• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2003.10a
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• pp.34-63
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• 2003

Occupational and environmental exposure to manganese continue to represent a realistic public health problem in both developed and developing countries. Increased utility of MMT as a replacement for lead in gasoline creates a new source of environmental exposure to manganese. It is, therefore, imperative that further attention be directed at molecular neurotoxicology of manganese. A Need for a more complete understanding of manganese functions both in health and disease, and for a better defined role of manganese in iron metabolism is well substantiated. The in-depth studies in this area should provide novel information on the potential public health risk associated with manganese exposure. It will also explore novel mechanism(s) of manganese-induced neurotoxicity from the angle of Mn-Fe interaction at both systemic and cellular levels. More importantly, the result of these studies will offer clues to the etiology of IPD and its associated abnormal iron and energy metabolism. To achieve these goals, however, a number of outstanding questions remain to be resolved. First, one must understand what species of manganese in the biological matrices plays critical role in the induction of neurotoxicity, Mn(II) or Mn(III)? In our own studies with aconitase, Cpx-I, and Cpx-II, manganese was added to the buffers as the divalent salt, i.e., $MnCl_2$. While it is quite reasonable to suggest that the effect on aconitase and/or Cpx-I activites was associated with the divalent species of manganese, the experimental design does not preclude the possibility that a manganese species of higher oxidation state, such as Mn(III), is required for the induction of these effects. The ionic radius of Mn(III) is 65 ppm, which is similar to the ionic size to Fe(III) (65 ppm at the high spin state) in aconitase (Nieboer and Fletcher, 1996; Sneed et al., 1953). Thus it is plausible that the higher oxidation state of manganese optimally fits into the geometric space of aconitase, serving as the active species in this enzymatic reaction. In the current literature, most of the studies on manganese toxicity have used Mn(II) as $MnCl_2$ rather than Mn(III). The obvious advantage of Mn(II) is its good water solubility, which allows effortless preparation in either in vivo or in vitro investigation, whereas almost all of the Mn(III) salt products on the comparison between two valent manganese species nearly infeasible. Thus a more intimate collaboration with physiochemists to develop a better way to study Mn(III) species in biological matrices is pressingly needed. Second, In spite of the special affinity of manganese for mitochondria and its similar chemical properties to iron, there is a sound reason to postulate that manganese may act as an iron surrogate in certain iron-requiring enzymes. It is, therefore, imperative to design the physiochemical studies to determine whether manganese can indeed exchange with iron in proteins, and to understand how manganese interacts with tertiary structure of proteins. The studies on binding properties (such as affinity constant, dissociation parameter, etc.) of manganese and iron to key enzymes associated with iron and energy regulation would add additional information to our knowledge of Mn-Fe neurotoxicity. Third, manganese exposure, either in vivo or in vitro, promotes cellular overload of iron. It is still unclear, however, how exactly manganese interacts with cellular iron regulatory processes and what is the mechanism underlying this cellular iron overload. As discussed above, the binding of IRP-I to TfR mRNA leads to the expression of TfR, thereby increasing cellular iron uptake. The sequence encoding TfR mRNA, in particular IRE fragments, has been well-documented in literature. It is therefore possible to use molecular technique to elaborate whether manganese cytotoxicity influences the mRNA expression of iron regulatory proteins and how manganese exposure alters the binding activity of IPRs to TfR mRNA. Finally, the current manganese investigation has largely focused on the issues ranging from disposition/toxicity study to the characterization of clinical symptoms. Much less has been done regarding the risk assessment of environmenta/occupational exposure. One of the unsolved, pressing puzzles is the lack of reliable biomarker(s) for manganese-induced neurologic lesions in long-term, low-level exposure situation. Lack of such a diagnostic means renders it impossible to assess the human health risk and long-term social impact associated with potentially elevated manganese in environment. The biochemical interaction between manganese and iron, particularly the ensuing subtle changes of certain relevant proteins, provides the opportunity to identify and develop such a specific biomarker for manganese-induced neuronal damage. By learning the molecular mechanism of cytotoxicity, one will be able to find a better way for prediction and treatment of manganese-initiated neurodegenerative diseases.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.24 no.5
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• pp.273-288
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• 1991

