• Journal of Life Science
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• v.26 no.4
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• pp.453-459
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• 2016

Agarases are classified into α-agarase and β-agarase that produce agarooligosaccharides and neoagarooligosaccharides, respectively. Neoagarooligosaccharides have whitening effect of skin, delay of starch degradation, and inhibition of bacterial growth etc. Hence, the object of this study was to isolate a novel agarase producing marine bacterium and characterization of its β-agarase. A novel agar-degrading bacterium was isolated from seashore of Namhae at Gyeongnamprovine, Korea and purely cultured with Marine agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-4 after 16S rRNA gene sequencing. The enzymatic sample was obtained from culture media of Simiduia sp. SH-4. Enzymatic activity was highly increased from 20(30% relative activity) to 30℃ (100%) and decreased from 30 to 40℃(75%) and so more. Relative activity was 100% at pH 6 while those were about 91% and 59% at pH 5.0 and 7.0, respectively, meaning the enzyme possesses narrow optimal pH range. Hence, the enzyme exhibited the maximal activity with 120.4 units/l at pH 6.0 and 30℃ in 20 mM Tris-HCl buffer. Thin layer chromatography (TLC) analysis showed that Simiduia sp. SH-4 produces β-agarase, which hydrolyze agarose to produce biofunctional neoagarooligosaccharides such as neoagarotetraose and neoagarobiose. Hence, broad applications would be possible using Simiduia sp. SH-4 and its enzyme in the food industry, cosmetics and medical fields.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.104-110
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• 2004

To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.

• Food Science of Animal Resources
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• v.31 no.6
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• pp.952-959
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• 2011

A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to $100^{\circ}C$ for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and $C_{18}$ column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was $NH_2$-ILLEELNV.

• The Korean Journal of Mycology
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• v.49 no.2
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• pp.199-209
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• 2021

The present study was performed to improve the technique used for fermenting the mushroom growth medium. Taxonomic analysis of 16S rDNA sequence from the predominant Bacillus strain CY-24 isolated during the fermentation phase of the rice straw medium identified it as Bacillus licheniformis. In addition, the growth environment of B. licheniformis was also examined in this study, which revealed the optimal growth temperature and pH to be 30 ℃ and 6.0, respectively. This study also revealed that carboxymethyl cellulase (CMCase) and polygalacturonase (PGase) enzymes isolated from B. licheniformis achieved their maximal activities at 50 ℃ and 60 ℃ respectively. Furthermore, the study confirmed that the two enzymes, i.e., CMCase and PGase in B. licheniformis are stable at temperatures above 60 ℃. The present study thus demonstrates that B. licheniformis CY-24 possesses excellent enzymatic properties. It also reveals that the action of enzymes during the production of growth mediums used for the cultivation of mushrooms is closely associated with the promotion of fermentation and softening of the rice straw. Overall, this study provides elementary information regarding the role of B. licheniformis enzymes during growth medium fermentation for Agaricus bisporus cultivation.

• Korean Journal of Microbiology
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• v.39 no.1
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• pp.16-20
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• 2003

A phylogenetic analysis of bacterial populations inhabiting soil derived from serpentine was conducted. The samples were collected from adjacent metamorphic rocks and serpentinite soil at Kwangcheon. The pH of the serpentine areas ranged from 8.5 to 9.2. The number of bacteria on the DAL medium which was diluted with $10^{-2}$ of AL medium was 10~100 fold higher than that from the full strength of AL medium, and which indicates that oligotrophs are distributed in the serpentinite soil. Of a total of 76 isolates, 42 isolates were oligotrophic bacteria, which grew only on the DAL medium. Based on a phylogenetic analysis using 16S rDNA sequences, these isolates are found to fall within five major phylogenetic groups: proteobacteria $\alpha$-subdivision (3 strains), $\alpha$-subdivision (7 strains), $\gamma$-subdivision (2 trains); high G+C gram-positive bacteria (19 strains); low G+C grampositive bacteria (14 strains). Bacteria of the genus Streptomyces (high G+C division) and Bacillus (low G+C division) have been considered to form a numerically important fraction of serpentinite soil. Oligotrophic strains categorized as Afipia ($\alpha$-subdivision), Ralstonia, Variovorax ($\beta$-subdivision), Pseudomonas ($\gamma$ -subdivision), Arthrobacter (high G+C division), and Streptomyces (low G+C division).

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.156-162
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• 2016

In this study, we isolated a new agar-degrading marine bacterium and characterized its agarase. An agardegrading marine bacterium SH-1 was isolated from seawater, collected from the seashore of Namhae in Gyeongnam province, Korea, and cultured in marine agar 2216 media. It was identified as Maribacter. sp. SH-1 by phylogenetic analyses, based on 16S rRNA gene sequence. The extracellular agarase was extracted from culture media of Maribacter sp. SH-1 and characterized. Its relative activities were 56, 62, 94, 100, and 8% at 20, 30, 40, 50, and 60℃, respectively, whereas 15, 100, 60, and 21% relative activities were observed at pH 5, 6, 7, and 8, respectively. Its extracellular agarase exhibited maximum activity (231 units/l) at pH 6.0 and 50℃, in 20 mM Tris-HCl buffer. Therefore, this agarase would be applicable as it showed the maximum activity at the temperature at which the agar is in a sol state. Furthermore, the agarase activities remained over 90% at 20, 30, and 40℃ after 0.5 h exposure at these temperatures. Thin layer chromatography analysis suggested that Maribacter sp. SH-1 produces extracellular β-agarase, as it hydrolyzes agarose to produce neoagarooligosaccharides, such as neoagarohexaose (34.8%), neoagarotetraose (52.2%), and neoagarobiose (13.0%). Maribacter sp. SH-1 and its β-agarase would be useful for the production of neoagarooligosaccharides, which shows functional properties, like skin moisturizing, skin whitening, inhibition of bacterial growth, and delay in starch degradation.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.919-923
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• 2004

