• Korean Journal of Microbiology
• /
• v.40 no.2
• /
• pp.115-120
• /
• 2004

A Gram (-), rod-shaped bacterium in size of 1.3∼$1.8{\times}0.35{\mu}m$ inhibiting the growth of cyanobacterium (Ana-baena cylindrica) was isolated and designated NG-2 in this manuscript. This isolate showed positive reactions for catalase and oxidase, and optimal growth conditions of 35∼TEX>$40<^{\circ}C$ and pH 9.0. In a mixed-culture of A. cylindrica and the isolate, each microorganism grew inverse-proportionally, and the cyanobacterial vegetative cells almost completely disappeared within 24 hours. NG-2 lysed A. cylindrica only under light, which means that lytic activity of NG-2 was dependent on the photosynthetic activity of host. When observed under phase contrast microscope, the isolate lysed vegetative cells of A. cylindrica in scattered state in a liquid medium, whereas het-erocysts have not been lysed. When cyanobacterial cell walls have been lysed partly, NG-2 attatched around A. cylindrica filament and formed colony, then encouraged complete lysis of cyanobacterial cells. The isolate showed similar lytic activity in natural water as in an artificial medium. And lytic activity of NG-2 was enhanced when attached on expandable polystyrene bead.

• Journal of Soil and Groundwater Environment
• /
• v.15 no.1
• /
• pp.1-8
• /
• 2010

Cometabolic air sparging (CAS) is a new and innovative technology that uses air sparging principles but attempts to optimize in situ contaminant degradation by adding a growth substrate to saturated zone. CAS relies on the degradation of the primary growth substrate and cometabolic substrate transformation in the saturated zone and in the vadose zone for volatilized contaminants. In this study, we have investigated to determine MTBE degradation pattern and microbial activity variation if using propane as a primary substrate at the condition of considering air injection rate and air injection pattern. Laboratory-scale two-dimentional aquifer physical model studies were used and the experimental results were represented that the optimal conditions were as air injection rate of 1,000 mL/min and pulsed air injection pattern (15 min on/off). Over 1,000 mL/min air injection rate and continuous air injection pattern was no affected to increase DO concentration. On the other hand, Injection of propane and propane-utilizing bacteria degraded MTBE partially. And also, injection of propane- and MTBE-utilizing bacteria effectively degraded MTBE and TBA production was observed.

• Korean Journal of Veterinary Research
• /
• v.33 no.1
• /
• pp.53-60
• /
• 1993

Anaerobic bacteria are suggested to be potential source for new antibiotics. In order to search for antibiotics from domestic origin, we collected 800 soil samples across Korean locations and could isolate as many as 989 anaerobic strains. Among them 10, strains were found to have good producing capacity of antibiotics. An anaerobe was finally selected due to secreting antibiotics having high antimicrobial activity towards multiple resistant microorganism(E coli JM 83) transformed by genetic engineering technique. Its morphological, physiological and biochemical charateristics were investigated, together with antimicrobial spectrum therefrom. On antimicrobial spectrum study, substance secreted from this strain, had no activities to fungus and yeast. The selected strain showed G(+) and coccal shape, on Gram, staining and electron scanning microscopy, respectively. Biochemically this strain utilized glucose, fructose lactose, sucrose, but did not arabinose, cellulose, rhamnose, sorbitol, trehalose, mannitol. Catalase test showed negative property. Optimal growth temperature was $37^{\circ}C$. The results obtained above suggest this strain Streptococcus faecium subspp. and we named it Streptococcus sp. An-21-1.

• Microbiology and Biotechnology Letters
• /
• v.27 no.2
• /
• pp.96-101
• /
• 1999

Various side-chains are introduced to the 7-amino position of 7-aminocepha-losporanic acid (7-ACA) to make semi-synthetic cephalosporin antibiotics. In order to convert cephalosporin C (CPC) to 7-ACA, two enzymatic reactions are generally imployed. Glutary1-7-aminocephalosporanic acid (Gl-7-ACA) acylase is involved in the second step where the reaction intermediate, Gl-7-ACa is converted into 7-ACA. It was recently reported that CPC amidase can convert CPC directly into 7-ACA in a single enzymatic reaction. A study was undertaken to screen microorganisms conferring enzyme activity to convert Gl-7-ACA or CPC into 7-ACA by one or two enzymatic reactions. In order to screen the microorganisms rapidly, a non-$\beta$-lactam model compund, glutaryl-$\rho$-nitroanilide, was utilized in an early stage, thereafter the selected microorganisms were examined with real substrates. One microorganism exhibiting both Gl-7-ACA acylase and CPC amidase activities was obtained by the colorimetry method and HPLC assay, and was identified as a strain of Serratia species, designated as Serratia sp. N14.4. The optimal fermentation conditions for Serratia sp. N14.4 was pH9.0 and 3$0^{\circ}C$.