To elucidate the physiological and biochemical differences between chondrichthyes and osteichthyes, the properties of the specific digestive enzymes in cat-shark, Cephaloscyllium umbratile, and mackerel, Scomber japonicus, were studied. Homogenous trypsin proved through the disc-electrophoresis, SDS-PAG electrophoresis and gel filtration was obtained from the pancreas of cat-shark by $50-70\%$ saturated ammonium sulphate fractionation, DEAE-Sephadex A-50 column chromatography, benzamidine-Sepharose 6B affinity chromatography and Sephadex G-75-120 gel filtration. Two types of trypsins were also obtained from the pyloric caeca of mackerel by $30-70\%$ saturated ammonium sulphate fractionation and the slightly modified procedure from the method adopted in the purification of cat-shark trypsin. The two trypsins, designated trypsin A and B, were proved their homogeneity by disc- and SDS-PAG electrophoresis and gel filtration. The molecular weights of the trypsins were estimated to be 31,700 for cat-shark trypsin, 30,000 for mackerel trypsin A and 29,000 for mackerel trypsin B by SDS-PAG electrophoresis, but those were estimated to be 21,500 for cat-shark trypsin, 23,700 for mackerel trypsin A and 21,500 for mackerel trypsin B by gel filtration. The trypsins exhibited their optimum conditions at pH 9.0 and on temperature ranged from $45^{\circ}C\;to\;50^{\circ}C$ for cat-shark, and at pH 8.0 and a temperature of $50^{\circ}C$ for mackerel trypsin A and B, respectively. The cat-shark trypsin was stable at pH 10.0 and the temperature below $10^{\circ}C$, whereas the mackerel trypsin A and B, were stable in the range over pH 7.0 to pH 9.0 below $10^{\circ}C$ and at pH 8.0 below $35^{\circ}C$, respectively. The mackerel trypsins were severely inhibited by some heavy metal ions such as $Ag^{2+},\;Cu^{2+}\;and\;Hg^{2+}$ compared to cat-shark trypsin. All of the enzymes were also inhibited by antipain, leupeptin, TLCK(tosyllysine chloromethyl ketone) and SBTI(soybean trypsin inhibitor) remarkably. The inhibitory effects of PMSF(phenylmethane sulphonylfluoride), DFP(diisopropyl fluorophosphate) and benzamidine were indicated that these enzymes belong to serine-proteases.

• Food Science and Preservation
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• v.23 no.7
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• pp.1050-1057
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• 2016

This study was carried out to investigate the effectiveness of Leuconostoc mesenteroides isolated from octopus baechu kimchi as a potential starter for seafood kimchi. L. mesenteroides is lactic acid bacterium currently used as a starter for kimchi production. We selected the most effective L. mesenteroides strain from the 7 strains isolated from octopus baechu kimchi and, based on biochemical properties and 16S rRNA sequencing, identified the selected strain as L. mesenteroides SK-1. The SK-1 strain exhibited acid-tolerance, good survival capacity, and excellent dextran productivity. We investigated the effects the SK-1 of starter on seafood kimchi fermentation. Octopus baechu kimchi was fermented with L. mesenteroides SK-1 at $4^{\circ}C$ for 35 d. The decrease in pH and increase in acidity in octopus baechu kimchi fermented with the SK-1 starter occurred more quickly than that in the control kimchi indicating that. Octopus baechu kimchi with SK-1 starter has a relatively slow rate of increase in lactic acid production. As a result, octopus baechu kimchi prepared with L. mesenteroides SK-1 can be maintained at a suitable ripening degree over an extended period of time compared to that of the control kimchi, Moreover, the octopus baechu kimchi started with L. mesenteroides SK-1 has excellent sensory properties, including a refreshing taste, and a weak sour odor.

• Journal of Sericultural and Entomological Science
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• v.23 no.1
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• pp.1-31
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• 1981

