• Microbiology and Biotechnology Letters
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• v.40 no.3
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• pp.243-249
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• 2012

This study was conducted to screen an alginate-degrading microorganism and to investigate the characteristics of the alginate-degrading activity of its crude enzyme. A marine bacterium which produces extracellular alginate-degrading enzymes was isolated from the brown alga Sargassum thunbergii. 16S rRNA sequence analysis and physiological profiling resulted in the bacterium's identification as a Vibrio crassostreae strain, named Vibrio crassostreae PKA 1002. Its optimal culture conditions for growth were pH 9, 2% NaCl, $30^{\circ}C$ and a 24 hr incubation time. The optimal conditions for the alginate degrading ability of the crude enzyme produced by V. crassostreae PKA 1002 were pH 9, $30^{\circ}C$, a 48 hr incubation time and 8% alginic acid. The alginate degrading crude enzyme produced 3.035 g of reducing sugar per liter in 4% (w/v) alginate over 1 hr.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.79-84
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• 2020

This study investigated the characterization and functionality of Undaria pinnatifida root (UPT) extracts, degraded using a crude enzyme from Shewanella oneidensis PKA1008. To obtain the optimum degrading conditions, the UPT was mixed with alginate degrading enzymes from S. oneidensis PKA 1008 and was incubated at 30℃ for 0, 3, 6, 12, 24, and 48 h. The alginate degrading ability of these enzymes was then evaluated by measuring the reducing sugar, viscosity, pH and chromaticity. Enzymatic extract at 24 h revealed the highest alginate degrading ability and the lowest pH value. As the incubation time increased, the lightness (L ^⁎) also decreased and was measured at its lowest value, 39.84, at 12 hours. The redness and yellowness increased gradually to 10.27 at 6 h and to 63.95 at 3 h, respectively. Moreover, the alginate oligosaccharides exhibited significant anti-inflammatory activity. These results indicate that a crude enzyme from S. oneidensis PKA 1008 can be used to enhance the polysaccharide degradation of UPT and the alginate oligosaccharides may also enhance the anti-inflammatory effect.

• Microbiology and Biotechnology Letters
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• v.41 no.3
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• pp.372-378
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• 2013

An alginate-degrading bacterium, identified as Shewanella oneidensis PKA 1008 by 16S ribosomal RNA sequence analysis, was isolated from the green alga Ulva pertusa. Optimal conditions for the alginate-degrading ability of its crude enzyme were then determined. The optimal culture conditions for the growth of S. oneidensis PKA 1008 were pH 9, 2% NaCl, $30^{\circ}C$, and 24 hours incubation time. The crude enzyme produced by S. oneidensis PKA 1008 showed the highest alginate-degrading activity at pH 9, $30^{\circ}C$ and produced 1.001 g of reducing sugar per liter in 3.5% (w/v) sodium alginate for 1 hour.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.3
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• pp.434-440
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• 2013

This study was conducted to screen the characteristics and alginate-degrading activity of marine bacterium isolated from brown seaweed (Sargassum thunbergii). The results of 16S ribosomal RNA sequence analysis the strain the genus Vibrio and the strain was subsequently named Vibrio sp. PKA 1003. The optimum culture conditions for the growth of Vibrio sp. PKA 1003 were at pH 7, 3% NaCl, $25^{\circ}C$, and 6% alginic acid, with a 48-hour incubation time. A crude enzyme preparation from Vibrio sp. PKA 1003, showed its highest levels of alginate-degradation activity when cultured at pH 9, $30^{\circ}C$, and 6% alginic acid, with a 63-hour incubation time. Thin layer chromatography analyses confirmed that the crude enzyme released monomers or oligomers from sodium alginate, and results from trypsin treatment showed that the alginate degrading activity depends on this enzyme produced by Vibrio sp. PKA 1003. These results suggest that Vibrio sp. PKA 1003 and its alginate-degrading crude enzyme is useful for the production of alginate oligosaccharides.

• Journal of Microbiology
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• v.44 no.3
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• pp.354-359
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• 2006

This study examined the capacity of immobilized bacteria to degrade petroleum hydrocarbons. A mixture of hydrocarbon-degrading bacterial strains was immobilized in alginate and incubated in crude oil-contaminated artificial seawater (ASW). Analysis of hydrocarbon residues following a 30-day incubation period demonstrated that the biodegradation capacity of the microorganisms was not compromised by the immobilization. Removal of n-alkanes was similar in immobilized cells and control cells. To test reusability, the immobilized bacteria were incubated for sequential increments of 30 days. No decline in biodegradation capacity of the immobilized consortium of bacterial cells was noted over its repeated use. We conclude that immobilized hydrocarbon-degrading bacteria represent a promising application in the bioremediation of hydrocarbon-contaminated areas.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.56 no.2
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• pp.204-211
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• 2023

