• Journal of Microbiology and Biotechnology
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• v.18 no.9
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• pp.1518-1521
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• 2008

Bacteria sense their population density and coordinate the expression of target genes, including virulence factors in Gram-negative bacteria, by the N-acylhomoserine lactones (AHLs)-dependent quorum sensing (QS) mechanism. In contrast, several soil bacteria are able to interfere with QS by enzymatic degradation of AHLs, referred to as quorum quenching. A potent AHL-degrading enzyme, AiiA, from Bacillus thuringiensis has been reported to effectively attenuate the virulence of bacteria by quorum quenching. However, little is known about the role of AiiA in B. thuringiensis itself. In the present study, an aiiA-defective mutant was generated to investigate the role of AHA in rhizosphere competence in the root system of pepper. The aiiA mutant showed no detectable AHL￢-egrading activity and was less effective for suppression of soft-rot symptom caused by Erwinia carotovora on the potato slice. On the pepper root, the survival rate of the aiiA mutant significantly decreased over time compared with that of wild type. Interestingly, viable cell count analysis revealed that the bacterial number and composition of E. carotovora were not different between treatments of wild type and the aiiA mutant. These results provide evidence that AHA can play an important role in rhizosphere competentce of B. thuringiensis and bacterial quorum quenching to Gram-negative bacteria without changing bacterial number or composition.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.550-555
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• 1995

A strain of Pseudomonas sp. isolated from soil was shown to produce a high level of extracellular endo-inulinase. In this work, the endo-inulinase gene (inu1) of the bacterial strain was cloned into the plasmid pBR322 by using EcoRI restriction endonuclease and E. coli HB101 as a host strain. One out of 7, 000 transformants obtained from the above cloning experiment formed a clear zone around its colony on the selective medium supplemented with 2.0% inulin after a prolonged incubation at 37$\circ$C and subsequent cold shock treatment. The functional clone was found to carry a recombinant plasmid (pKMG50) with a 3.7 kb genomic insert containing the genetic information for the inulinase activity. The inulinase from E. coli HB101/pKMG50 was proved to be an endo-acting enzyme and produced constitutively in the recombinant E. coli cells. Zymogram of the enzyme from the recombinant cells with inulin substrate indicated that the molecular mass of the active protein was 190 Kd, while that of the endo-inulinase from the Pseudomonas strain was 170 Kd. This size discrepancy suggested that the inulinase from the recombinant E. coli HB101 cells might be the initial product of translation, not the mature form produced in the strain of Pseudomonas sp..

• Microbiology and Biotechnology Letters
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• v.30 no.4
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• pp.332-338
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• 2002

A strain YB-4 producing the extracellular $\alpha$-galactosidase was isolated from soil, and has been identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. The partially purified $\alpha$-galactosidase was most active on paranitrophenyl-$\alpha$-D-galactopyranoside at pH 6.0 and 6$0^{\circ}C$. The enzyme retained 90% of its maximum activity between pH 4.0 and pH 10.0 after pre-incubation for 1 h. The enzyme was able to hydrolyze oligomeric substrates such as melibiose, raffinose and stachyose to liberate galactose residue, indicating that the $\alpha$-galactosidase of Steptomyces sp. YB-4 hydrolyzed $\alpha$-1,6 linkage.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.507-513
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• 1992

Serratia marecescens RH1 isolated from soil samples produced large amount of extracellular proteases. One of the genes encoding an extracellular protease form S. marcescens RH1 was cloned in Escherichia coli by shot gun cloning method. The cloned protease, SSP, was stably expressed by its own promoter and excreted into the extracellular medium from E. coli host (ORF) of 3.135 nucleotides corresponding to 1.045 amino acids (112 kDa). The nucleotide and deduced amino acid sequence of SSP showed high overall homology (88%) to one of the S. marcescens protease (27), but low homology to other serine protease families. The optimal pH and temperature of the enzyme were pH 9.0 and 45.deg.C respectively. The activity of protease was inhibited by phenylmethylsulfonyl fluoride (PMSF), which suggests that the enzyme is a serine protease.

• Journal of The Korean Society of Grassland and Forage Science
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• v.39 no.4
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• pp.298-302
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• 2019

Acidic soil significantly reduces crop productivity mainly due to aluminum (Al) toxicity. Alfalfa (Medicago sativa L.) roots were exposed to aluminum stress (Al₃⁺) in calcium chloride (CaCl₂) solution (pH4.5) and root growth, physiological and antioxidant enzyme responses were investigated. The root growth (length) was significantly inhibited after 48 h of aluminum stress imposition. Histochemical staining with hematoxylin indicated significant accumulation of aluminum in Al stress-treated root tissues. Histochemical assay were also performed to detect superoxide anion, hydrogen peroxide and lipid peroxidation, which were found to be more in root tissues treated with higher aluminum concentrations. The enzymatic activity of CAT, POD and GR in root tissues was slightly increased after Al stress treatment. The result suggests that Al stress alters root growth in alfalfa and induces reactive oxygen species (ROS) production, and demonstrates that antioxidant enzymes involved in detoxification of Al-mediated oxidative stress.

