• Microbiology and Biotechnology Letters
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• v.23 no.3
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• pp.322-328
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• 1995

A thermophilic bacteria showing proteolytic activity against defatted soybean was isolated from soil. It was identified as Bacillus amyloliquefaciens based on its morphological and physiological characteristics. The Bacillus amyloliquefaciens NS 15-4 was cultivated at 50$\circ$C by rotary shaking in a medium containing defatted soybean. An extracellular protease from this strain was purified to homogeneity by ammonium sulfate precipitation, ion exchange, and hydrophobic interaction chromatographies. The molecular weight of the enzyme was estimated to be approximately 30,000 by SDS-PAGE and the N-terminal amino acid sequence of the enzyme was turned out to be AQSVPYGISQIKAPA. The optimum temperature and pH for the enzyme reaction were 60$\circ$C and 11, respectively, and its thermostability was increased by the addition of calcium ion. The enzyme was inactivated by phenylmethylsulfonylfluoride, suggesting it be a serine protease. Comparing with other commercial proteases, the enzyme showed relatively high proteolytic activity against defatted soybean, a water-insoluble protein substrate.

• Microbiology and Biotechnology Letters
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• v.24 no.6
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• pp.678-686
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• 1996

To improve the preservation of Kimchi, we isolated Lactobacillus plantarum lytic enzyme-producing strain from soil, and the enzyme was purified and characterized. From the observation of cultural and morpho- logical characteristics, the isolated strain was identified as Metarrisium anisopliae (Metschn.) Sorok. The enzyme was purified to 75-folds with 40% yields through affinity adsorption and CM-Sephadex C-50 column chromatog- raphy. The optimum pH and temperature for lytic activity are 4.0 and 40$\circ$C, respectively, and the enzyme acitvity is stable between pH 3.0 and 9.0, and up to 50$\circ$C. The enzyme is a monomeric protein with molecular weight of 40,000 daltons by SDS-PAGE and gel filtration. The enzyme is endopeptidase which breaks the peptide linkage of Lactobacillus plantarum peptidoglycan. The lytic action spectra confirmed that Leuconostoc mesenteroides, a useful strain for the fermentation of Kimchi, is not lysed by the enzyme. The enzyme activity is inhibited by N-bromosuccinimide (NBS), which probably indicates the involvement of tryptophan residue in active site of the enzyme, and also inhibited by Ag$^{+}$. The amino acid composition analysis showed that the enzyme contains more acidic amino acids than basic ones, and composition of alanine, glycine, proline and tyrosines was very high.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.104-114
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• 2021

Petroleum-contaminated soil is considered among the most important potential anthropogenic atmospheric methane sources. Additionally, various rhizoremediation factors can affect methane emissions by altering soil ecosystem carbon cycles. Nonetheless, greenhouse gas emissions from soil have not been given due importance as a potentially relevant parameter in rhizoremediation techniques. Therefore, in this study we sought to investigate the effects of different plant and soil amendments on both remediation efficiencies and methane emission characteristics in diesel-contaminated soil. An indoor pot experiment consisting of three plant treatments (control, maize, tall fescue) and two soil amendments (chemical nutrient, compost) was performed for 95 days. Total petroleum hydrocarbon (TPH) removal efficiency, dehydrogenase activity, and alkB (i.e., an alkane compound-degrading enzyme) gene abundance were the highest in the tall fescue and maize soil system amended with compost. Compost addition enhanced both the overall remediation efficiencies, as well as pmoA (i.e., a methane-oxidizing enzyme) gene abundance in soils. Moreover, the potential methane emission of diesel-contaminated soil was relatively low when maize was introduced to the soil system. After microbial community analysis, various TPH-degrading microorganisms (Nocardioides, Marinobacter, Immitisolibacter, Acinetobacter, Kocuria, Mycobacterium, Pseudomonas, Alcanivorax) and methane-oxidizing microorganisms (Methylocapsa, Methylosarcina) were observed in the rhizosphere soil. The effects of major rhizoremediation factors on soil remediation efficiency and greenhouse gas emissions discussed herein are expected to contribute to the development of sustainable biological remediation technologies in response to global climate change.

• Applied Biological Chemistry
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• v.22 no.4
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• pp.217-220
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• 1979

Effects of nitrogen containing herbicides, Asulam(methyl-(4-aminobenzenesulphonyl)-carbamate), dimetametryne (2-methyl-4-ethylamino-6-(1,2-dimethyl propylamino)-S-triazine) and linuron (3-(3,4-dichlorophenyl)-1-metoxy-1-methyl urea) at rates of 0.5,2,4 mg/100g soil on urease activity were studied in urea added and unadded soil by incubating at $25{\pm}1^{\circ}C$ for 80 days. The enzyme activity was somewhat suppressed by asulam and dimetametryne in soils treated with urea. Unlike the above results, the enzyme activity in soil treated with linuron was kept higher as compared with that in soil treated with urea only.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2005.04a
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• pp.171-174
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• 2005

