• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.574-582
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• 1992

Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.

• KSBB Journal
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• v.14 no.4
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• pp.422-428
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• 1999

A transformat Alcaligence eutrophus GA5 harboring phbC gene from A. latus was cultivated for production of Poly(3-hydroxybutyrate-4-hydroxybutyrate)[P(3HB-4HB)] containing high molar fraction of 4-hydroxybutyrate(4HB)] containing high molar fraction of 4-hydroxybutyrate(4HB). Transformation did not influenced significantly on total cell growth, on total cell growth, concentration, and content of P(3HB-4HB), however, significantly influenced on 4HB molar fraction in P(3HB-4HB) increasing from 12.3 to 23.5 mol% after 48 h cultivation in two-stage using 1.0%(W/V) of ${\gamma}$-butyrolactone as a precursor compare to parent strain. Above increment may be due to the accelerated polymerization between 3HB and 4HB converted from precusor compound by amplified phbC gene. Citrate increased remarkbly total cell mass and P(3HB-4HB) concentration, but did not influenced on the molar fraction of 4HB, meanwhile, magnesium ion influenced on P(3HB-4HB) concentration and 4HB molar fraction significantly. The two-stage cultivation method was modified, in such a way minimizing P(3HB) accumulated inside of cell grown at first-stage, consquently, 26.3% of P(3HB-4HB) containing 61.0 mol% of 4HB fraction was obtained after 72hr. Furthermore, semi-homopolymeric P(4HB) containing 92.0 mol% of 4Hb was obtained, and its structure was confirmed by $^1$H-NMR.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.154-160
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• 1986

In order to develope interspecific fusion of the genus Cellulomonas capable of assimilation cellulose, the optimun conditions for the protoplast formation was investigated to examine the susceptibility of cell wall, between different species of the same genus using scanning electron microscope. The variation in the susceptibilities of Cellulomonas sp. CS 1-1 and C. flavigena to lysozyme treatment were considerably remarkable, although they belong to the same genus. The rate of protoplast formation of CS1-1 was 99.9% being treated with lysozyme $(100{\mu}g/ml)$ for 30 minute and that of C. flavigena was about 80% being treated at the concentration of $600{\mu}g/ml$ of lysozyme for 6 hours. The susceptibility of cell wall to the lysozyme treatment on protoplast formation of the strain, CS1-1 seems not to be depend on the cultural periods of cells. On the contrary, that of C. flavigena was considerably depend on the periods. Cells of C. flavigena at mid exponential phase could be more efficiently converted to protoplast cells than those at late exponential phase be done. The rate of the protoplast formation was 95%, when cells of C. flavigena at mid exponential phase were treated with lysozyme $600{\mu}g/ml$ for 6 hours and observed by SEM. In the evalution of protoplast formation of the CS1-1 results of counting method in plate after osmotic shock treatment were similar to the results of the direct observation method by means of SEM. But in the case of C. flavigena the latter method was much more reliable than the former, because the differences between the number of spheroplasts and protoplasts were not able to figure out on conuting the number of protoplast after osmotic shock tretment.

• Korean Journal of Soil Science and Fertilizer
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• v.42 no.1
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• pp.29-36
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• 2009

Fourteen bacterial strains were isolated from crop rhizosphere and identified as phosphate solubilizing bacteria (PSB) by 16S rRNA analysis. Only 3 strains exhibited a strong ability to solubilize insoluble phosphate in agar medium containing a hydroxyapatite. The rates of P solubilization by isolates were ranged from 200 and $2300\;mg\;L^{-1}$, which are inversely correlated with pH in culture medium. Furthermore, HPLC analyses reveal the production of organic acid from the culture filtrates of PSB. Among these, strain Acinetobacter sp. released only gluconic acid, Pseudomonas orientalis produced gluconic acid which was subsequently converted into 2-ketogluconic acid, and Enterobacter asburiae released acetic acid and succinic acid. On the other hand, P. orientalis and E. asburiae released $372\;mg\;L^{-1}$ and $191\;mg\;L^{-1}$ of IAA into broth culture, respectively, while Acinetobacter sp. did not produce IAA. Furthermore, in vivo study showed that plant growth promoting effect by bacteria generally seemed to be increased IAA production and phosphate solubilization.

• Journal of Microbiology and Biotechnology
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• v.31 no.2
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• pp.259-271
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• 2021

Many bacteria metabolize aromatic compounds via catechol as a catabolic intermediate, and possess multiple genes or clusters encoding catechol-cleavage enzymes. The presence of multiple isozyme-encoding genes is a widespread phenomenon that seems to give the carrying strains a selective advantage in the natural environment over those with only a single copy. In the naphthalene-degrading strain Pseudomonas putida ND6, catechol can be converted into intermediates of the tricarboxylic acid cycle via either the ortho- or meta-cleavage pathways. In this study, we demonstrated that the catechol ortho-cleavage pathway genes (catBICIAI and catBIICIIAII) on the chromosome play an important role. The catI and catII operons are co-transcribed, whereas catAI and catAII are under independent transcriptional regulation. We examined the binding of regulatory proteins to promoters. In the presence of cis-cis-muconate, a well-studied inducer of the cat gene cluster, CatRI and CatRII occupy an additional downstream site, designated as the activation binding site. Notably, CatRI binds to both the catI and catII promoters with high affinity, while CatRII binds weakly. This is likely caused by a T to G mutation in the G/T-N11-A motif. Specifically, we found that CatRI and CatRII regulate catBICIAI and catBIICIIAII in a cooperative manner, which provides new insights into naphthalene degradation.