• Journal of Microbiology and Biotechnology
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• v.28 no.9
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• pp.1502-1510
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• 2018

Organic solvent-tolerant (OST) enzymes are widely applied in various industries for their activity and stability in organic solvents, for their higher substrate solubility, and for their greater stero-selectivity. However, the criteria for identifying OST enzymes largely remain undefined. In this study, we compared the amino acid composition of 19 OST esterases with that of 19 non OST esterases. OST esterases have increased the ratio of Ala and Arg residues and decreased the ratio of Asn, Ile, Tyr, Lys, and Phe residues. Based on our amino acid composition analysis, we cloned a carboxylesterase (EstSP2) from a psychrophilic bacterium, Sphingomonas glacialis PAMC 26605, and characterized its recombinant protein. EstSP2 is a substrate specific to p-nitrophenyl acetate and hydrolyzed aspirin, with optimal activity at $40^{\circ}C$; at $4^{\circ}C$, the activity is approximately 50% of its maximum. As expected, EstSP2 showed tolerance in up to 40% concentration of polar organic solvents, including dimethyl sulfoxide, methanol, and ethanol. The results of this study suggest that selecting OST esterases based on their amino acid composition could be a novel approach to identifying OST esterases produced from bacterial genomes.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.756-762
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• 2001

As a component for a recirculating aquaculture system, a new strain of denitrifying bacterium was isolated from municipal sewage. The isolate was motile by means of one polar flagellum, catalase-positive, and a Gram-negative rod-shaped cell measuring $0.5-0.6{\mu}m$ in width and $1.3-1.9{\mu}m$ in length. The isolate was identified as Pseudomonas fluorescens and produced dinitrogen gas via the reduction of nitrate. The optimal growth conditions (pH, temperature, carbon source, and C/N ratio) of the isolate were found to be 6.8, $30^{\circ}C$, malate, and 3, respectively. Under optimal growth conditions of P. fluorescens, dinitrogen gas was first detected in the exponential growth phase, then a small amount of nitrite was developed and converted to dinitrogen gas in the stationary phase. Pseudomonas fluorescens cells were immobilized in modified polyvinyl alcohol (PVA) gel beads, and the maximum denitrification rate was measured as $36.6 {\mu}lN_2h^-1$ per bead with an optimum cell loading of $20mg {\mu}l^-1$ and $2\%$ sodium alginate added to the PVA gel. The operating stability of the modified PVA gel beads remained unchanged for up to 43 repeated batches.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1300-1305
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• 2017

A rod shaped, aerobic, Gram-stain-negative, and motile bacterium, strain $AR2^T$, was isolated from a water sample of Yeongsan river, Republic of Korea. Strain $AR2^T$ clustered closely with the members of the genus Chitinimonas and showed the highest 16S rRNA gene sequence similarity with Chitinimonas prasina $LY03^T$ (96.4%), Chitinimonas viridis $HMD2169^T$ (96.4%), Chitinimonas taiwanensis $cf^T$ (96.2%), and Chitinimonas koreensis $R2A43-10^T$ (94.2%). The predominant fatty acids of strain $AR2^T$ were identified to be summed feature 3 (comprising $C_{16:1}{\omega}7c$ and/or $C_{16:1}{\omega}6c$), $C_{16:0}$, and $C_{10:0}3-OH$. Diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine were found to be the major polar lipids. The genomic DNA G+C content was 60.4 mol%. Based on the polyphasic characterization, the isolated strain $AR2^T$ is described as a representative of a novel species in the genus Chitinimonas, for which the name Chitinimonas naiadis sp. nov. (type strain =$AR2^T$ =KCTC $42755^T$ =JCM $31504^T$) is proposed.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.384-390
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• 2021

A Gram-negative, aerobic, motile by gliding, and rod-shaped marine bacterium, designated CE67^T was isolated from the marine sponge Callyspongia elegans on Biyang-do in Jeju Island. The CE67^T strain grew optimally at 25℃, pH 7.5, and in the presence of 2-3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain CE67^T was related to the genus Aureivirga and had the highest 16S rRNA gene sequence similarity to the Aureivirga marina VIII.04^T type strain (96.3%). The primary fatty acids (>10%) of strain CE67^T were iso-C_15:0 (35.3%) and iso-C_17:0 3OH (21.8%). The polar lipid profile of strain CE67^T contained phosphatidylethanolamine, unidentified aminolipids, and unidentified lipids. The predominant menaquinone was MK-6. The DNA G+C content was 29.1 mol%. Based on the polyphasic taxonomic analysis, strain CE67^T was determined to be a representative novel species of the genus Aureivirga for which we propose the name Aureivirga callyspongiae sp. nov., whose strain type is CE67^T (=KCTC 42847^T=JCM 34566^T).

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.324-330
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• 2014

A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.