Suspected carcinogen, TCE and PCE, are the most common groundwater pollutants extensively used as a solvent and degreaser. In this study, oxygenases were immobilized in Ca-alginate and chitosan bead. TCE degradation by the immoblized enzyme beads were measured for various size, enzyme addition volume and TCE contact time. The degradation was decreased as increasing the bead size. For overnight , more than 20% of TCE was degraded. The variation of enzyme activity was tested for the repeated use of enzyme beads.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.102-110
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• 1991

Alkalophilic Bacillus sp. YJ-451, which was isolated from soil at several area in Korea, produced a novel type of bacteriolytic enzyme (cell wall peptidoglycan hydrolase) extracellulary. The cell wall hydrolytic activity was identified as a clear zone on sodium dodecyl sulfate polyacrylamide gel electrophoresis containing 0.2% (w/v) cell wall of Bacillus sp. as substrate. This enzyme was successively purified 66 fold with 3.2% yield in culture broth by ammonium sulfate precipitation, CM-cellulose column chromatography, and gel filtration, followed by hydroxylapatite column chromatography. The molecular weight of the purified enzyme was estimated to be 27,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum pH and temperature for the activity of the enzyme were pH 10.0 and $50^{\circ}C$, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to $40^{\circ}C$. Among the microorganisms used in this experiment the enzyme was active against most of gram negative strains and the genus Bacillus such as B. megaterium, B. licheniformis, B. circulans, B. pumilus, B. macerans, B. polymyxa. The release of dinitrophenylglutamic acid but not reducing group from cell wall peptidoglycan digested by the enzyme suggested that the enzyme is a kind of peptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan.

• Journal of Soil and Groundwater Environment
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• v.29 no.1
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• pp.51-62
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• 2024

A study was performed to investigate the positive impacts of humic substances (HS) on the growth of green barley, a type of green manure plant. The study was conducted in a pot culture using two different types of reclaimed soils that had been treated by land farming (DDC) and thermal desorption (YJ) methods, respectively. The experimental conditions consisted of three treatments: plant only (P), plant plus 2% HS, and no plant (control). After 89 days of culture in a controlled growth chamber, the growth of spring barley and activity of seven soil enzymes were measured. The results indicated that the addition of HS had a substantial (p<0.10) positive effect on shoot biomass in both types of soil. Furthermore, the addition of HS notably (p<0.05) enhanced all seven soil enzyme activities in both soils. Both the aboveground and belowground parts of barley plants were returned to soil and aged for 10 weeks in the same growth chamber, which resulted in notable enhancement in soil health indicators. These improvements included an increase in organic matter, a drop in bulk density, and an increase in the activity of seven different soil enzymes. When lentil seeds were planted in the aged soils, the development of the seedlings was more vigorous than that in the control in both soils, although allelopathy of barley suppressed lentil germination in soil with pH 7.0 but not in soil with pH 8.5.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.360-364
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• 2006

Fibrin clots of blood vessels are one of the serious factor caused cardiovascular disease. The development of a antithrombotic and thrombolysis solvent is necessary to prevent and treat these diseases. It has been reported that a strong fibrin-specific fibrinolytic enzyme was produced from a Korean fermented soybean paste similar to Japanese miso. We have been screened the known or novel fibrinolytic enzymes by activity-based and sequence-based screening from soil DNA metagenome library containing all kinds of environmental genomic DNA. The activity-based screening was determined the protease activity on 0.5% skim milk. For sequence-based screening, we designed a set of primer expanding gene sequence of fibrinolytic enzyme, performed PCR and selected clones showing the expected size of amplicons from metagenome library. Transformation of the gene encoding fibrinolytic enzyme was carried out with commercial vectors and their transformants were selected. Finally, we found 15 positive clones from metagenome library. Then each of sequences were analyzed and identified as similar or known the clones of nattokinase. We are going to perform full sequence of each clones, ligate with expression vector, transform into competent cells and then determine activity of expressed enzymes.

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

• Journal of Microbiology and Biotechnology
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• v.26 no.10
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• pp.1717-1722
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• 2016

A novel phytase of Acidobacteria was identified from a soil metagenome, cloned, overexpressed, and purified. It has low sequence similarity (<44%) to all the known phytases. At the optimum pH (2.5), the phytase shows an activity level of 1,792 μmol/min/mg at physiological temperature (37℃) and could retain 92% residual activity after 30 min, indicating the phytase is acidophilic and acidostable. However the phytase shows poor stability at high temperatures. To improve its thermal resistance, the enzyme was redesigned using Disulfide by Design 2.0, introducing four additional disulfide bridges. The half-life time of the engineered phytase at 60℃ and 80℃, respectively, is 3.0× and 2.8× longer than the wild-type, and its activity and acidostability are not significantly affected.