Rootability of the hardwood cuttings of mulberry was related not only histological characteristics but dependent on biochemical properties. In this connection, the characteristics of the hardwood cuttings were histologically observed and the growth substances produced by the cuttings were also identified by means of mung bean bioassay. Amino acid, carbohydrate, nucleic acid contents, and the C/N ratio were also analysed. The results are summarized as follows. 1. There were differences in rootability of cuttings between mulberry species and varieties Among the three mulberry species tested, Morus Lhou Koidz. showed the highest rootability while M. bombycis showed the lowest one. In varietal differences in rootability, it was shown that the varieties could be grouped according to rootability: high varieties(above 80%), medium(41~79%), and low(below 40%). The higher varieties were Kemmochi, Nakamaki, Kosen, and Wusuba roso. 2. The histological characteristic of the hardwood cuttings most closely related to rootability was cell layer arrangement in the sclerenchyma tissue. The lower rootability varieties developed two or three overlapping cell layers in the bark tissue and in the higher rootability varieties they were scattered over the primary cortex. 3. In the higher rootability varieties, there was a positive correlation between the development of root primodia and rootability of the hardwood cuttings. It was also shown that there was a close relationship between the size of primodia and the surface area of the lenticel with rootability of the cuttings. 4. Effect of growth substances extracted from the hardwood cuttings were determined by mung bean bioassay. The higher rootability varieties usually showed higher activities of the growth substances, in contrast the lower rootability varieties showed higher activities of the inhibitory substances. 5. It was evident that the substance separated by paper chromatography was identified as indole acetic acid with $R_f$ value ranging from 0.3 to 0.5. The other substances detected at a $R_f$ value ranging from 0.8 to 1.0 and origin to 0.1 were also responsible for rooting. 6. There exists a quantitatively different distribution of growth substances in a synergistic system in the tissues of cuttings, and the balance between growth and inhibitory substances gives rise to the development of rooting. Particularly, no descent of the substances from winter buds resulted in no rooting of cuttings but these substances were produced a week after planting in a warm environment. 7. It was shown that there were positive correlations between carbohydrate ($r=0.72^*$) and total sugar ($r=0.67^*$) and rootability, respectively, but there were negative correlations between reducing sugars ($r=-0.75^*$) and rootability. 8. High C/N ratio gave rise to high rootability($r=0.67^*$). The latter therefore depended on high amount of carbohydrate rather than nitrogen in the cuttings. 9. The content of RNA and DNA in the cuttings was not changed for upto two weeks after the cuttings were planted. Then an increase in RNA content took place in only the high rootability varieties. 10. There were quantitative and qualitative differences in the compositions of the amino acids between the high rootability varieties and the low rootability varieties. More aspartic acid and cystine were found in the higher rootability varieties than in the low rootability varieties.

• Journal of Life Science
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• v.18 no.2
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• pp.264-272
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• 2008

The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Acanthogobius hasta were characterized by biochemical, immunochemical and kinetic methods. The activities of LDH in skeletal muscle and eye tissues were 65.30 and 53.25 units, but LDH activities in heart and liver tissues were very low. LDH/CS (EC 4.1.3.7, citrate synthase) in skeletal muscle was the highest as 22.29. Specific activities of LDH in brain, eye and skeletal muscle were 56.45, 38.04 and 11.0 units/mg, respectively. The LDH isozymes in tissues were separated by polyacrylamide gel electrophoresis after immunoprecipitation with antiserum against $A_4,\;B_4$ eye-specific $C_4$ and liver-specific $C_4$. LDH $AC_4$ isozymes were detected predominantly in skeletal muscle, brain and eye tissues, and $B_4$ isozyme was detected in heart. Anodal eye-specific $C_4$ and cathodal liver-specific $C_4$ were coexpressed in A. hasta. The eye-specific $C_4$ isozyme showed higher activity in eye tissue, but liver-specific $C_4$ isozyme showed lower activity in liver. As a result, one part of molecular structures in $A_4\;and\;C_4,\;A_4\;and\;B_4$, and eye-specific $C_4$ and liver-specific $C_4$ were similar, but in $B_4\;and\;C_4$ were different with each other. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The LDH $A_4$ isozyme of skeletal muscle was purified in the fraction from elution with NAD+ containing buffer of affinity chromatography and eye-specific $C_4$ isozyme was eluted right after $A_4$, so the structure of eye-specific $C_4$ isozyme is similar to $A_4$. And LDH activity remained 35.22-43.47% as a result of the inhibition by pyruvate, the Michaelis-Menten constant values for pyruvate was 0.080-0.098 mM, and Vmax were 153.85 units, 35.09 units in skeletal muscle and eye, respectively. Also the $B_4$ isozyme was the thermo-stablest and $C_4$ was stabler than $A_4$ isozyme. The optimum pH of LDH was 6.5. The results mentioned above indicate that isozymes in tissues showed the properties between LDH $A_4\;and\;B_4$ isozyme as A. hasta was adapted to hypoxic conditions. Also LDH seems to function more effectively under anaerobic condition because LDH in skeletal muscle and eye tissues have high affinity for pyruvate.

