• Journal of Life Science
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• v.29 no.2
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• pp.158-163
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• 2019

The purpose of this study was to isolate a new marine agarase-producing bacterium. Agarase can hydrolyze agar and agarose to produce agarooligosaccharides or neoagarooligosaccharides, which possess many physiological functions. Strain DH-3 was isolated from seawater collected from the coast of Yeosu at Jeollanam province, Korea. A 16S rDNA sequence analysis showed this strain to be Persicobacter sp. DH-3. Extracellular agarase was prepared from culture media of Persicobacter sp. DH-3 and used for characterization. Relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 50, 55, 70, 100, 90, and 50%, respectively. Relative activities at pH 5, 6, 7, and 8 were 75, 100, 90, and 75%, respectively. The enzyme showed maximum activity at $50^{\circ}C$ in a 20 mM Tris-HCl buffer at pH 6. This enzyme could be useful, as agar is in liquid state at $50^{\circ}C$. Agarase activities were maintained at 80% or more for 2 hr at 20, 30, and $40^{\circ}C$. Thin layer chromatography analysis suggested that Persicobacter sp. DH-3 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarohexaose and neoagarotetraose. In addition, zymogram analysis confirmed that Persicobacter sp. DH-3 produces at least three agar-degrading enzymes with molecular weights of 45, 70, and 140 kDa. Therefore, it is expected that agarases from Persicobacter sp. DH-3 could be used to produce functional neoagarooligosaccharides.

• Journal of Life Science
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• v.31 no.5
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• pp.496-501
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• 2021

In this study, the marine agar-degrading bacterium Pseudoalteromonas sp. JH-1 was isolated, and its growth and agarase properties were investigated. Seawater was collected from the offshore of the Yonggung Temple in Busan, and agar-degrading bacteria were isolated and cultured with marine agar medium. The bacterium Pseudoalteromonas sp. JH-1 was isolated through 16S rRNA gene sequencing. The extracellularly secreted enzyme was obtained from the culture broth of Pseudoalteromonas sp. JH-1 and was used to characterize its agarase. The extracellular agarase exhibited a maximum activity of 116.6 U/l at 50℃ and pH 6.0 of 20 mM Tris-HCl buffer. Relative activities were 31, 59, 94, 100, 45, and 31% at 20, 30, 40, 50, 60, and 70℃, respectively. Relative activities were 49, 85, 100, 86, 81, and 67% at pH 4, 5, 6, 7, 8, and 9, respectively. Residual activity was more than 85% after exposure at 20, 30, and 40℃ for 2 hr, and more than 82% after exposure at 50℃ for 2 hr. Zymogram analysis confirmed that Pseudoalteromonas sp. JH-1 produced at least two agarases of 55 and 97 kDa. As the products of α-agarase and β-agarase have antioxidation, antitumor, skin-whitening, macrophage activation, and prebiotic effects, further studies are needed on the agarase of Pseudoalteromonas sp. JH-1.

• Journal of Life Science
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• v.32 no.4
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• pp.285-289
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• 2022

In this study, the growth characteristics of an agar-degrading bacterium isolated from seawater samples collected from Yeongheungdo, Incheon, and the characteristics of its agarase were analyzed. The 16S rRNA gene sequence of the isolated strain was 95% similar to that of the genus Agarivorans, and thus the isolated strain was named Agarivorans sp. HY-1. When Agarivorans sp. HY-1 was cultured in a marine broth 2216 medium at 27℃ and 250 rpm, it showed maximum growth on day 1 and showed maximum enzymatic activity on day 2. A crude enzyme solution was prepared from secreted agarase in the culture medium. The extracellular agarase of the Agarivorans sp. HY-1 strain showed maximal activity at 40℃ and pH 7.0 (20 mM Tris-HCl) with 591.91 U/l. The agarase exhibited relative activities of 64, 91, 100, 97, 89, and 60% at 20, 30, 40, 50, 60, and 70℃, respectively. At pH 5, 6, 7, and 8, the relative activities were 79, 95, 100, and 55%, respectively. Furthermore, the agarase exhibited >86% residual activity at 20, 30, and 40℃ for 2 hr and >44% residual activity at 50℃ after 2 hr. A TLC analysis confirmed that Agarivorans sp. HY-1 produced α-agarase. As the degradation products of α-agarase have anticancer and antioxidant effects, Agarivorans sp. HY-1 and its agarase may well prove useful.

• Journal of Life Science
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• v.33 no.8
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• pp.680-691
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• 2023

Peak oil, climate change, and microplastics caused by the production and usage of petroleum-based plastics have threatened the sustainability of our daily life, and this has emerged as a recent global issue. To solve this global issue, the production and usage of biodegradable eco-friendly bioplastics such as polyhydroxyalkanoates (PHAs) has been suggested as an alternative. Therefore, in this review, the present status of global PHA manufacturers, the advantages of the production of PHAs using marine-origin microorganisms (with their productivity potential) and further required research and development strategies for cost-competitive production of PHAs using marine-based microorganisms were investigated. In this review, PHAs produced from marine microorganisms were found to have similar physical properties to petroleum-based plastics but with several advantages that can reduce the costs of PHA production. Those advantages include, seawater used in the medium preparation step, and osmotic-based cell lysis technology used in the separation and purification steps. However, the PHA productivities from marine microorganisms showed somewhat lower efficiencies than those from the commercial strains isolated from terrestrial environments. In order to solve the problem, further research strategies using synthetic microbiology-based technology, the development of long-term continuous culture technology, and solutions to improve PHA efficiency are required to meet future market demands for alternative bioplastics.