• KSBB Journal
• /
• v.13 no.4
• /
• pp.343-351
• /
• 1998

About 75 strains which utilize cholesterol as sole carbon and energy source were isolated from 10 samples of Kimchi and 18 samples of fermented fish food (2 Ojingo-jeots, salt-fermented squid ; 5 Changran-jeots, salt-fermented pollack tripe ; 5 Myungran-jeots, salt-fermented Alaska pollack roe ; 3 Gajami-sikhae-jeots, fermented flat fish ; 2 Gul-jeots, salt-fermented oyster ; a Juneo-jeots, salt-fermented shad). Among them tested, the 3T6-5Mj strain isolated from Changran-jeot showed the highest activity on cholesterol degradation. The optimal composition of medium for the producing cholesterol degradation enzyme by 3T6-5Mj strain was 1.0 g/L NH4NO3, 1.0 g/L K2HPO4, 0.1 g/L MgSO4.7H2O, 1.0 g/L FeSO4.7H2O, 1 g/L NaCl, 5 g/L Trypton, 1 g/L Cholesterol, and 5 g/L Maltose at 30$^{\circ}C$, pH 7.5, and the enzyme production reached a maximum level at 140 hours of cultivation.

• Clean Technology
• /
• v.22 no.3
• /
• pp.175-180
• /
• 2016

Tyrosinases catalyze the hydroxylation of a monophenol (monophenolase activity) and the conversion of an o-diphenol to o-quinone (diphenolase activity), which are mainly involved in the modification of tyrosine residues into 3,4-dihydroxyphenyl-alanine (DOPA) and DOPA/DOPAquinone-derived intermolecular cross-linking. Previously, we obtained a slightly acidic and cold-active tyrosinase, tyrosinase-CNK, by our recombinant protein approach. The enzyme showed optimal activity at pH 6.0 and 20 ℃ with an abnormally high monophenolase/diphenolase activity ratio and still had approximately 50% activity compared with the highest activity even in ice water. Here, we investigated reaction stability of the recombinant tyrosinase-CNK as a psychrophilic enzyme. The enzyme showed remarkable thermal stability at 0 ℃ and the activity was well conserved in repeated freeze-thaw cycles. Although water-miscible organic solvent as reaction media caused the activity decrease of tyrosinase-CNK as expected, the enzyme activity was not additionally decreased with increased concentration in organic solvents such as ethanol and acetonitrile. Also, the enzyme showed high salt tolerance in chaotropic salts. It was remarkably considered that 2⁺ metal ions might inhibit the incorporation of Cu²⁺ into the active site. We expect that these results could be used to design tyrosinase-mediated enzymatic reaction at low temperature for the production of catechols through minimizing unwanted self-oxidation and enzyme inactivation.

• Asian-Australasian Journal of Animal Sciences
• /
• v.31 no.4
• /
• pp.607-615
• /
• 2018

Objective: This study was conducted to isolate the cellulolytic microorganism from the rumen of Holstein steers and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods: To isolate anaerobic microbes having endoglucanase, rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 98% similarity with Eubacterium cellulosolvens (E. cellulosolvens) Ce2 (Accession number: AB163733). The Cel5A from isolated E. cellulolsovens sp. was cloned using the published genome sequence and expressed through the Escherichia coli BL21. Results: The maximum activity of recombinant Cel5A (rCel5A) was observed at $50^{\circ}C$ and pH 4.0. The enzyme was constant at the temperature range of $20^{\circ}C$ to $40^{\circ}C$ but also, at the pH range of 3 to 9. The metal ions including $Ca^{2+}$, $K^+$, $Ni^{2+}$,$Mg^{2+}$, and $Fe^{2+}$ increased the endoglucanase activity but the addition of $Mn^{2+}$, $Cu^{2+}$, and $Zn^{2+}$ decreased. The Km and Vmax value of rCel5A were 14.05 mg/mL and $45.66{\mu}mol/min/mg$. Turnover number, Kcat and catalytic efficiency, Kcat/Km values of rCel5A was $96.69(s^{-1})$ and 6.88 (mL/mg/s), respectively. Conclusion: Our results indicated that rCel5A of E. cellulosolvens isolated from Holstein steers had a broad pH range with high stability under various conditions, which might be one of the beneficial characteristics of this enzyme for possible industrial application.