• Journal of Life Science
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• v.19 no.2
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• pp.219-227
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• 2009

The antifatigue and endurance promoting properties of two Korean medicinal herb extracts and molasses with various mineral components were studied by evaluating forced-swimming capacity and biochemical parameters in ICR mice. The treatment groups were orally administered mineral beverages which were contained 6% sugar with the mixture of Maesil (Prunus mume fruit) extracts, Omija (Schisandra chinensis fruit) extracts and molasses for 4 weeks. The exercised forced-swimming tests were conducted after 28 days of beverage supplementation. The swimming times to exhaustion were longer 1.5${\sim}$2 times in group 6 and group 10 than control goup (Control: 93.2${\pm}$10.4 sec; Beverage 6; 190.8${\pm}$25.6 sec, Beverage 10; 173.6${\pm}$21.8 sec; p<0.05). Moreover, the activity of hexokinase (Control: 5.23${\pm}$0.38 ${\mu}mol$l/g tissue; Beverage 6: 5.99${\pm}$0.18 ${\mu}mol$/g tissue, Beverage 10: 6.13${\pm}$0.25 ${\mu}mol$/g tissue, p<0.05) and citrate synthase (control: 42.9${\pm}$1.87 ${\mu}mol$/g tissue; Beverage 6: 56.8${\pm}$3.98 ${\mu}mol$/g tissue, Beverage 10; 59.5${\pm}$3.09 ${\mu}mol$/g tissue, p<0.05) were also significantly higher than those of control group. Even if the treatment groups had long swimming than control group, there is no significant difference in the glycogen contents of gastrocnemus muscle or liver between the control group and each treatment group. This demonstrated an improvement in endurance. These results suggest that reported herbal beverage is very effective to combat fatigue, improve endurance and increase overall physical activity.

• Journal of Life Science
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• v.28 no.6
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• pp.656-662
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• 2018

Flavin-containing monooxygenases from Corynebacterium (cFMOs) were mutagenized based on homology modeling to develop variants with an enhanced indigoid production capability. The four mutants, F170Y, A210G, A210S, and T326S, which fused to a maltose-binding protein (MBP), were constructed, and their biochemical properties were characterized. Of these, purified MBP-T326S required a higher concentration of exogenous FAD (100 mM) than the wild-type MBP-cFMO for optimal activity and showed a 3.8-fold increase in the $k_{cat}/K_m$ value at $100{\mu}M$ FAD compared to that of MBP-cFMO at $2{\mu}M$ FAD. The indole oxygenase activities of MBP-T326S decreased to 63-77% compared to that of the MBP-cFMO In addition, MBP-T326S displayed a very low level of futile NADPH oxidase activities (21-24%) in the absence of a substrate. Mutant proteins except for T326S displayed similar $K_m$ and increased $k_{cat}/K_m$ values compared to the wild-type. MBP-F170Y and -A210S mutants showed elevated indole oxygenase activity higher than 3.1- and 2.9-fold, respectively, in comparison with MBP-cFMO. When indigoid production was carried out in LB broth with 2.5 g/l of tryptophan, Escherichia coli expressing cFMO produced 684 mg/l of indigo and 104 mg/l of indirubin, while cells harboring T326S produced 1,040 mg/l of indigo and 112 mg/l of indirubin. The results indicate that the production of indigo was 13% higher when compared to a previous report in which an E. coli expressing FMO from Methylophaga produced 920 mg/l of indigo. The protein engineering of cFMO based on homology modeling provided a more rational strategy for developing indigoid-producing strains.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.58-63
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• 2013

A taxonomic study was carried out to assess the phylogenetic characteristics of isolate JJM57 from marine red algae Laurencia sp. collected from intertidal zone in Jeju Island, South Korea. Comparative analysis of 16S rRNA gene sequence shows that this isolate belongs to the genus Oceanisphaera. It shows 98.02% and 97.7% sequence similarity with Oceanisphera litoralis DSM $15406^T$ and Oceanisphera donghaensis KCTC $12522^T$, respectively. Strain JJM57 is a Gram-negative, aerobic, moderately halophilic bacterium able to grow in different NaCl concentration ranges from 0.5 to 8.0% and at varying temperatures from 4 to $37^{\circ}C$. Sharing some of the physiological and biochemical properties with O. litoralis and O. donghaensis, JJM57 strain differs in the utilization of ethanol, proline, and alanine. The G+C contents of the strain JJM57 is 61.94 mol% and it is rich in $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2-OH, $C_{16:0}$, and $C_{18:1}$ ${\omega}7c$ fatty acids. The DNA-DNA relatedness data separates the strain JJM57 from other species such as O. litoralis and O. donghaensis. On the basis of these polyphasic evidences, present study proposed that strain JJM57 (=KCTC 22371 =AM983543 =CCUG 60764) represents a novel bacterial species of Oceanisphaera.

