• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.588-596
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• 2018

An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

• Microbiology and Biotechnology Letters
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• v.17 no.2
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• pp.142-147
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• 1989

Six plasmids of B. thuringiensis serovar. kurstaki HD73 were detected, with approximate sizes of 7.4, 7.8, 8.1, 11.3, and 75 Kb, as well as a low copied plasmid of similar length to 75 Kb. Partially cured mutants from B. thuringiensis HD73 were obtained either by the treatment of the curing agent, ethidium bromide(0.02 $\mu\textrm{g}$/$m\ell$) or by spontaneous curing, Acrystalliferous mutants(Cry$^-$) were identified by microscopic observation and immunoblotting with polyclonal antibody against 133 KD deltaendotoxin of HD73. Ten Cry$^-$ mutants were found to be lack of 75 Kb plasmid. These results implicated that this plasmid was associated with delta-endotoxin production, After isolating the mutants, we streaked them on potato dextrose agar, spizizen casamino acid glucose, starch agar, and nutrient agar. Only on starch agar medium did morphologies of Cry$^-$ appear translucent and light greyish. On the other hand, the mutants of B. thuringiensis isolated from Korean soil, designated KBS722, were obtained by the treatment of novobiocin (3 $\mu\textrm{g}$/$m\ell$). Acrystalliferous mutants of KBS722 were less translucent than HD73 mutants' only on nutrient agar medium. Compared the plasmid profile of the mutants with delta-endotoxin production, the results seemed to indicate that the insecticidal protein gene of B. thuringiensis isolates KBS722 located on about 225 Kb plasmid DNA.

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.580-586
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• 1989

Penicillium sp. CB-20 was selected for strong polygalacturonase activity among various strains of molds found in soil. The optimum pH for the enzyme activity was 5.0 and optimum temperature was 4$0^{\circ}C$. The enzyme was relatively stable in acidic condition and unstable by heat treatment. The activation energy, Km and V$_{max}$ for the polygalacturonase were 2.499 Kcal/mol, 2.13$\times$10$^{-2}$mol/l, and 104.17 $\mu$mol/min. The activity of polygalacturonase was inhibited by Ag$^{+}$, Cu$^{++}$, Pb$^{++}$, Fe$^{+++}$, $Ca^{++}$, Na$^+$, Mn$^{++}$. The enzyme can be inactivated by the treatment ethylenediamintetra acetic acid, 2,4-dinitrophenol and $H_2O$$_2$. The results indicate the possible involvement of histidine, chelate and terminal amino group as active site. The enzyme was endo-type polygalacturonase.

• KSBB Journal
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• v.14 no.6
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• pp.718-723
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• 1999

In order to treat of alkaline dye-processing wastewater, alkalophilic strains biodegrading azo dye, Acid red 1, is isolated from natural system, and optimal culture conditions are examined using response surface analysis, statistical analysis system program. 15 different species which grow in alkaline culture media are isolated from the effluent and river soil discharged from wastewater treatment plant in dye industrial complex. One strain which has the best decolorization efficiency is chosen, and named as AR-1. The result of the examination of carbon, nitrogen and phosphorus sources which have influence on growth and decolorization reveals that optimum carbon, nitrogen and phosphorus sources are 1.0% fructose, 1.0% polypeptone, 1.0% yeast extract and 0.5% $K_2HPO_4$, respectively. In order to optimize of biodegradation conditions of dye by response surface analysis, the characteristics of decolorization and cell growth according to culture temperature and time are monitered. The result shows that the one is optimum 34.77$^{\circ}C$ for 12.97 hours; the other at 34.73$^{\circ}C$ for 12.96 hours. While, optimal conditions of culture that satisfy both cell growth and decolorization are the temperatures from 32.86$^{\circ}C$ to 36.36$^{\circ}C$ and the period of 10.96 to 15.75 hours, respectively.

• Journal of Korean Society for Atmospheric Environment
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• v.13 no.5
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• pp.371-381
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• 1997