• Applied Biological Chemistry
• /
• v.36 no.6
• /
• pp.539-546
• /
• 1993

This paper describes the purification and partial characteristics of aminopeptidase from Microccus sp. LL3 to utilize the microorganism as a potential agent for industrial application for the purpose of shortening ripening period of cheddar cheese. The optimal temperature and pH for enzyme activity were $35^{\circ}C$ and 7.0, respectively for L-leucine-p-nitroanilide as substrate. The enzyme remained stable for 10 minutes up to $50^{\circ}C$. The activity of aminopeptidase was stimulated by $Mg^{++}$ ion but strongly inhibited by $Hg^{++}$, metal complexing reagents, ethylenediaminetetraacetate (EDTA) and 1,10-phenanthroline. The enzyme was thought to be metallopeptidase. This enzyme had a broad substrate specificity, but was inactive on peptide with arginine as N-terminal amino acid. An intracellular aminopeptidase from Micrococcu sp. LL3 was purified by chromatography on DEAE-Sephacel and filtration on Sepacryl S-300. The enzyme has a molecular weight of 43,500.

• Journal of Microbiology and Biotechnology
• /
• v.12 no.6
• /
• pp.921-928
• /
• 2002

Microorganism producing extracellular CFTase was isolated from soil and designated as Bacillus polymyxa MGL21. The gene encoding the CFTase (cft) from B. polymyxa MGL21 was cloned and sequenced. The ORF of the cf gene was composed of 3,999 nucleotides, encoding a protein (1,333 amino acids) with a predicted molecular mass of 149,375 Da. Sequence analysis indicated that CFTase was divided into five distinct regions. CFTase contained three regions of repeat sequences at the N-terminus and C-terminus. The endo-inulinase region of homology (ERH) of CFTase was similar to that of Pseudomonas mucidolens endo-inulinase ($50\%$ identity, 259 amino acids). Furthermore, CFTase possessed a highly conserved core region, which is considered to be functional for the hydrolysis reaction of inulin. The cft gene was expressed in a His-tagged form in Escherichia coli cells, and the His-tagged CFTase was purified to homogeneity. The optimal temperature and pH for CFTase activity were found to be $50^{\circ}C$ and 9.0, respectively. The enzyme activity was completely inhibited by 10 mM $Ag^+\;and\;Cu^2+$. Thin-layer chromatography analyses indicated that CFTase catalyzed not only the cyclization reaction ut also disproportionation and hydrolysis reactions as well.

• Environmental Engineering Research
• /
• v.19 no.4
• /
• pp.309-316
• /
• 2014

The water quality improvement of golf course ponds, as representative stagnant stream channels, was evaluated by applying a biological activated carbon (BAC) process composed of four consecutive activated carbon reactors. The study was performed from autumn to winter in order to evaluate the feasibility of the BAC process under low temperature conditions. In the study, water quality of pond A (target pond) and pond B (reference pond) were monitored. Pond water was pumped into the BAC process, and was then returned to the pond after treatment. The optimal conditions were determined to be 2 hr of empty bed contact time (EBCT) at a temperature above $4^{\circ}C$, in which improvements of chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP) of pond A compared to pond B were 3.62%, 3.48% and 1.81%, respectively. On the other hand, as the temperature was below $4^{\circ}C$, some degree of water quality improvement was achieved even when EBCT were 1 or 0.5 hr, suggesting that the BAC process can be successfully applied for the improvement of pond water quality in winter months. The values of biomass concentration and microorganism activity in each condition were highest where 2 hr of EBCT was applied at a temperature above $4^{\circ}C$, but values were similar throughout all treatment conditions, and thus, adsorption is considered to be the dominant factor affecting process efficiency. From the denaturing gel gradient electrophoresis (DGGE) results, no significant differences were observed among the activated carbon reactors, suggesting that the number of reactors in the system could be decreased for a more compact application of the system.