• Korean Journal of Organic Agriculture
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• v.26 no.2
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• pp.297-316
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• 2018

The golden apple snail (GAS, Pomacea canaliculata) is an invasive freshwater snail. The GAS was introduced in Korea without prior studies on the possibility of crop damage or its impact on the natural ecosystem. The freshwater apple snails can be found typically in ponds, rice paddies, irrigation canals, roadside ditches or slower portions of streams. In this study, we were carried out to investigate the assessment of physiological and ecological characteristics, environmental characteristics inhabited area in winter season and cause of massive death at one time of golden apple snails used for weed control in wet rice paddies. The GAS was introduced from Japan to Korea for commercial production as a dietary protein supplement. The golden apple snail was also used a recently for weed control in wet rice cultivation. The species of freshwater Pomacea snails is belonging to the genus Pomacea, family ampulariidae, order mesogastropoda, subclass pulmonata, class gastropoda, phylum mollusca. The GAS spread into irrigation ditches and natural waterways. It is now distributed in ponds and canals near rice fields of southern parts of the country and has overwintered. It increases its cold hardiness before winter. However, the physiological mechanism of cold hardiness in molluscs is poorly understood, especially in freshwater molluscs. Our results on physio-ecological characteristics of the Pomacea apple snail showed that the ratio of males to females was 1: 1.99~2.33. The daily growth was 87.7 mg in weight, 0.31 mm in height and 0.33 mm in width of the their shell. On the other hand, the golden apple snails were very high to resistance on drying condition and survived rate about 80% up to 3 months. The inhabitation of GAS was no statistical significant impacts on the water quality. An important property of aqueous solutions is agricultural water quality because it affects chemical and biochemical properties such as chemical reactions, equilibrium conditions, and biological toxicity. The death rate of weed control apple snails by Ostracoda (Stenocypris hislopi) was only 2.86% and 5.71% depending on the density. Therefore, GAS was not a direct death caused by Ostracoda (Stenocypris hislopi).

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.317-323
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• 2011

Strain A-3, an amylase-producing bacteria, was isolated from coastal seawater near Daecheon in the Republic of Korea. It was seen to possess a single polar flagella and grow well, on ASW-YP agar plates, at temperatures of between $20-37^{\circ}C$. However, it grew more slowly at the temperatures of $15^{\circ}C$ and $40^{\circ}C$. Similarly, it was observed to grow abundantly, in an Artificial Sea Water-Yeast extract-Peptone (ASW-YP) liquid medium, in a pH range of 6-9, but not grow at pHs of 4-5 and a pH of 10. Strain A-3 was noted as being close to Pseudoalteromonas phenolica O-$BC30^T$, Pseudoalteromonas luteoviolacea $NCIMB1893^T$, Pseudoalteromonas rubra $ATCC29570^T$, and Pseudoalteromonas byunsanensis $FR1199^T$, with 98.30%, 97.86%, 97.78%, and 97.25% similarities respectively, in its 16S rRNA sequence. A phylogenetic tree revealed that strain A-3 and P. phenolica O-$BC30^T$ belong to a clade. However, strain A-3 differed from P. phenolica O-$BC30^T$ in relation to a number of physiological characteristics. Strain A-3 exhibited no growth above 5% NaCl concentrations, no utilization of D-glucose, D-mannose, D-maltose, or D-melibose, and no lipase (C-14) activity. All of these properties strongly indicate that strain A-3 is distant from P. phenolica O-$BC30^T$ and thus led us to name it Pseudoalteromonas sp. A-3. Pseudoalteromonas sp. A-3 produces ${\alpha}$-amylase throughout growth. Maximal amylase activities of 144.48 U/mL and 149.20 U/mL were seen at pH 7.0 and $37^{\circ}C$, respectively. Pseudoalteromonas sp. A-3's high, stable production of ${\alpha}$-amylase in addition to its biochemical features, such as alkalitolerance, suggest that it is a good candidate for industrial applications.