Precipitation sampls were collected in Chonju-city during October 1994 to September 1995 and were analysed for major ions (N $a^{+}$, $K^{+}$, $Ca^{2+}$, $Mg^{2+}$, C $l^{[-10]}$ , NO/$_3$, S $O_4$$^{2-}$) and trace metals (Al, Cd, Ni, Pb, Sr, Zn) in addition to pH, in order to understand the chemical characteristics of acid rain and to estimate the origin of the determined ions. Most rain showed a neutral or alkaline character, and only 35% had a pH lower than 5.6. S $O_4$$^{2-}$ and N $O_3$$^{[-10]}$ are identified as the primary contributors to precipitation acidity in this region. Neutralization of precipitation acidity occurs as a result of the dissolution of alkaline compounds containing $Ca^{2+}$, $Mg^{2+}$ and $K^{+}$. S $O_4$$^{2-}$ and N $O_3$$^{[-10]}$ precipitation concentrations exhibit a seasonal pattern in which higher concentrations are observed during spring months and lower concentrations during summer months. However, the seasonal behavior of $H^{+}$ concentrations differs from this pattern, in that the highest concentrations occur during autumn months, owing to the different influence of neutralization processes. In all rain, S $O_4$$^{2-}$ concentration exceeded NO/$_3$$^{[-10]}$ concentration. The contribution of maritime sources to the total S $O_4$$^{2-}$ concentration was very low or negligible. For rain strongly affacted by yellow sand, $Ca^{2+}$, $Mg^{2+}$ and $K^{+}$ ions show a sharp increase in concentration, reflecting the increased amount of dust and soil suspended in atmosphere. At the same time, S $O_4$$^{2-}$ and N $O_3$$^{[-10]}$ concentrations are at their highest levels while $H^{+}$ values are not comparably elevated, presumably beacause much of the acidity has been neutralized by alkaline substances. The seasonal variance of trace metal concentrations in rainwater is similar to that of major cations. The annual wet flux of acidic pollutants and trace metals wat calculated to be as follows: N $O_3$$^{[-10]}$ ; 2.32 g/$m^2$, S $O_4$$^{2-}$, 5.34 g/$m^2$, Al; 6.30 mg/$m^2$, Cd; 0.62 mg/$m^2$, Ni; 4.08 mg/$m^2$, Pb: 9.76 mg/$m^2$, Sr; 5.94 mg/$m^2$, Zn; 111 mg/$m^2$./$m^2$.

• Journal of Microbiology
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• v.43 no.3
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• pp.285-294
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• 2005

A bacterial strain M4-7 capable of degrading various polyesters, such as poly$(\varepsilon-caprolactone)$, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3-hydroxyoctanoate), and poly(3-hydroxy-5-phenylvalerate), was isolated from a marine environment and identified as Pseudomonas alcaligenes. The relative molecular mass of a purified extracellular medium-chain-length poly(3-hydroxyalkanoate) (MCL-PHA) depolymerase $(PhaZ_{palM4-7})$ from P. alcaligenes M4-7 was 28.0 kDa, as determined by SDS-PAGE. The $PhaZ_{palM4-7}$ was most active in 50 mM glycine-NaOH buffer (pH 9.0) at $35^{\circ}C$. It was insensitive to dithiothreitol, sodium azide, and iodoacetamide, but susceptible to p-hydroxymercuribenzoic acid, N-bromosuccinimide, acetic anhydride, EDTA, diisopropyl fluorophosphate, phenylmethylsulfonyl fluoride, Tween 80, and Triton X-100. In this study, the genes encoding MCL-PHA depolymerase were cloned, sequenced, and characterized from a soil bacterium, P. alcaligenes LB19 (Kim et al., 2002, Biomacro-molecules 3, 291-296) as well as P. alcaligenes M4-7. The structural gene $(phaZ_{palLB19})$ of MCL-PHA depolymerase of P. alcaligenes LB19 consisted of an 837 bp open reading frame (ORF) encoding a protein of 278 amino acids with a deduced $M_r$ of 30,188 Da. However, the MCL-PHA depolymerase gene $(phaZ_{palM4-7})$ of P. alcaligenes M4-7 was composed of an 834 bp ORF encoding a protein of 277 amino acids with a deduced Mr of 30,323 Da. Amino acid sequence analyses showed that, in the two different polypeptides, a substrate-binding domain and a catalytic domain are located in the N-terminus and in the C-terminus, respectively. The $PhaZ_{palLB19}$ and the $PhaZ_{palM4-7}$ commonly share the lipase box, GISSG, in their catalytic domains, and utilize $^{111}Asn$ and $^{110}Ser$ residues, respectively, as oxyanions that play an important role in transition-state stabilization of hydrolytic reactions.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.204-210
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• 2006

Ethyl (S)-4-chloro-3-hydroxybutyrate is a useful intermediate for the synthesis of Atorvastatin, a chiral drug to hypercholesterolemia. In this research, two 4-chloro-3-hydroxybutyro-nitrile-degrading strains were isolated from soil sample. They were identified as Rhodococcus erythropolis strains by 16S rRNA analysis. The nitrile-degrading enzyme(s) were suggested to be nitrile hydratase and amidase rather than nitrilase from the result of thin layer chromatography analysis. The corresponding genes were obtained by PCR cloning method. The predicted protein sequences had identities more than 96% with nitrile hydratase ${\alpha}-subunit$, nitrile hydratase ${\beta}-subunit$, and amidase of R. erythropolis. The 4-chloro-3-hydroxybutyronitrile-hydrolyzing activities in both strains were increased dramatically by ${\varepsilon}-caprolactam$ which was known as good inducer for nitrile hydratase. Both intact cells and cell-free extract could hydrolyze the nitrile compound. So, the intact cell and the enzymes could be used as potential biocatalyst for the production of 4-chloro-3-hydroxybutyric acid.

