• Microbiology and Biotechnology Letters
• /
• v.44 no.2
• /
• pp.130-139
• /
• 2016

A bacterial strain, producing an excellent lipolytic enzyme, was isolated from the intestinal tracts of an earthworm (Eisenia fetida). The strain was identified as Aeromonas hydrophila by phenotypic, chemotaxonomic characteristics and 16S ribosomal DNA analysis, and was designated as Aeromona hydrophila PL43. The lipolytic enzyme from A. hydrophila PL43 was purified via 35−45% ammonium sulfate precipitation, DEAE-sepharose fast flow ion-exchange, and sephacryl S-300HR gel filtration chromatography. The yield of the purified enzyme was 3.7% and 2.5% of the total activity of crude extracts with p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as substrates, respectively. The molecular weight of the purified enzyme was approximately 74 kDa using gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and zymography. The optimal activity of purified enzyme was observed at 50℃ and pH 8.0 using pNPB, and 60℃ and pH 8.0 using pNPP. The purified enzyme was stable in the ranges 20− 60℃ and pH 7.0−10.0. The activity of purified enzyme was inhibited by PMSF, pepstatin A, Co²⁺, Cu²⁺, and Fe²⁺, but was recovered by metal chelating of EDTA. The K_m and V_max values of the purified enzyme were 1.07 mM and 7.27 mM/min using pNPB and 1.43 mM and 2.72 mM/min using pNPP, respectively.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1997.04a
• /
• pp.84-84
• /
• 1997

장내세균은 음식물, 스트레스, 생활환경에 의해 영향을 받으며 그 결과 장내 세균이 생산하는 효소활성도 영향을 받는다. 장내미생물효소는 질병과 밀접한 관계를 갖고 있으며 장내의 pH와 효소저해제에 의해 영향을 받는 데 장내의 높은 pH에 의해 $\beta$-glucosidase, $\beta$-glucuronidase, tryptophanase 등의 장내유해효소활성이 유도되므로 장내의 pH를 낮춤으로써 효소활성을 저하시킬 수 있다. Bifidobacterium은 장내에서 lactic acid, acetic acid를 생산하여 장내의 pH를 낮추며 유해균의 증식을 억제하고 유해효소의 활성을 억제하는 역할을 할 것으로 기대된다. 특히 한국인으로부터 분리된 유산균일 경우 한국인의 장내에 가장 잘 정착되며 장내미생물의 유해효소를 효과적으로 억제할 것으로 생각된다.

• Korean Journal of Microbiology
• /
• v.25 no.3
• /
• pp.189-197
• /
• 1987

The aim of the present study was to investigate the effect of glutamate on the biosynthesis of tylosin. Activities of enzymes involved in the metabolic pathway of glutamate to form tylactone, an essential precursor of tylosin, were determined using Streptomyces fradiae grown at different concentration of glutamate. As results, it was found that cell growth and tylactone formation was controlled by the metabolic flux of oxaloacetate. It was clear that cell growth was favored by the activities of citrate synthase and aspartate aminotransferase, while the tylactone synthesis was stimulated by the activity of methylmalonyl-CoA carboxyltransferase. Therefore it was concluded that channelling of oxaloacetate was a point for favoring either cell growth or tylosin biosynthesis.

• Microbiology and Biotechnology Letters
• /
• v.8 no.4
• /
• pp.229-235
• /
• 1980

Pectin transeliminase (PTE) was produced by Saccharomyces cerevisiae, Schizosaccharomyces pombe 4683, and Aspergillus niger in the media containing 2% pectin and examined for its characteristics. The Production of the enzyme was higher by Asp. niger than by the two yeast strains, showing that the PTE activity was proportional to reducing power. The enzyme was proved to reduce pectin and produce 4, 5- unsaturated galacturonic acid. The optimum activity of the PTE was found to be at pH 6.0 and $50^{\circ}C$. The activities of these enzyme were stable below $50^{\circ}C$ but decreased at the higher temperature. Substrate inhibition of the PTE activities was appeared at high concentrations of pectin. Those PTE activities were increased under 0.6M of KCI and NaCI, but that maximal activities at the concentration of 0.2M MgC $l_2$.

• Korean Journal of Microbiology
• /
• v.23 no.3
• /
• pp.177-183
• /
• 1985

Some properties of an extracellular cytosine deaminase produced from Arthrobacter sp.JH-13 were examined after 20-80% of ammonium sulfate fractionation. Among some substrates, this enzyme utilized cytosine and 5-fluorocytosine as a substrate. The optimum pH and temperature for the activity of this enzyme were found to be near 8.0 and $40^{\circ}C$, respectively. The ensyme was more stable in 0.2M of Tris-HCl buffer than 0.2M of potassium phosphate buffer. The enzyme was generally stable below $50^{\circ}C$, but inactivated completely at $70^{\circ}C$. 1mM of $Fe^{3+},\;K^+\;and\;Na^+$ increased the enzyme activity, but 0.01mM of $Co^{2+},\;Cu^{2+},\;Ni^{2+},\;Hg^{2+},\;Ag^{2+},\;Zn^{2+},\;Ba^{2+},\;and\;Mg^{2+}$ markedly inactivated the enzyme activity. 0.1mM of p-chloromercuribenzoate, trichloroacetic acid, and N-ethylmaleimide compleyely inhibited the enzyme activity, but 0.1mM of 2-mercaptoethanol slightly increased the enzyme activity.