The sk10 isolated from kimchi was identified as W. kimchii on the basis of l6s-rDNA sequencing. Studies were made to analyze the metabolic flux shift of the sk10 on glucose under aerobic growth conditions. The sk10 produced 38.2 mM acetate, 16.3 mM ethanol, and 33.2 mM lactate under aerobic conditions, but 2.4 mM acetate, 48.0 mM ethanol, and 44.1 mM lactate under anaerobic conditions. The NADH peroxidase (NADH-dependent hydrogen peroxidase) activity of sk10 grown under aerobic conditions was 11 times higher than that under anaerobic conditions. Under the low ratio of $NADH/NAD^+$, the metabolic flux toward lactate and ethanol was shifted to the flux through acetate kinase without NADH oxidation. The kinds of enzymes and metabolites of sk10 were close to those in the pathway of Leuconostoc sp., but the metabolites produced under aerobic growth conditions were different from those of Leuconostoc sp. The stoichiometric balance calculated using the concentrations of metabolites and substrate was about 97%, coincident with the theoretical values under both aerobic and anaerobic conditions. From these results, it was concluded that the metabolic flux of W. kimchii sk10 was partially shifted from lactate and ethanol to acetate under aerobic conditions only.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.673-679
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• 2004

A microbial cryoprotectant formulation using a W/O/W multiple emulsion system was developed. The psychrotolerant microorganism, B4, isolated from soil in South Korea, was observed by the drop freezing method, in which the microorganism sample inhibited ice nucleation activity. The antifreeze activity was eliminated when the microorganism sample was treated with protease, indicating that the antifreeze activity was due to the presence of antifreeze protein. The result of the l6S rDNA sequencing indicated the B4 strain was most closely related to a species of the genus Bacillus. Culture broth of B4 strain (Bacillus sp.) and rapeseed oil containing 1 % polyglycerine polyricinolate (PGPR) were used as core and wall material, respectively. The most stable W/O emulsion was prepared at a core/oil ratio of 1:2. The highest W/O/W emulsion stability was achieved when the primary emulsion to external aqueous phase containing 0.5% caster oil polyoxyethylene ether $(COG25^{TM})$ ratio was 1:1. Microcrystalline cellulose showed better W/O/W emulsion stability than other polymer types. The viability of cells in a W/O/W emulsion was higher than free cells during storage at $37^\circ{C}$. An acidic pH and UV exposure decreased the viability of free cells, but cells in W/O/W emulsion were more stable under these conditions.

• KSBB Journal
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• v.25 no.6
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• pp.565-571
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• 2010

To develop thermostable ethanol fermentative yeast strain for lignocellulosic simultaneous saccharification and fermentation, high ethanol producing yeast, Saccharomyces cerevisiae CHY1012 and thermostable yeast, Kluyveromyces marxianus CHY1703 were fused by protoplast fusion. The thermostable fusant, CHY1612 was identified as a Kluyveromyces marxianus by phenotypic and physiological characteristics, as well as molecular analysis based on the D1/D2 domains of the large subunit (26S) rDNA gene and the internally transcribed spacer (ITS) 1 + 2 regions. For lignocellulosic ethanol production, AFEX pretreated barley straw at $150^{\circ}C$ for 90 min was used in a simultaneous saccharification and fermentation (SSF) process using thermotolerant CHY1612. The SSF from 16% pretreated barley straw at $43^{\circ}C$ gave a saccharification ratio of 90.5%, a final ethanol concentration of 38.5 g/L, and a theoretical yield of 91.2%. These results show that K. marxianus CHY1612 has potential for lignocellulosic ethanol production through simultaneous saccharification and fermentation with further development of process.

• Environmental Engineering Research
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• v.23 no.3
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• pp.258-264
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• 2018

The objectives of this research were to 1) isolate and identify thermophilic bacteria for food waste treatment; 2) investigate the capability of food waste treatment using Bacillus species; and 3) develop air-dried Bacillus starters for food waste treatment. Five Bacillus species were isolated from food wastes and identified as Bacillus licheniformis (B. licheniformis) G1, Bacillus circulans C2, Bacillus subtilis (B. subtilis) E1, Bacillus vanillea F1, and Bacillus atrophaeus G2 based on 16S rDNA sequencing. Each identified Bacillus and the mixture of Bacillus species were cultivated in the standard food waste at $45^{\circ}C$ for 8 d. Changes in cell count, solid contents, and pH of the food waste were monitored during cultivation. Air-dried Bacillus cell powders were prepared using wheat flour and lactomil as excipients, and the cell count and survival rate were determined. The cell count of B. licheniformis G1 exhibited the highest number among the tested Bacillus (${\sim}10^8CFU/mL$). The greatest reduction in solid contents of food waste was achieved by B. subtilis E1 (22.6%). The mixture of B. licheniformis G1 and B. subtilis E1 exhibited a synergistic effect on the reduction of solid contents. Lactomil was determined as better excipient than wheat flour based on the greatest survival rate of 95%.