• KSBB Journal
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• v.24 no.4
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• pp.377-382
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• 2009

The culture condition of growing Chlorella minutissima was optimized to produce biodiesel for fed-batch cultivation. First, under heterotrophic cultivation, the optimum level of glucose was determined to be 10 g/L for 20 days. After, three cultivation conditions were operated: autotrophic, heterotrophic, and mixotrophic growth. The lipid level and the maximum cell concentration from the fed-batch heterotrophic process were 32.0 (%, v/v) and 15.0 (g-dry wt./L) in 20 L flask, respectively. In addition, since the relatively constant specific lipid production rate was observed as 0.040 (% lipid/g-dry wt./day) at the latter period of cultivation time, the fed-batch process could maintain continuous lipid production. Fed-batch process is higher than those values from the batch process. The lipids from the fed-batch process contained over 38% of $C_{18}$, known as the suitable composition for the biodiesel application. For mixotrophic and heterotrophic growth under fed-batch condition, glucose was proved to be an appropriate carbon source for a large scale outdoor cultivation. For fed-batch cultivation, the feeding rate of seawater medium containing glucose was decided to be 0.5 L/day. The mixotrophic cultivation maintained maximum cell concentration of 24 (g-dry wt./L) and the lipid level of 43 (%, w/w). The lipid composition from this process was also proved to be suitable for the biodiesel production. The fatty acids from the mixotrophic growth contains 18% of $C_{17}$ and 49% of $C_{18}$, implying It also tells that C. minutissima is a suitable resource of biodiesel. Especially, the mixotrophic cultivation with fed-batch process might be useful for the large scale cultivation for the biodiesel production.

• Korean Journal of Ichthyology
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• v.18 no.3
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• pp.202-208
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• 2006

An experiment was conducted to investigate the effects of three water temperatures (15, 20 and $25^{\circ}C$) in combination with three salinities (0, 15 and 30 psu) on the oxygen consumption rate of juvenile spotted sea bass, Lateolabrax maculatus (mean body weight $5.5{\pm}0.3g$). The oxygen consumption rates of L. maculatus were measured in triplicate for 24 hours using a continuous flow-through respirometer. Water temperature resulted in significant differences in the mean oxygen consumption rate of L. maculatus (p<0.001), but salinity and combinations of salinity and water temperature did not have (p>0.05). The oxygen consumption increased with increasing water temperatures in all experimental salinity regimes (p<001). Mean oxygen consumption rates at 15, 20 and $25^{\circ}C$ ranged 328.8~342.3, 433.9~441.0 and 651.5~659.9 mg $O_2\;kg^{-1}\;h^{-1}$, respectively. $Q_{10}$ values did not vary with salinity, bud varied with water temperature. $Q_{10}$ values ranged 1.63~1.75 between 15 and $20^{\circ}C$, 2.24~2.26 between 20 and $25^{\circ}C$, and 1.92~1.98 over the full temperature range. The energy loss by metabolic cost increased with increasing water temperatures in all experimental salinity regimes (p<0.001) Mean energy loss rates at 15, 20 and $25^{\circ}C$ ranged 224.6~233.8, 296.3~301.2 and $444.9{\sim}450.7kJ\;kg^{-1}\;d^{-1}$, respectively. These data suggest that the culture of juvenile spotted sea bass is possible without energy loss by salinity difference in freshwater as well as seawater after salinity acclimation. Thus, this result has an application for culture management and bioenergetic model for growth of this species.

• Journal of Life Science
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• v.28 no.9
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• pp.1056-1061
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• 2018

This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.2
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• pp.91-97
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• 2001