• Korean Journal of Weed Science
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• v.31 no.3
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• pp.240-249
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• 2011

The potential of juglone a plant naphthoquinone as a natural herbicide on new target, 7-keto-8-amino pelargonic acid synthetase (KAPAS) in the early step of biotin biosynthesis pathway, was performed in vitro and in vivo. Juglone effectively inhibited KAPAS activities in vitro and the $IC_{50}$ was $9.5{\mu}M$. Foliar application of juglone showed very good herbicidal activity to the eight-tested weed species. Among them, Solanum nigrum was completely controlled at a concentration of $250{\mu}g\;mL^{-1}$ with main symptoms of desiccation or burndown. Digitaria sanguinalis and Aeschynomene indica were also sensitive to juglone treatment. All eight weed species were controlled by 90~100% at a concentration of $500{\mu}g\;mL^{-1}$. However, soil application of juglone to Digitaria sanguinalis did not show any herbicidal symptoms. Cellular leakage from cucumber leaf squares treated with juglone increased depending on the concentrations increased from 6.25 to $100{\mu}M$ after 24 hours incubation with or without light. However, chlorophyll loss in cucumber leaf squares was negligible. Biotin supplements significantly rescued the inhibition of germination rate of Arabidopsis thaliana seeds previously inhibited by the juglone. Our results suggest that the juglone is a possible environmental friendly herbicide candidate with a new target KAPAS inhibiting activity.

• Journal of Life Science
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• v.23 no.10
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• pp.1295-1303
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• 2013

According to facts revealed up until the present, there are a total of 68 known phyla on earth, including 55 phyla of bacteria and 13 phyla of archaea. The human large intestine has 9 phyla of microorganisms, which is a relatively lower diversity compared to the general environments of soil or sea. The diversity of intestinal microorganisms is affected by the characteristics of the host (genetic background, sex, age, immune system, and gut motility), the diet (non-digestible carbohydrates, fat, prebiotics, probiotics), and the intake of antibiotics, which in turn have an effect on energy storage processes, gene expressions, and even metabolic diseases like obesity. Probiotics are referred to as living microorganisms that improve the intestinal microbiota and contribute to the health of the host; in addition, probiotics usually comprise lactic acid bacteria. Recently, bacteriotherapy using probiotics has been utilized to treat sicknesses like diarrhea and irritable bowel syndrome. Prebiotics are a food ingredient which can selectively adjust intestinal microorganisms and which comprise inulin, fructooligosaccharides, galactooligosaccharides, and lactulose. In recent days, attention has been paid to the use of dietary cellulose in the large intestine and the production of short chain fatty acids (short-chain fatty acids) in relation to obesity and anticancer. More research into microorganisms in the large intestine is necessary to identify specific microorganism species, which are adjusted by diverse non-digestible carbohydrates, prebiotics, and probiotics in the large intestine and to understand the connection between sicknesses and metabolites like short chain fatty acids produced by these microorganism species.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.195-199
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• 1998

AH alkalophilic Bacillus SP. AIk-7, isolated from soil, produced ethylene. The characteristics of this microorganism is the ability to grow well under the alkaline condition, at pH 10.3. This strain is similar to Bacillus alkalophilus in terms of morphological, physiological and biological characteristics. In observation of relationship of cell growth and ethylene production according to incubation times, the ethylene synthesis mostly occur from the late exponential phase to the death phase of growth. The purpose of this paper is to study the effects of various substrates on the biosynthesis of ethylene in the intact cell and the cell-free system by the Bacillus sp. AIk-7. In both intact cell and cell-free extract, optimum conditions for ethylene production was achieved at pH 10.3 and 3$0^{\circ}C$. Ethylene was effectively produced from L-Met and 1-aminocyclopropane-1-carboxylic acid (ACC). In this case, ACC as the substrate on ethylene production were two fold higher than L-met at each concentration of substrates. On the other hand, the cell-free ethylene-forming system was used as a tool for the elucidation of the biochemical reaction involved in the formation of ethylene by Bacillus sp. AIk-7. Ethylene production in the cell-free system required the presence of manganese and cobalt ion to be stimulated a little. The result obtained in this work suggests that L-met and ACC may be a precursor more directly related to bacterial ethylene production than any other substrates tested.