• Journal of Microbiology and Biotechnology
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• 제30권6호
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• pp.930-936
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• 2020

The red seaweed Gracilaria verrucosa has been used for the production of bioethanol. Pretreatment for monosaccharide production was carried out with 12% (w/v) G. verrucosa slurry and 500 mM HNO₃ at 121℃ for 90 min. Enzymatic hydrolysis was performed with a mixture of commercial enzymes (Cellic C-Tec 2 and Celluclast 1.5 L; 16 U/ml) at 50℃ and 150 rpm for 48 h. G. verrucosa was composed of 66.9% carbohydrates. In this study, 61.0 g/L monosaccharides were obtained from 120.0 g dw/l G. verrucosa. The fermentation inhibitors such as hydroxymethylfurfural (HMF), levulinic acid, and formic acid were produced during pretreatment. Activated carbon was used to remove HMF. Wild-type and adaptively evolved Saccharomyces cerevisiae, Candida lusitaniae, and Kluyveromyces marxianus were used for fermentation to evaluate ethanol production.

• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.318-324
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• 1998

A ${\beta}$-galactosidase with high transgalactosylic activity was purified from a Bacillus species, registered as KFCC10855. The enzyme preparation showed a single protein band corresponding to a molecular mass of 150 kDa on SDS-PAGE and gave a single peak with the estimated molecular mass of 250 kDa on Sephacryl S-300 gel filtration, suggesting that the enzyme is a homodimeric protein. The amino acid and sugar analyses revealed that the enzyme is a glycoprotein, containing 19.2 weight percent of sugar moieties, and is much more abundant in hydrophilic amino acid residues than in hydrophobic residues, the mole ratio being about 2:1. The pI and optimum pH were determined to be 5.0 and 6.0, respectively. Having a temperature optimum at $70^{\circ}C$ for the hydrolysis of lactose, the enzyme showed good thermal stability. The activity of the enzyme preparation was markedly increased by the presence of exogenous Mg (II) and was decreased by the addition of EDTA. Among the metal ions examined, the most severely inhibitory effect was seen with Ag (I) and Hg (II). Further, results of protein modification by various chemical reagents implied that 1 cysteine, 1 histidine, and 2 methionine residues occur in certain critical sites of the enzyme, most likely including the active site. Enzyme kinetic parameters, measured for both hydrolysis and transgalactosylation of lactose, indicated that the enzyme has an excellent catalytic efficiency for formation of the transgalactosylic products in reaction mixtures containing high concentrations of the substrate.

• Journal of Microbiology and Biotechnology
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• 제26권7호
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• pp.1259-1266
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• 2016

Bioethanol was produced from Kappaphycus alvarezii seaweed biomass using separate hydrolysis and fermentation (SHF). Pretreatment was evaluated for 60 min at 121℃ using 12% (w/v) biomass slurry with 364 mM H₂SO₄. Enzymatic saccharification was then carried out at 45℃ for 48 h using Celluclast 1.5 L. Ethanol fermentation with 12% (w/v) K. alvarezii hydrolyzate was performed using the yeasts Saccharomyces cerevisiae KCTC1126, Kluyveromyces marxianus KCTC7150, and Candida lusitaniae ATCC42720 with or without prior adaptation to high concentrations of galactose. When non-adapted S. cerevisiae, K. marxianus, and C. lusitaniae were used, 11.5 g/l, 6.7 g/l, and 6.0 g/l of ethanol were produced, respectively. When adapted S. cerevisiae, K. marxianus, and C. lusitaniae were used, 15.8 g/l, 11.6 g/l, and 13.4 g/l of ethanol were obtained, respectively. The highest ethanol concentration was 15.8 g/l, with Y_EtOH = 0.43 and Y_T% = 84.3%, which was obtained using adapted S. cerevisiae.

• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.332-339
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• 2010

The lipase from Mucor racemosus NRRL 3631 was partially purified by fractional precipitation using 60% ammonium sulfate, which resulted in a 8.33-fold purification. The partially purified lipase was then immobilized using different immobilization techniques: physical adsorption, ionic binding, and entrapment. Entrapment in a 4% agar proved to be the most suitable technique (82% yield), as the immobilized lipase was more stable at acidic and alkaline pHs than the free enzyme, plus 100% of the original activity was retained owing to the thermal stability of the immobilized enzyme after heat treatment for 60 min at $45^{\circ}C$. The calculated half-lives (472.5, 433.12, and 268.5 min at 50, 55, and $60^{\circ}C$, respectively) and the activation energy (9.85 kcal/mol) for the immobilized enzyme were higher than those for the free enzyme. Under the selected conditions, the immobilized enzyme had a higher $K_m$ (11.11 mM) and lower $V_{max}$ (105.26 U/mg protein) when compared with the free enzyme (8.33 mM and 125.0 U/mg protein, respectively). The operational stability of the biocatalyst was tested for both the hydrolysis of triglycerides and esterification of fatty acids with glycerol. After 4 cycles, the immobilized lipase retained approximately 50% and 80% of its original activity in the hydrolysis and esterification reactions, respectively.

• KSBB Journal
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• 제28권5호
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• pp.282-286
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• 2013

The seaweed, Gelidium amansii, was fermented to produce bioethanol. Optimal pretreatment condition was determined as 94 mM $H_2SO_4$ and 8% (w/v) seaweed slurry at $121^{\circ}C$ for 60 min. The mono sugars of 40.4 g/L with 67% of conversion from total carbohydrate of 60.6 g/L with 80 g dw/L G. amansii slurry were obtained by thermal acid hydrolysis pretreatment and enzymatic saccharification. G. amansii hydrolysate was used as the substrate for ethanol production by Kluyveromyces marxianus KCTC 7150 and Candida tropicalis KCTC 7212 using 5L fermentor. The ethanol productions by K. marxianus KCTC 7150 and C. tropicalis KCTC 7212 were 17.8 g/L with $Y_{EtOH}$ of 0.48 at 120 h and 19.3 g/L with $Y_{EtOH}$ of 0.50 at 120 h, respectively.

• 상하수도학회지
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• 제29권2호
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• pp.183-192
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• 2015

Response surface methodology (RSM) based on a Box-Behnken Design (BBD) was applied to optimize the thermal-alkaline pre-treatment operating conditions for anaerobic digestion of flotation scum in food waste leachate. Three independent variables such as thermal temperature, NaOH concentration and reaction time were evaluated. The maximum methane production of 369.2 mL $CH_4/g$ VS was estimated under the optimum conditions at $62.0^{\circ}C$, 10.1% NaOH and 35.4 min reaction time. A confirmation test of the predicted optimum conditions verified the validity of the BBD with RSM. The analysis of variance indicated that methane production was more sensitive to both NaOH concentration and thermal temperature than reaction time. Thermal-alkaline pretreatment enhanced the improvement of 40% in methane production compared to the control experiment due to the effective hydrolysis and/or solubilization of organic matters. The fractions with molecular weight cut-off of scum in food waste leachate were conducted before and after pre-treatment to estimate the behaviors of organic matters. The experiment results found that thermal-alkaline pre-treatment could reduce the organic matters more than 10kD with increase the organic matters less than 1kD.

• 신재생에너지
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• 제10권1호
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• pp.20-29
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• 2014

The present study investigated the effects of thermal pre-treatment on the enhancement of anaerobic biodegradability of waste activated sludge at varied TS concentration levels. The activated sludges were thermally oxidized for 30 minutes at $80{\sim}200^{\circ}C$ with varied TS concentrations (2%, 4% and 6%). and then, sludge characteristics, solubilization efficiency and methane production yield of thermally pre-treated sludges were analyzed. The higher the temperature in the thermal pre-treatment, the higher the concentration levels of dissolved matters such as $SCOD_{Cr}$, $NH_4{^+}$ and VFAs, which indicates that the thermal pre-treatment facilitates the hydrolysis and acid fermentation. Furthermore, the solubilization efficiency was increased in proportion to the temperature rise at all TS concentrations and was reached at 68.9%, 55.6% and 53.1%, respectively, at $200^{\circ}C$. In the BMP test of the pre-treated sludges, higher methane production yields were observed as 0.313. 0.314 and $0.299m^3\;CH_4/kg\;VS_{add}$ at the condition of TS 2% ($160^{\circ}C$), 4% ($160^{\circ}C$) and 6% ($180^{\circ}C$), respectively, and degradation rate was increased by 84%, 79% and 65% compared with non-pretreated waste activated sludge. These findings suggest the effectiveness of thermal pre-treatment of waste activated sludge for anaerobic biodegradable process.