The effects of sudden changes of water temperature (WT) on the stress response and physiological change of the cultured olive flounder in large (FL) and small (FS) size, and fat cod (FC) were examined by manipulating WT (2 types) in a flow through seawater culture system with 6 tanks (water vol. 270 L/tank). The WT was decreased from $20^{\circ}C$ to $10^{\circ}C$ within 5 hours ($2^{\circ}C/hour$) and maintained at $10^{\circ}C$ for 21 hours (Exp. I), and it was raised from $20^{\circ}C$ to $30^{\circ}C$ within 5 hours and maintained at $30^{\circ}C$ for 21 hours (Exp.II). In Exp. I, the levels of blood hematocrit at 5 hours ($10^{\circ}C$) in FS was significantly decreased from $13.5\pm2.0\%\;to\;11.3\pm2.3\%$, but FC at 2.5 hours ($15^{\circ}C$) ($19.0\pm0.3\%\;to\;23.2\pm3.8\%$) was increased, The blood hemoglobin concentration of all fish in Exp, II was significantly increased until 8 hours after raising WT from $20^{\circ}C$ to $30^{\circ}C$. In Exp. I and II , the levels of plasma cortisol in FL, FS and FC was changed from $5.2\pm8.5ng/mL,\;4.4\pm4.5ng/mL\;and\;2.7\pm0.4ng/mL$, respectively, before sudden drop and rise of WT. The levels of plasma cortisol of in FL ($164.0\pm53.1ng/mL$) and FC ($207.9\pm25.4ng/mL$) were significantly increased by the lowering WT sharply during whole experiment. The FL ($12.6\pm2.0ng/mL$) and FS ($4.0\pm3.9ng/mL$) showed no significant differences in cortisol level according to sudden rise of WT (5 hours). But it in FC ($44.7\pm18.2ng/mL$) was increased. In Exp. I, the plasma glucose levels of all fish groups were decreased after 5 hours ($10^{\circ}C$), The plasma lactic acid concentration of FL and FS showed no significant differences until 5 hours after raising WT from $20^{\circ}C$ to $30^{\circ}C$, But it in FC was significantly increased with WT raise.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.5
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• pp.465-472
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• 2001

The effects of water level reduction in rearing tank and fish transference during fish selection process on the stress response (hematological factors, cortisol, glucose, lactic acid and osmolality) of tank-reared olive flounder Paralichthys olivaceus of large (FL), small (FS) and Japanese croaker, Nibea japonica (JC) were examined in running seawater culture system. The water level of rearing unit was lowered from 33 cm to 8 cm in the course of 2 minutes in the water level reduction experiment. The fish were removed from rearing tank (12 ton) to 450 L tank in 30 seconds after capture in the fish transference experiment, In water level reduction, the hematocrit of FL was significantly increased from $14.6\%$ at beginning to $23.5\%$ after 10 hours, However, it decreased to the value of beginning after 46 hours. Plasma cortisol concentration of FL was the highest concentration (13.7 ng/mL) after 22 hours, but it decreased to 4.0 ng/mL at the end of experiment. Cortisol concentration of FS did not show any significant difference during the experiment. The cortisol concentration of JC were significantly higher at 4 hours (282.3 ng/mL) and 22 hours (350.5 ng/mL), Glucose concentration of JC was the highest (138.0 mg/dL) at 22 hours. Lactic acid concentration was not different between experimental groups. In the fish transference experiment, red blood cell of FL was increased from $1.9\times10^6\;cell/{\mu}\;L\;to\;4.2\times10^6\;cell/{\mu}L$ in 24 hours. Blood hemoglobin of JC were significantly elevated in 24 hours. At 1 hour after transference, plasma cortisol concentrations in both fish species were increased to 95.3 ng/mL in FL and 175.5 ng/mL in JC. Glucose concentration of JC was increased to 132.5 mg/dL at 1 hour, 129.5 mg/dL at 3 hours after transference.

• The Korean Journal of Malacology
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• v.17 no.1
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• pp.45-55
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• 2001

Gametogenes, reproductive cycle, first sexual maturity(biological minimum size), sex ratio and larval development of the marsh clam Corbicula japonica were investigated monthly by histological observations. Samples were collected in brackish water of Gokgang stream, Kyungsangbuk-Do, Korea, from August 1997 to July 1998. Sexuality of Corbicula japonica is dioecious and the species are an oviparous clam. The gonads are irregularly arranged from the sub-region of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary is composed of a number of ovarian sac which are branched arborescent. Oogonia actively proliferate along the germinal epithelium of ovarian sac, in which young oocytes are growing. The testis is composed of a number of testicular tubules, and the epithelium of the tubule has function of germinal epithelium, along which spermatogonia actively proliferate. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed between developing oocytes and spermatocytes in the early developmental stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells are considered to be related to the growing of the oocytes and spermatocytes. The spawning period is from July to September, and the main spawning occur between July and August when seawater temperatures reach above 22$^{\circ}C$. The reproductive cycle of this species can be divided into five successive stages; early active (February to April), late active (May to July), ripe (June to September), partially spawned (July to September), degenerative (September to October) and resting stage (October to February). Percentages of first sexual maturity of female and male clams ranging in length from 10 mm to 12 mm are over 50% and 100% for clams over 16.0 mm in shell length. Fertilized eggs or Corbicula japonica were 80-90 ${\mu}{\textrm}{m}$ in diameter. In the early embryonic development of C. japonica, the appearance of polar body, trochophore and D-shaped veliger were observed around 40 min., 27 hours and 4 days after spawning, respectively, at a water temperature of 26.5-28.$0^{\circ}C$. The size of larvae of early umbo stage was about 185-210 ${\mu}{\textrm}{m}$ in shell length, 160-180 ${\mu}{\textrm}{m}$ in shell height around 7 days after fertilization. The correlation of relative growth between the culture day (D) and shell length (SL) was expressed by the following simple formula from D-shaped veliger to metamorphosing stage; SL = 13.300D + 209.36($r^2$= 0.9078).