• 한국미생물·생명공학회지
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• 제43권3호
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• pp.236-240
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• 2015

초고온성 고세균 Thermococcus onnurineus NA1은 개미산, 일산화탄소, 또는 전분 등을 이용해서 수소를 생산하는 것으로 알려져 있다. 본 연구에서는 T. onnurineus NA1의 고정화 세포를 이용한 수소생산을 고찰하였다. 고정화 실험결과, T. onnurineus NA1은 표면에 아민기가 코팅된 규조토 담체에 정전기적 인력에 의해 효과적으로 고정화되었고, 1 g의 담체에 고정화 될 수 있는 최대 세포의 양은 71.7 mg-dcw로 확인되었다. 고정화 세포를 이용한 세 번의 반복회분식 배양을 통해 개미산으로부터 수소생산 특성을 고찰하였고, 그 결과 배양이 반복됨에 따라 고정화 세포 농도의 증가에 기인하여 초기수소생산속도가 2.3 에서 4.0 mmol l⁻¹ h⁻¹로 상당량 증가됨이 관찰되었다. 따라서, T. onnurineus NA1의 고정화세포 시스템은 수소생산을 위한 좋은 대안이 될 수 있을 것으로 사료된다. 본 연구는 초고온성 고세균의 고정화세포를 수소생산에 적용한 첫 번째 사례이다.

• 한국수소및신에너지학회논문집
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• 제22권5호
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• pp.671-677
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• 2011

A hyperthermophilic archaeon, $Thermococcus$ $onnurineus$ NA1 was studied to investigate its fermentation characteristics using various carbon sources including formate, maltose and carbon monoxide during the anaerobic batch cultivation at $80^{\circ}C$. Formate was the best carbon source for cell growth and hydrogen production among others. In the batch culture on formate, it was found that the cell concentration increased exponentially by 12 hrs of culture, after which the cell growth and formate consumption was retarded. Hydrogen production was continued more than 24 hrs although the cell growth was ceased at 18 hrs. Hydrogen production rate was directly correlated with the cell growth and formate degradation up to 18 hrs, and the average hydrogen production yield was 1.05 mole-$H_2$/mole-formate. Cell growth and hydrogen production were optimized at the initial pH 6-7, while inhibited at the initial pH lower than 5 and higher than 9.

• Ocean and Polar Research
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• 제39권4호
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• pp.269-277
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• 2017

Medium compositions for the hyperthermophilic archaeon, Thermococcus onnurineus NA1 was statistically optimized to enhance formate-driven hydrogen ($H_2$) production by using response surface methodology. From the Plackett-Burman design-based experiment, it was confirmed that among the minor components of medium such as KCl, $MgSO_4$, $NH_4Cl$, Cystein-HCl, trace elements, Fe-EDTA and $CaCl_2$, the trace elements were screened as the only positively effective components with respect to $H_2$ production. Subsequently, the optimal concentrations of the trace elements and the major components of a medium such as NaCl, yeast extract and sodium formate were determined from the five-level central composite design (CCD)-based experiment. The resulting quadratic model predicted the maximum $H_2$ production of 46.6 mmol/L in serum bottle and it was validated experimentally using the optimal medium initially supplemented with 26.70 g/L of NaCl, 9.81 g/L of sodium formate, 3.50 g/L of yeast extract and 4.59 mL/L of trace elements. From the duplicate batch cultivations in the fermentor using the optimized medium, the a maximum $H_2$ production rate up to 71.8 mmol/L/h could be obtained, which was a 65% enhanced value compared with that obtained using the control medium, showing the high efficiency of the optimized medium.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1826-1831
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• 2006

A novel hyperthermophilic, anaerobic, heterotrophic archaeon, designated strain $NA1^T$, was isolated from a deep-sea hydrothermal vent area (depth, 1,650 m) within the Papua New Guinea-Australia-Canada-Manus (PACMANUS) field. Cells of this strain were motile by means of polar flagella, coccoid-shaped with a diameter of approximately $0.5-1.0{\mu}m$, and occurred as single cells. Optimal temperature, pH, and NaCl concentration for growth were $80^{\circ}C$, 8.5, and 3.5%, respectively. The new isolate was an obligate heterotroph that utilized yeast extract, beef extract, tryptone, peptone, casein, and starch as carbon and energy sources. Elemental sulfur was required for growth and was reduced to hydrogen sulfide. The G+C content of the genomic DNA was 52.0 mol%. Phylogenetic analysis of the 16S rRNA gene indicated that strain $NA1^T$ belongs to the genus Thermococcus, and the organism is most closely related to T. gorgonarius, T. peptonophilus, and T. celer; however, no significant homology was observed among species by DNA-DNA hybridization. Strain $NA1^T$ therefore represents a novel species for which the name Thermococcus onnurineus sp. novo is proposed. The type strain is $NA1^T$ (=KCTC 10859, =JCM 13517).

