• Journal of Microbiology and Biotechnology
• /
• v.11 no.5
• /
• pp.863-869
• /
• 2001

A systematic study of microbial fuel cells comprised of alkalophilic Bacillus sp. B-31 has been carried out under various operating conditions. A significant amount of electricity was generated when redox mediators were used. Among the phenothiazine-type redox dyes tested, azure A was found to be the most effective both in maintaining a high cell voltage and for the long-term operation. The maximum efficiency was and for the long-term operation. The maximum efficiency was obtained at ca. $50^{\circ}C$ giving an open circuit voltage of 0.7V. A small change in temperature did not significantly affect the cell performance, but a rapid decrease in performance was observed below $20^{\circ}C$ and above $70^{\circ}C$. It was noticeable that fuel cell efficiency and discharge pattern depended strongly on the carbon source used in the initial culture medium. Regardless of the initial carbon sources, only glucose and trehalose were utilized as substrates. Galactose, however, was not substantially utilized except when galactose was used in the initial medium. Glucose, in particular, showed $87\%$ coulombic efficiency, which was the highest value ever reported, when Bacillus sp. was cultured in a maltose-containing medium. This study demonstrates that highly efficient microbial fuel cells can be constructed with alkalophilic microorganisms by fine-tuning the operating conditions and by carefully selecting carbon sources in the initial culture medium.

• Journal of Microbiology and Biotechnology
• /
• v.8 no.5
• /
• pp.501-508
• /
• 1998

An alkalophilic mi.croorganism (strain YB380), which produces yeast cell wall hydrolase extracellulary, was isolated from Korean soil. The rod-shaped cells were 0.3~0.4 by 2~4${\mu}{\textrm}{m}$ long, motile, aerobic, gram-positive, and spore-forming. The color of the colony was light yellow. The temperature range for growth at pH 9.0 was 25 to $45{\circ}C, with optimum growth at $35{\circ}C. The pH range for growth at $35{\circ}C was 8 to 11 with an optimum pH of 9.0. Therefore, the strain YB380 is an obligate alkalophile. The 16S rRNA of strain YB380 has a 99% sequence similarity with that of Bacillus alcalophilus. On the basis of physiological properties, cell wall fatty acid composition, and phylogenetic analysis, we propose that the isolated strain is Bacillus alcalophilus. The yeast cell wall hydrolase from Bacillus alcalophilus subsp. YB380 has been purified and partially characterized. The molecular weight was estimated to be 27,000 daltons with an optimum temperature and pH of $60{\circ}C and 9.0, respectively. The N-terminal amino acid sequence of the enzyme was analyzed as Gln- Thr- Val- Pro- Trp- Gly- Ile- Asn- Arg- Val.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.5
• /
• pp.667-667
• /
• 2000

Pseudomonas sp. BK7, an alkalophile, displayed the highest growth and protease activity when grown in a fermenter which was controlled at a pH level of 9.0, and the enzyme production was significantly enganced by the increase of agitation speed. Two formas of alkaline proteases (BK7-1 and BK7-2) were fractionated and purified to near homogeneity. Protease BK7-1 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B, DEAE-Sepharose, and Sephadex G-75 column chromatographies. The molecular weights of proteases BK7-1 and BK7-2 determined by gel filtration chromatography were 20,700 and 40,800, respectively. The $K_m$ value, isoelectric point, and optimum pH of protease BK7-1 were 2.55 mg/ml, 11.0 and 11.0, respectively, whereas those of protease BK7-2 were 1.57 mg/ml, 7.2, and 10.0, respectively. Both protease were practically stable in the pH range of 5-11. The optimum temperatures for the activities of both protease BK7-1 and BK7-2 were 50℃ and 45℃, respectively. About 56% of the original protease BK7-2 activity remained after being treated at 50℃ for 30 min but protease BK7-1 was rapidly inactivated at above 25℃. Both proteases were completely inhibited by phenylmethane sulfonyl fluoride, a serine protease inhibitor. Protease BK7-2 was stable against EDTA, EGTA, STP, and detergents such as SDS and LAS, whereas protease BK7-1 was found to be unstable.

• Microbiology and Biotechnology Letters
• /
• v.35 no.4
• /
• pp.298-303
• /
• 2007

A halophilic bacterium, H. subglaciescola DH-1, grew at 2.0 M salinity, but did not grow at 0.8 M salinity when cultivated at higher temperature ($40^{\circ}C$) than optimum ($30^{\circ}C$). When the cell extract of strain DH-1 was heated at $50^{\circ}C$ for 60 min in the absence of NaCl, isocitrate dehydrogenase and malate dehydrogenase lost their activities, but when it was heated in the presence of 2.0 M NaCl, the activity was maintained. Meanwhile, the cell extract of E. coli did not catalyze the reduction of $NAD^+$ to NADH coupled with the oxidation of isocitrate and malate at higher salinities than 1.0 M. The pH range for DH-1 was 7 to 10, and that for E. coli was 5 to 9. DH-1 was not grown in conditions with sodium salts other than NaCl.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.5
• /
• pp.677-684
• /
• 2000

Pseudomonas sp. BK7, an alkalophile, displayed the highest growth and protease activity when grown in a fermenter which was controlled at a pH level of 9.0, and the enzyme production was significantly enhanced by the increase of agitation speed. Two forms of alkaline proteases (BK7-1 and BK7-2) were fractionated and purified to near homogeneity. Protease BK7-1 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B, DEAE-Sepharose, and Sephadex G-75 column chromatographies. The molecular weights of proteases BK7-1 and BK7-2 determined by gel filtration chromatography were 20,700 and 40,800, respectively. The $K_m$ value, isoelectric point, and optimum pH of protease BK7-1 were 2.55 mg/ml, 11.0, and 11.0, respectively, whereas those of protease BK7-2 were 1.57 mg/ml, 7.2, and 10.0, respectively. Both proteases were practically stable in the pH range of 5-11. The optimum temperatures for the activities of both protease BK7-1 and BK7-2 were $50^{\circ}C$ and $45^{\circ}C$, respectively. About 56% of the original protease BK7-2 activity remained after being treated at $50^{\circ}C$ for 30 min but protease BK7-1 was rapidly inactivated at above $25^{\circ}C$. Both proteases were completely inhibited by phenylmethane sulfonyl fluoride, a serine protease inhibitor. Protease BK7-2 was stable against EDTA, EGTA, STP, and detergents such as SDS and LAS, whereas protease BK7-1 was found to be unstable.