• Microbiology and Biotechnology Letters
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• v.43 no.4
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• pp.314-321
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• 2015

Agar-hydrolyzing bacteria were isolated from the coastal sea water of Jeju Island. One isolate, designated as S4, was selected for further study. The S4 cells were Gram-negative and rod-shaped with smooth beige surfaces and single polar flagellum. Cells were grown at $15-42^{\circ}C$, 0.5-5% (w/v) NaCl, between pH 6.0 and 9.0, and in media containing 0.5-5% (w/v) NaCl. The G+C content was 49.93 mol%. The major fatty acids (>15%) were $C_{18:1}{\omega}7c$, $C_{16:0}$ and Summed feature 3 (comprising $C_{16:1}{\omega}7c/iso-C_{15:0}$ 2-OH). Based on 16S rRNA sequencing and biochemical and chemotaxonomic characteristics, the strain was designated as Vibrio sp. S4. In liquid culture supplemented with 0.1% agar the cell density and agarase activity reached a maximum level in 72 h, while agarase activity in the culture without agar was negligible, implying agarose expression is induced by agar. The optimum pH and temperature for the extracellular crude agarase of S4 were 7.0 and $45^{\circ}C$, respectively. However, it also exhibited 98.6% and 87.6% at $40^{\circ}C$ and $50^{\circ}C$, respectively, of the maximum activity seen at $45^{\circ}C$. The crude agarase hydrolyzed agarose into (neo)agarotetraose and (neo)agarohexaose.

• Journal of Microbiology and Biotechnology
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• v.33 no.2
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• pp.188-194
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• 2023

Microbacterium elymi KUDC0405^T was isolated from the rhizosphere of Elymus tsukushiensis from the Dokdo Islands. The KUDC0405^T strain was Gram-stain-positive, non-spore forming, non-motile, and facultatively anaerobic bacteria. Strain KUDC0405^T was a rod-shaped bacterium with size dimensions of 0.3-0.4 × 0.7-0.8 ㎛. Based on 16S rRNA gene sequences, KUDC0405^T was most closely related to Microbacterium bovistercoris NEAU-LLE^T (97.8%) and Microbacterium pseudoresistens CC-5209^T (97.6%). The dDDH (digital DNA-DNA hybridization) values between KUDC0405^T and M. bovistercoris NEAU-LLE^T and M. pseudoresistens CC-5209^T were below 17.3% and 17.5%, respectively. The ANI (average nucleotide identity) values among strains KUDC0405^T, M. bovistercoris NEAU-LLE^T, and M. pseudoresistens CC-5209^T were 86.6% and 80.7%, respectively. The AAI (average amino acid identity) values were 64.66% and 64.97%, respectively, between KUDC0405^T and its closest related type strains. The genome contained 3,596 CDCs, three rRNAs, 46 tRNAs, and three non-coding RNAs (ncRNAs). The genomic DNA GC content was 70.4%. The polar lipids included diphosphatydilglycerol, glycolipid, phosphatydilglycerol, and unknown phospholipid, and the major fatty acids were anteiso-C_17:0 and iso-C_16:0. Strain KUDC0405^T contained MK-12 as the major menaquinone. Based on genotypic, phylogenetic, and phenotypic properties, strain KUDC0405^T should be considered a novel species within the genus Microbacterium, for which we propose the name M. elymi sp. nov., and the type strain as KUDC0405^T (=KCTC 49411^T, =CGMCC1.18472^T).

• Journal of Microbiology and Biotechnology
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• v.33 no.10
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• pp.1292-1298
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• 2023

PAMB 00755^T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20℃, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755^T was most closely related to Sphingomonas chungangi MAH-6^T (97.7%) and Sphingomonas polyaromaticivorans B2-7^T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755^T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C_18:1ω7c and/or C_18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755^T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755^T as the type strain (= KCTC 92781^T = GDMCC 1.3779^T).

• Journal of Microbiology and Biotechnology
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• v.29 no.12
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• pp.1969-1974
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• 2019

A Gram-staining-negative, aerobic, light brown pigment bacterium, designated strain CE80^T was isolated from marine sponge Callyspongia elegans in Jeju Island, Republic of Korea. Strain CE80^T grew optimally at 25℃, in the range of pH 5.0-11.0 (optimum 7.0-8.0), and with 1.0-5.0% NaCl (optimum 1-3% (w/v)). Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain CE80^T belonged to the genus Labrenzia and was closely related to L. suaedae YC6927^T (98.3%), L. alexandrii DFL-11^T (96.6%), L. aggregata IAM 12614^T (96.6%) L. marina mano18^T (96.5%) and L. alba CECT 5094^T (96.2%). The major fatty acids of strain CE80^T were C_18:1 ω7c, and summed feature. The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamin, one unidentified aminolipid, one phospholipid and four unidentified lipids. The DNA G+C content of strain CE80^T was 55.9 mol%. The major respiratory quinone was Q-10. DNA-DNA relatedness between strain CE80^T and L. suaedae YC6927^T was 56.1±2.8%. On the basis of physiological and biochemical characterization and phylogenetic and chemotaxonomic analysis, strain CE80^T represents a novel species of the Labrenzia, for which the name Labrenzia callyspongiae sp. nov., is proposed. The type strain is CE80^T (=KCTC 42849^T =JCM 31309^T).

• Research in Plant Disease
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• v.16 no.1
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• pp.97-100
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• 2010

Angular leaf spot of strawberry caused by Xanthomonas fragariae occurred sporadically in the farmers open field at Sugok-myon, Jinju city, and greenhouse at Okjong-myon, Hadong-gun, Gyeongnam province, Korea. Typical symptoms of angular leaf spot initially appeared as a minute, water-soaked, rotted lesion on the lower leaf surface and the minute spots expanded to form angular leaf spots, usually delimited by small veins. The lesion was translucent under the transmitted light and dark green under reflected light. The slow growing pathogen was a gram-negative, motile bacterium with single polar flagellum, and producing yellowish orange pigment on YDC medium. It showed very strong pathogenicity on the leaves of strawberry. On the basis of biological characteristics and pathogenicity test on host plants, the bacteria was identified as a Xanthomonas fragariae. This is the first report on angular leaf spot of strawberry caused by X. fragariae in Korea.