• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1613-1621
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• 2006

Isolation and identification of algal lytic bacteria were carried out. Nine strains of algal lytic bacteria were isolated by the double-layer method using Anabaena flos-aquae as a sole nutrient. The isolate, AFK-13, showing the highest algal lytic activity was identified as Sinorhizobium kostiense based on the l6S rDNA sequence. The algal lytic experiments of the culture supernatants of AFK-13 demonstrated that the bacterial cell growth reached a maximum at 36-h culture, but the supernatant of 72-h culture exhibited the highest activity. Components among the extracellular products in the crude enzyme of the supernatant from S. kostiense AFK-13 culture were responsible for degradation of cell walls of Anabaena flos-aquae. Algal lytic assay tests of the culture supernatants suggest that the main substances for algal lytic activity could be proteinaceous. The activity of glucosidase was observed highly by polysaccharolytic analysis using the crude enzyme from S. kostiense AFK-13, whereas activities of galactosidase, mannosidase, rhamnosidase, and arabinosidase were also detected in low levels. The molecular weights (MW) of ${\alpha}-\;and\;{\beta}$-glucosidases were estimated to be approximately 50-100 kDa by the ultrafiltration method.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.745-752
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• 2007

A $\beta$-glucosidase from the algal lytic bacterium Sinorhizobium kostiense AFK-13, grown in complex media containing cellobiose, was purified to homogeneity by successive ammonium sulfate precipitation, and anion-exchange and gel-filtration chromatographies. The enzyme was shown to be a monomeric protein with an apparent molecular mass of 52 kDa and isoelectric point of approximately 5.4. It was optimally active at pH 6.0 and $40^{\circ}C$ and possessed a specific activity of 260.4 U/mg of protein against $4-nitrophenyl-\beta-D-glucopyranoside$(pNPG). A temperature-stability analysis demonstrated that the enzyme was unstable at $50^{\circ}C$ and above. The enzyme did not require divalent cations for activity, and its activity was significantly suppressed by $Hg^{+2}\;and\;Ag^+$, whereas sodium dodecyl sulfate(SDS) and Triton X-100 moderately inhibited the enzyme to under 70% of its initial activity. In an algal lytic activity analysis, the growth of cyanobacteria, such as Anabaena flos-aquae, A. cylindrica, A. macrospora, Oscillatoria sancta, and Microcystis aeruginosa, was strongly inhibited by a treatment of 20 ppm/disc or 30 ppm/disc concentration of the enzyme.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.14-19
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• 2000

Algal lytic enzyme, an extracellular enzyme, was purified from the culture filtrate of Penicillium oxalicum(HCLF-34) by ultrafiltration, gel filtration chromatography, and anion exchange chromatography. The enzyme has a molecular mass of approximately 22 kDa, an it is a monomer by renaturation SDS-PAGE. The amino acid sequences of the enzyme was revealed to be NH2-Glu-Ser-Tyr-Ser-Ser-Asn-Ala-Ala-Gly-Ala-Val-Leu-Ile---, had about 84% identity with the mature light chain of aspergillopepsin II precursor and 81% identity with the mature protein of the acid proteinase EapC precursor.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.382-390
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• 2006

Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

• Korean Journal of Ecology and Environment
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• v.36 no.2 s.103
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• pp.108-116
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• 2003

To investigate bacteria with algal Iytic activities against Anabaena cylindrica when water blooming occurs and to study enzyme profiles of alga-Iytic bacteria, various bacterial strains were isolated from surface waters and sediments in eutrophic lakes or reservoirs in Korea. Abacterial strain AK-07 was characterized and identified as Acinetobacter johnsonii based on its16S rDNA base sequence. When AK-07 was co-cultivated with A. cylindrica, bacterial cells propagated to $8\;{\times}\;10^8$ cfu $ml^{-1}$ and Iyses algal cells. However, culture filtrates of AK-07 did not exhibit algal Iytic activities. That suggesting the enzymes on the surfaces of the bacterium might be effective algal Iytic agents to cause Iyses of cells. Acinetobacter johnsonii AK-07 exhibited high degradation activities against A. cylindrica, and formed alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, glycosidases for example ${\beta}$-galatosidase, ${\beta}$-glucosidase, ${\beta}$-glucosaminidase, and ${\beta}$-xylosidase, which hydrolyzed ${\beta}$-0-glycosidic bonds, were found in cell-free extracts of A. johnsonii AK-07. Other glycosidase such as ${\alpha}$-galctosidases, ${\alpha}$-N-Ac-galctosidases, ${\alpha}$-mannosidases, and ${\alpha}$- L-fuco-sidases, which cleavage ${\alpha}$-0-glycosidic bondsare not detected. In the results, enzyme systemsof A. johnsonii AK-07 were very complex to do-grade cell walls of cyanobacteria. The polysaccharides or peptidoglycans of A. cylindrica maybe hydrolyzed and metabolized to a range of easily utilizable monosaccharides or other low molecular weight organic substances by strain AK-07 of A. johnsonii.

• Journal of Marine Bioscience and Biotechnology
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• v.10 no.1
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• pp.9-17
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• 2018

The aim of this study is to elucidate the biochemical properties of manno-oligosaccharides (MOS) hydrolyzed by extracellular enzyme of Bacillus N3. We strived to characterize the biochemical properties of MOS since N3 can effectively hydrolyzed natural polymannans such as galactomannan (GM) and konjac (glucomannan, KM), respectively. The hydrolysis of GM and KM was applied by the strain N3 in terms of reducing sugars and the highest production of reducing sugars was estimated to be about 750 mg/L and 370 mg/L respectively, which were quantified after 7 days of cultivation in the presence of both substrates. Hydrolysates derived from the hydrolysis of KM showed the significant antioxidant activity based on DPPH and ABTS radical scavenging activity with increasing of tyrosinase inhibitory activity. On the other hand, hydrolysates derived from the hydrolysis of GM showed only ABTS radical scavenging activity without showing significant changes on tyrosinase inhibitory activity. Our data suggest that those biological characteristics may be depend on the primary structure and the size of MOS, which may be useful as potent additives for diet foods.