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2009년도 International Meeting of the Microbiological Society of Korea
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• pp.92-93
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• 2009

• Journal of Microbiology and Biotechnology
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• 제17권8호
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• pp.1242-1248
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• 2007

Genomic analysis of a hyperthermophilic archaeon, Thermococcus onnurineus NA1 [1], revealed the presence of an open reading frame consisting of 1,377 bp similar to ${\alpha}$-amylases from Thermococcales, encoding a 458-residue polypeptide containing a putative 25-residue signal peptide. The mature form of the ${\alpha}$-amylase was cloned and the recombinant enzyme was characterized. The optimum activity of the enzyme occurred at $80^{\circ}C$ and pH 5.5. The enzyme showed a liquefying activity, hydrolyzing maltooligosaccharides, amylopectin, and starch to produce mainly maltose (G2) to maltoheptaose (G7), but not pullulan and cyclodextrin. Surprisingly, the enzyme was not highly thermostable, with half-life ($t_{1/2}$) values of 10 min at $90^{\circ}C$, despite the high similarity to ${\alpha}$-amylases from Pyrococcus. Factors affecting the thermostability were considered to enhance the thermo stability. The presence of $Ca^{2+}$ seemed to be critical, significantly changing $t_{1/2}$ at $90^{\circ}C$ to 153 min by the addition of 0.5 mM $Ca^{2+}$. On the other hand, the thermostability was not enhanced by the addition of $Zn^{2+}$ or other divalent metals, irrespective of the concentration. The mutagenetic study showed that the recovery of zinc-binding residues (His175 and Cys189) enhanced the thermo stability, indicating that the residues involved in metal binding is very critical for the thermostability.

• Ocean and Polar Research
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• 제38권3호
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• pp.225-234
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• 2016

This project sought to conduct an economic feasibility study regarding the commercial production of bio-hydrogen by the marine hyperthermophilic archaeon, Thermococcus onnurineus NA1 using carbon monoxide-containing industrial off-gas. We carried out the economic evaluation of the bio-hydrogen production process using the raw material of steel mill by-product gas. The process parameter was as follows: $H_2$ production rate was 5.6 L/L/h; the conversion of carbon monoxide was 60.7%. This project established an evaluation criterion for about 10,000 tonne/year. Inflation factors were considered as 3%. The operating costs were recalculated based on prices in 2014. The total investment required for development was covered 30% by capital and 70% by a loan. The operation cost for the 0.5-year test and integration, and the cost for the first three months in the 50% production period were considered as the working capital in the cost estimation. The costs required for the rental of office space, facilities, and other related costs from the construction through to full-scale production periods were considered as continuing expenses. Materials, energy, waste disposal and other charges were considered as the operating cost of the development system. Depreciation, tax, maintenance and repair, insurance, labor, interest rate charges, general and administrative costs, lubrication and miscellaneous expenses were also calculated. The hydrogen price was set at US$ 4.15/kg for the economic evaluation. As a result, the process was considered to be economical with the payback period of 6.3 years, NPV of 18 billion Won and IRR of 26.7%.

• 한국수소및신에너지학회논문집
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• 제29권4호
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• pp.307-316
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• 2018

In this study, the failure mode and effect analysis (FMEA) of hydrogen production process by using the Thermococcus onnurineus NA1 was conducted and advanced methodology to compensate the weakness of previous FMEA methodology was applied. To bring out more quantitative and precise FMEA result for bio-hydrogen production process, fuzzy logic and potential loss cost estimated from ASPEN Capital Cost Estimator (ACCE) was introduced. Consequently, risk for releasing the flammable gases via internal leakage of steam tube which to control the operating temperature of main reactor was caution status in FMEA result without applying the fuzzification and ACCE. Moreover, probability of the steam tube plugging caused by solid property like medium was still caution status. As to apply the fuzzy logic and potential loss cost estimated from ACCE, a couple of caution status was unexpectedly upgraded to high dangerous status since the potential loss cost of steam tube for main reactor and decrease in product gases are higher than expected.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2008년도 International Meeting of the Microbiological Society of Korea
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• pp.119-120
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• 2008