• 한국환경과학회지
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• 제31권7호
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• pp.609-616
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• 2022

In this study, the effect of different reaction times for thermal-alkaline pretreatment on the solubilization and biogasification of polyhydroxybutyrate (PHB) were evaluated. Thermal-alkaline pretreatment tests were performed at 73 ℃ and pH 13 at 0-120 h reaction times. The mesophilic anaerobic batch tests were performed with untreated and pretreated PHB samples. The increase in the pretreatment reaction time results in a 52.8-98.8% increase of the abiotic solubilization efficiency of the PHB samples. The reaction time required to achieve solubilization efficiencies of 50%, 90%, and 95% were 10.5, 52.0, and 89.6 h, respectively. The biogasification of the untreated PHB samples achieved a specific methane production rate of 3.6 mL CH₄/g VSS/d and require 101.3 d for complete biogasification. The thermal-alkaline pretreatment significantly improved specific methane production rate (10.2-16.0 time increase), lag time (shortened by 76-81%), and time for complete biogasification (shortened by 21-83%) for the biogasification of the PHB samples when compared to those of the untreated PHB samples. The improvement was higher as the reaction time of the thermal-alkaline pretreatment increased. The findings of this study could be used as a valuable reference for the optimization of the biogasification process in the treatment of PHB wastes.

• Journal of Microbiology and Biotechnology
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• 제23권6호
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• pp.818-825
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• 2013

We isolated and functionally characterized the ${\alpha}$- and ${\beta}$-subunits (ApCpnA and ApCpnB) of a chaperonin from Aeropyrum pernix K1. The constructed vectors pET3d-ApCpnA and pET21a-ApCpnB were transformed into E. coli Rosetta (DE3), BL21 (DE3), or CodonPlus (DE3) cells. The expression of ApCpnA (60.7 kDa) and ApCpnB (61.2 kDa) was confirmed by SDS-PAGE analysis. Recombinant ApCpnA and ApCpnB were purified by heat-shock treatment and anion-exchange chromatography. ApCpnA and ApCpnB were able to hydrolyze not only ATP, but also CTP, GTP, and UTP, albeit with different efficacies. Purified ApCpnA and ApCpnB showed the highest ATPase, CTPase, UTPase, and GTPase activities at $80^{\circ}C$. Furthermore, the addition of ApCpnA and ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively. In particular, the addition of ATP or CTP to ApCpnA and ApCpnB resulted in the most effective prevention of thermal aggregation and inactivation of CS and ADH. The ATPase activity of the two chaperonin subunits was dependent on the salt concentration. Among the ions we examined, potassium ions were the most effective at enhancing the ATP hydrolysis activity of ApCpnA and ApCpnB.

• 한국미생물·생명공학회지
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• 제45권3호
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• pp.236-242
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• 2017

본 연구에서는 미역을 이용하여 초고온 열산 가수분해, 효소 당화, 발효과정을 거쳐 아세톤, 부탄올, 에탄올을 생성하는 실험에 대해 진행하였다. 초고온 열산 가수분해에서의 최적 조건은 10%의 slurry, 270 mM의 황산, $160^{\circ}C$에서의 7.5분이었다. 초고온 열산 가수분해는 열처리 시간을 줄이고 적은 농도의 황산을 사용해도 더 많은 당과 적은 저해물질을 생성해 낸다는 장점이 있다. 효소 당화에서는 Viscozyme L (${\beta}-glucanase$, Novozymes)을 12 unit/ml으로 처리하는 것이 25.1 g/l로 가장 많은 단당을 생성했다. 발효에서는 C. acetobutylicum KCTC 1724이 비교적 낮은 pH 5.0에서 많은 아세톤, 부탄올, 에탄올을 생성하는 장점이 있었지만 mannitol을 모두 소비하지 못하는 단점이 있어 고농도의 mannitol 배지에 순치한 C. acetobutylicum KCTC 1724을 사용하여 발효를 진행하였다. 그 결과, 아세톤, 부탄올, 에탄올이 각각 0.99 g/l, 5.62 g/l, 2.44 g/l로 순치하지 않은 C. acetobutylicum KCTC 1724를 이용해 발효했을 때 보다 부탄올은 2.45 g/l, 에탄올은 1.10 g/l 증가했으며 수율($Y_{ABE}$)은 0.24에서 0.37로 증가했다.