This study investigated the physicochemical properties and palatability-enhancing effects of the alcohol precipitate, in the enzymatic extracts of Sargassum confusum C. Agardh (SC), obsained using the crude enzyme of Shewanella oneidensis PKA 1008. We analyzed the oligosaccharides recovered from the alcohol precipitate using a thin-layer chromatography for SC-degrading extracts, pH, color, reducing sugar, and viscosity. Thin-layer chromatography showed that after treating with the crude enzyme for 60 h, the polysaccharides were degraded into tetramers, dimers, and trimers and pH increased in the alcohol precipitate (EtOH Sedi). In terms of color, the redness and yellowness of alcohol precipitate/supernatant (EtOH Sedi+Super) and the brightness of EtOH Sedi were the highest among enzyme treated for 0 h and 60 h, EtOH Sedi, and EtOH Sedi+Super. In the reducing sugar analysis, EtOH Sedi showed the lowest value of 13.63 ㎍/mL, and the lowest viscosity of 1.13. In terms of the sensory evaluation, EtOH Sedi+Super showed the highest value with respect to the overall preference. These results suggest that the crude enzyme of S. oneidensis PKA 1008 is effective at degrading polysaccharides, and its recovery increases the palatability of the alcohol precipitate.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.364-373
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• 2015

As an effort to utilize alginate, 103 bacterial isolates that were positive for the alginate lyase activity were isolated from various clams and seawater samples collected in Incheon coastal area. Among them, 3 strains (M1-2-1, M6-1, and C8-15) were finally selected for further analysis based on their activities at higher levels than others. These isolates were all Gram-negative and rod shaped halophilic bacteria with motility. According to their physiological and biochemical properties as well as DNA sequence of their 16S rRNA genes, M1-2-1 and M6-1 were identified as a member of genus Pseudoalteromonas and C8-15 belonged to genus Vibrio. They exhibited the alginate degrading activity at the maximal level when they were cultured in APY broth for 6-8 h at $25^{\circ}C$. Both their growth and the enzyme activity were greatly enhanced when NaCl was added to the growth medium. The crude alginate lyases from the supernatants of the bacterial cultures showed the highest activity at $45^{\circ}C$ and pH 7.0-8.0. M1-2-1 and M6-1 produced 2.723 and 1.976 g/L of reducing sugar from alginate, respectively, suggesting that they have potential for commercial application.

• Microbiology and Biotechnology Letters
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• v.43 no.2
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• pp.105-111
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• 2015

An alginate degrading enzyme from the Vibrio crassostreae PKA 1002 strain was used to hydrolyze the water extract of Sargassum thunbergii. To obtain the optimum degrading conditions for the S. thunbergii water extract, the mixture of the water extract and enzyme was incubated at 30℃ for 0, 3, 6, 12, and 24 h, and its alginate degrading ability was measured by reducing sugar and viscosity. A temperature of 30℃ for a period of 6 h was found to be the optimal condition for the enhancement of the alginate’s degrading ability. The pH of the enzymatic hydrolysate was not significantly different from that of the water extract. Overall lightness decreased, but redness and yellowness increased after enzymatic hydrolysis. Total phenolic compounds did not differ between the water extract and the enzymatic hydrolysate. DPPH radical scavenging activity and the reducing power of the enzymatic hydrolysate were lower than those of the water extract. However, the chelating effect of the enzymatic hydrolysate (80.08% at 5 mg/ml) was higher than that of the water extract (62.29%). These results indicate that the enzymatic hydrolysate possesses an anti-oxidant activity by way of the action of the chelating effect.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.8
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• pp.919-928
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• 2017

The immuno-enhancing effects of alginate oligosaccharides from Sargassum coreanum were investigated. The alginate oligosaccharides were produced by an alginate-degrading enzyme from S. oneidensis PKA 1008. The degraded alginate oligosaccharides were visualized by thin-layer chromatography developed using a solvent system of 1-butanol/methanol/water, 4:1:2 (v/v/v). Alginate was degraded into dimmers at 60 h. As a result, the levels of Th1 cytokine [interferon $(IFN)-{\gamma}$ and interleukin (IL)-2] and Th2 cytokine (IL-6 and IL-10) increased with increasing incubation time compared to the control in vitro. Enzymatic extract treatment promoted proliferation of splenocytes at concentrations of 100 and 200 mg/kg at 24 h in vivo. Secretion of $IFN-{\gamma}$ and IL-2 significantly increased in a dose-dependent manner at 24 h as well as induced higher production of IgG2a in serum. Natural killer cell activity was measured and tended to increase. In addition, complete blood cell counts increased in a dose-dependent manner. These results indicate that alginate oligosaccharides produced by crude enzyme from S. oneidensis PKA 1008 may have significant immune activities.