• Journal of Microbiology
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• v.37 no.4
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• pp.200-205
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• 1999

7-Aminocephalosporanic acid (7-ACA) is the initial compound in preparation of cephalosporin antibiotics widely used in clinical treatment. Bacteria producing glutaryl 7-ACA acylase, which convert cephalosporin C to 7-ACA, has been screened in soil samples. A bacterial strain exhibiting high glutaryl 7-ACA acylase activity, designated KAC-1, was isolated and identified as a strain of Pseudomonas diminuta by characterizing its morphological and physiological properties. The screening procedures include culturing on enrichment media containing glutaric acid, glutamate, and glutaryl 7-aminocephalosporanic acid as selective carbon sources. To enhance enzyme production, optimal cultivation conditions were investigated. This strain grew optimally at pH 7 to 9 and in temperatures of 20 to 40 C, but acylase production was higher when the strain was grown at 25 C. Glutaric acid, glutamate and glucos also acted as inducers for acylase production. In a jar fermenter culture, P. diminuta KAC-1 produce acylase in a growth-associated manner. The substrate specificity of KAC-1 acylase by cell extract showed that this enzyme had specificity toward glutaryl 7-ACA, glutaryl 7-ADCA, but not cephalosporin C.

• Mycobiology
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• v.51 no.5
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• pp.354-359
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• 2023

Tricholoma matsutake has been the most valuable ectomycorrhizal fungi in Asia because of its unique flavor and taste. However, due to the difficulty of artificial cultivation, the cultivation of T. matsutake has relied on natural growth in forests. To cultivate the T. matsutake artificially, microorganisms in fairy rings were introduced. In this study, we isolated 30 fungal species of microfungi from the soil of fairy rings. Among them, one single fungal strain showed a promoting effect on the growth of T. matsutake. The growth effect was confirmed by measuring the growth area of T. matsutake and enzyme activities including a-amylase, cellulase, and b-glucosidase. In comparison with control, microfungal metabolite increased the growth area of T. matsutake by 213% and the enzyme activity of T. matsutake by 110-200%. The isolated fungal strain was identified as Penicillium citreonigrum by BLAST on the NCBI database. The Discovery of this microfungal strain is expected to contribute to artificial cultivation of T. matsutake.

• Applied Biological Chemistry
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• v.38 no.6
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• pp.507-511
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• 1995

In an attempt to improve the productivity of ${\beta}-galactosidase$ from Bacillus sp. A1, which was isolated from soil and has remarkably higher transgalactosylation activity than lactose hydrolysis activity, a chemical mutation procedure using N-methyl-N'-nitro-N-nitrosoguanidine followed by selection was conducted. The final selection, designated as Bacillus sp. A4442, turned out to show a substantially increased enzyme productivity. Catabolite repression by glucose and lactose requirement as an inducer for the enzyme biosynthesis, which were shown in the parent strain, was markedly diminished; instead it was found out that galactose acts as another inducer. Because pH of medium, one of the most important factors for cell growth as well as enzyme production, is closely related with the sugar concentration during culture, it was kept in the optimum range of $6.5{\sim}7.5$; for this the initial glucose concentration was adjusted to be 0.5% which was thereafter maintained by the controlled pumping-in of lactose using the pH-stat technique. By doing so, we were able to increase the productivity of ${\beta}-galactosidase$ with high transgalactosylation activity up to $44\;unit/m{\ell}-broth$.

• Korean Journal of Soil Science and Fertilizer
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• v.37 no.4
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• pp.245-250
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• 2004

Farmers typically apply the dressed soil (coarse saprolite) for various reasons in the sloped upland with high altitude in Kangwon province. However, little researches on the impacts of application of dressed soil in uplands were conducted. Therefore, it is necessary to assess soil quality in this area and to study adverse effects on soil and water due to application of dressed soil. Coarse saprolite itself showed signiScantly poor chemical properties, Particularly P and organic matter contents were not enough for crops to grow. With respect to biological qualities such as enzyme activity and microbial population, coarse saprolite itself showed poor qualities. For example, bacterial population in coarse saprolite contains six times or ten times smaller populations. Based on survey at Jawoon-ri in Hongchon-gun, this region is susceptible for soil erosion due to massive amounts of coarse saprolite application, undesirably long slope length, etc. When weestimated soil loss, more than 40% of farming field in this region exceeded $11.2MT\;ha^{-1}\;yr^{-1}$. According to experiment by installing sediment basins. the sediment basin with up-down tillage and application with dressed soil had the highest soil loss and runofT, while the sediment basin with contour tillage and without soil dressing showed the lowest soil erosion and runoff.

• Journal of the Korean Society of Food Science and Nutrition
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• v.15 no.4
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• pp.32-39
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• 1986

A strain of Aspergillus niger CAD 1 which produces considerable amount of beta-galactosidase was selected from extracellular beta-galctaosidase producing fungi isolated from soil. Optimal conditions for the enzyme from Aspergillus niger CAD 1 were the growth in wheat bran supplemented with 0.5% skim milk powder at $30^{\circ}C$ for 72 hrs. The crude enzyme was purified 1,387 fold through DEAE-cellulosc and Sephadex G-100 chromatographr and its recovery was 6.2%, The optimal pH and temperature for the purified enzyme were pH 4.5 ana $45^{\circ}C$, respectively. The Km and Vmax on ONPG were $3.57{\times}10^3M$ and 33.0 unit/mg protein, whereas those on lacose were $83.3{\times}10^3M$and 15.33 unit/mg protein, respectively, The activation energy for the enzyme was 9,900 cal/mol and the enzyme had no metal ion requirement for its activity and stability. The hydrolysis of lactose in skim milk, 4.8% lactose solution and acidic whey were 65%, 70% and 78% after 10 hrs incubation at $45^{\circ}C$, when 182 units of the enzyme were used 50ml of the substrate solutions.