• Korean Journal of Microbiology
• /
• v.32 no.4
• /
• pp.291-296
• /
• 1994

A bacterial strain NO. AM-28, showing proteolytic activity against defatted soybean was isolated from domestic soil. The isolated strain was identified as Aeromonas hydrophila by both the biochemical tests using API kit and the analysis of cellular fatty acid profile with MIDI system. The protease production from A. hydrophila AM-28 was highly enhanced when it was cultivated in the medium containing glycerol as a carbon source, tryptone or $(NH_4)_2HPO_4$ as a nitrogen source, and $CaCl_2$ as a mineral source. The optimal pH and temperature for the enzyme was 8.0 and $65^{\circ}C$, respectively. The enzyme was stable up to $55^{\circ}C$ and at pH values ranging from 7.0 to 13.0. The enzyme activity was inhibited by phenylmethylsulfonyl fluoride and EDTA, indicating that serine residue and metal ions be involved in enzyme activity.

• Korean Journal of Microbiology
• /
• v.24 no.2
• /
• pp.133-140
• /
• 1986

Extracts of CO-autotrophically grown cells of Acinetobacter sp. 1 were shown to use thionin, methylene blue, or 2,6-dichlorophenol-indophenol, but not NAD, NADP, FAD, or FMN, as electron acceptors for the oxidation of CO under strictly anaerobic conditions. The CO dehydrogenase (CO-DH) in the thes bacterium was found to be an inducible enzyme. The enzyme activity was determined by an assay based on the CO-dependent reduction of thionin. Maximal reaction rates were found at pH 7.5 and $60^{\circ}C$, and the Arrhenius plot revealed an activation energy of 6.1 kcal/mol(25.5kJ/mol). THe $K_m$ m/ for CO was $154{\mu}M$. Known metalchelating agents tested had no effects on the CO-DH activity. No divalent cations tested affect the enzyme activity significantly escept $Cu^{2+}$ which suppressed the activity completely. The enzyme was inhibited by glucose and succinate. The same extracts catalyzed oxidation of hydrogen gas and formate with thionin as electron acceptor. The CO-DH of Acinetobacter sp. 1 was to have no immunological relationship with that of Pseudomonas carboxydohydrogena.

• Microbiology and Biotechnology Letters
• /
• v.26 no.4
• /
• pp.345-351
• /
• 1998

In an effort to develop a potent system for the production of various dp (degree of polymerization) chitooligosaccharides, 32 enzymes or microbial systems were screened for chitosanolytic acitivity using chitosan as a substrate. The efficiency of each enzyme system was evaluated by the changes of turbidity and viscosity of chitosan solution, the amount of precipitate and the reducing sugar-producing activity in the enzymatic reaction mixture. Based on these assay methods for the chitosanase activity, Bacillus sp. P2l out of 32 screened systems showed highly potent endochitosanase, which was comparable with a commercially available enzyme (E7). Chitooligosaccharides of dp 3-7 were separated by TLC as major enzymatic reaction products, suggesting that the chitosanase from Bacillus sp. P2l be endo-splitting type.

• Microbiology and Biotechnology Letters
• /
• v.20 no.4
• /
• pp.401-408
• /
• 1992

The extracellular cholesterol oxidase produced from a soil microorganism HSL613 was purified and partially characterized. Through a series of purification procedures including concentration with CH2 concentrator, DEAE-cellulose column chromatography and gel filtration on Superose12, the purified enzyme was shown to have a specific activity of 108 units/mg protein giving 30.8-fold purification and final yield of 66%. The molecular weight of the enzyme was estimated to be 59,500 daltons by SDS-PAGE. The optimum temperature and pH for this enzyme were $50^{\circ}C$ and 6.0, respectively. The activity of the purified cholesterol oxidase was inhibited by $Ag^{2+}$, $Hg^{2+}$ and SDS.

• Korean Journal of Soil Science and Fertilizer
• /
• v.42 no.4
• /
• pp.307-316
• /
• 2009

Soil microbial activity and diversity are affected by organic sources applied to improve soil quality and fluctuate seasonally. We investigated the effects of municipal compost (MC), poultry litter (PL), and cover crops of spring oats and red clover (RC) on soil enzyme activities, and soil bacterial community-level physiological profiling (CLPP) in a Mexico silt loam in North Central Missouri, USA. Temporal patterns of these parameters were observed by periodic five soil sampling from spring to fall over a two year period. MC increased soil dehydrogenase (DH) activity consistently beginning about three months after MC application; fluorescein diacetate (FDA) hydrolytic activity significantly began to increase by the September of the first year but fluctuated during the following period. DH activity responded more directly to the amount or properties of organic residues in soils while FDA hydrolysis and CLPP were generally influenced by composition of organic sources, and enzyme activities and CLPP showed seasonal variation, which depended on organic sources and soil moisture. MC and cover crops may be useful organic sources for enhancing general soil microbial activity and altering soil microbial diversity, respectively. Because microbial activities and diversity are dynamic and subject to seasonal changes, the effects of organic amendments on these parameters should be investigated frequently during a growing season.