encoding the chloroplast-localized small heat shock protein (small HSP) of rice, Oshsp21, was introduced into rice plants via Agrobacterium-mediated gene transfer system. Calli induced from scutella were co-cultivated with a A. tumefaciens strain EHAlOl canying a plasmid, pIGhsp21. A large number of transgenic plants were regenerated on a medium containing hygromycin. Integration of Oshsp2l gene was confirmed by PCR and Southern blot analyses with genomic DNA. Northern blot and immunoblot analyses revealed that the Oshsp21 gene was constitutively expressed and accumulated as mature protein in transgenic plants. Effects of constitutive expression of the OshspZl on thermotolerance were first probed with the chlorophyll fluorescence. Results indicate that inactivation of electron transport reactions in photosystem I1 (PSII), were mitigated by constitutive expression of the Oshsp21. These results suggest that the chloroplast small HSP plays an important role in protecting photosynthetic machinery during heat stress. (Key words : Thermotolerance, Rice, Transgenic, cDNA)

• Journal of the Korea Organic Resources Recycling Association
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• v.12 no.3
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• pp.119-125
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• 2004

Sugar beet stillages were used as a substrate for the production of single cell protein by a thermotolerant yeast Candida rugosa. 3 Stillage substrates were nutritionally optimized for the better production of yeast biomass and for the reduction of COD. The addition of Phosphorus(P) was required for all stillages, but Nitrogen(N) only when the residual sugar remained. The addition of P increased the biomass production to 23-61%. The addition of N increased the biomass production only a little, but when added together with P increased to 90%. The COD decreased to 26-46% when P was added, but decreased to 85% when P was added together with N.

• BMB Reports
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• v.40 no.3
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• pp.333-340
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• 2007

Gene encoding cyclodextrin glycosyltransferase (CGTase), from thermotolerant Paenibacillus sp. T16 isolated from hot spring area in northern Thailand, was cloned and expressed in E. coli (JM109). The nucleotide sequences of both wild type and transformed CGTases consisted of 2139 bp open reading frame, 713 deduced amino acids residues with difference of 4 amino acid residues. The recombinant cells required 24 h culture time and a neutral pH for culture medium to produce compatible amount of CGTase compared to 72 h culture time and pH 10 for wild type. The recombinant and wild-type CGTases were purified by starch adsorption and phenyl sepharose column chromatography and characterized in parallel. Both enzymes showed molecular weight of 77 kDa and similar optimum pHs and temperatures with recombinant enzyme showing broader range. There were some significant difference in pH, temperature stability and kinetic parameters. The presence of high starch concentration resulted in higher thermostability in recombinant enzyme than the wild type. The recombinant enzyme was more stable at higher temperature and lower pH, with lower $K_m$ for coupling reaction using cellobiose and cyclodextrins as substrates.

• Mycobiology
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• v.33 no.2
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• pp.90-96
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• 2005

Sixty two out of the sixty four species of fungal isolates tested could produce both $exo-{\beta}1,4-gluconase\;(C_1)$ and $endo-{\beta}1,4-gluconase\;(C_x)$ on pure cellulose and rice straw as carbon source in Czapek's medium. Fifty-eight and fifteen species were able to grow at $25^{\circ}C$ and at $45^{\circ}C$, respectively. Eleven species could grow at both $25^{\circ}C$ and $45^{\circ}C$ while, four species appeared only at $45^{\circ}C$. The most cellulolytic species at $25^{\circ}C$ was Trichoderma koningii producing 1.164 $C_1$ (mg glucose/1 ml culture filtrate/1 hr) and 2.690 $C_x$ on pure cellulose, and 0.889 $C_1$, and 1.810 $C_x$ on rice straw, respectively. At $45^{\circ}C$, the most active thermotolerant species were Aspergillus terreus, followed by A. fumigatus. Talaromyces thermophilus was the highest active thermophilic species followed by Malbranchea sulfurea. Most of these species were also active in fermentation of rice straw at 25 and $45^{\circ}C$ (P<0.05). The most active ones were T. koningii, A. ochraceus and A. terreus, which produced 201.5, 193.1 and 188.1 mg crude protein/g dry straw, respectively.

• Journal of Microbiology and Biotechnology
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• v.25 no.1
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• pp.66-73
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• 2015

The pabS gene of Agaricus bisporus 02 encoding a putative PABA synthase was cloned, and then the recombinant protein was expressed in Escherichia coli BL21 under the control of the T7 promoter. The enzyme with an N-terminal GST tag or His tag, designated GST-AbADCS or His-AbADCS, was purified with glutathione Sepharose 4B or Ni Sepharose 6 Fast Flow. The enzyme was an aminodeoxychorismate synthase, and it was necessary to add with an aminodeoxychorismate lyase for synthesizing PABA. AbADCS has maximum activity at a temperature of approximately 25℃ and pH 8.0. Magnesium or manganese ions were necessary for the enzymatic activity. The Michaelis-Menten constant for chorismate was 0.12 mM, and 2.55 mM for glutamine. H₂O₂ did distinct damage on the activity of the enzyme, which could be slightly recovered by Hsp20. Sulfydryl reagents could remarkably promote its activity, suggesting that cysteine residues are essential for catalytic function.

• KSBB Journal
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• v.25 no.6
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• pp.565-571
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• 2010

To develop thermostable ethanol fermentative yeast strain for lignocellulosic simultaneous saccharification and fermentation, high ethanol producing yeast, Saccharomyces cerevisiae CHY1012 and thermostable yeast, Kluyveromyces marxianus CHY1703 were fused by protoplast fusion. The thermostable fusant, CHY1612 was identified as a Kluyveromyces marxianus by phenotypic and physiological characteristics, as well as molecular analysis based on the D1/D2 domains of the large subunit (26S) rDNA gene and the internally transcribed spacer (ITS) 1 + 2 regions. For lignocellulosic ethanol production, AFEX pretreated barley straw at $150^{\circ}C$ for 90 min was used in a simultaneous saccharification and fermentation (SSF) process using thermotolerant CHY1612. The SSF from 16% pretreated barley straw at $43^{\circ}C$ gave a saccharification ratio of 90.5%, a final ethanol concentration of 38.5 g/L, and a theoretical yield of 91.2%. These results show that K. marxianus CHY1612 has potential for lignocellulosic ethanol production through simultaneous saccharification and fermentation with further development of process.