• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2002.10a
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• pp.18-25
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• 2002

The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

• Current Research on Agriculture and Life Sciences
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• v.7
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• pp.99-107
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• 1989

The study was conducted to obtain some basic information to establish the system of performance-tests and selection of honeybee queens(Apis mellifera) under Korean circumstances, Colony performances were tested with thirty colonies of Apis mellifera at two apiaries in Taegu, Korea from September, 1988 to August 1989. The results of performance-testing on the colonies are summarized as follows : The colony weight measured before wintering was averaged $23.6{\pm}1.90kg$ and the colony weight was decreased by $2.9{\pm}0.82kg$ in average during winter season. Thirteen colonies were entered in two story hive from thirty single box colonies from April 17 to May 5, 1989 with increase of bee population and, consequently, the ability of enter-supers of the colonies apperared to be low. The ability of collecting pollen was measured to be $14.8{\pm}2.15gr$ per colony during 24 hours in April, and the number of swarm cells was counted $12.5{\pm}3.43$ cells per colony in aveage. Tendency to use propolis appeared to be moderate, and the number of returning foragers for a minute per colony was counted $108.7{\pm}18.31$ bees in average. Brood area was measured $2,464{\pm}628,67cm^2$ per colony in the post nectar flow season of acasia, and 30.8 percent of the colonies appeared to be infected with chalkbrood disease, The amount of honey production was $14.9{\pm}8.49kg$ per colony, which was harvested two times during the main nectar flow season of acasia.

• The Korean Journal of Nuclear Medicine
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• v.27 no.2
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• pp.223-232
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• 1993

Technetium labeled isonitrile analogues are widely used as myocardial perfusion imaging agents. We synthesized and characterized a new isonitrile compound, ethyl 3-isocyanobutyrate(EIB). Proton and $^{13}C$ NMR spectroscopy and thin layer chromatography with a $C_{18}$ coat was performed. EIB was easily labeled with $^{99m}TcO_4^-$- with sodium dithionite. The labeling efficiency measured by RP-HPLC was over 95%. The labeled product was stable with dilution in normal saline and with prolonged incubation at room temperature. There was no formation of secondary products or free $^{99m}TcO_4^-$. In vivo kinetics study of $^{99m}Tc$ (I) labeled EIB in rabbits showed adequate myocardial uptake, good contrast against lung background, and relatively rapid liver clearance. The heart to lung ratio was over 2.5 and the heart to liver ratio was approximately from 0.4 to 5 at 60 minutes post injection. Hepatic clearance of $^{99m}Tc-MIBI$ was faster ($t_{1/2}$=6 minutes) than that of $^{99m}Tc-MIBI$. In vivo kinetics observed in dog was similar to that in rabbit but there was faster gallbladder filling, and thus lower liver background. SPECT imaging of the canine myocardium showed favorable imaging characteristics. However, biodistribution in mice demonstrated a myocardial % injected dose/organ of less than 0.1%. This was thought to be due to interspecies difference in plasma esterase activity. In human plasma, $^{99m}Tc$ ( I ) labeled EIB was stable for at least 2 hours, without production of secondary products by HPLC. We conclude that ethyl 3-isocyanobutyrate may be a potential new myocardial perfusion imaging agent and deserves further investigation as to its usefulness for clinical use.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.6
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• pp.1216-1221
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• 2012

This study is aimed to investigate microbiological contamination of Angelica gigas Nakai. A total of 111 samples including root, soil, and irrigation water were collected from farms and market to detect aerobic bacteria, Bacillus cereus, coliform, Escherichia coli, Listeria monocytogenes,. Salmonella spp., and Staphylococcus aureus. The contaminations of aerobic bacteria, coliform, and Bacillus cereus in the root during cultivation were found 6.71 log CFU $g^{-1}$, 4.13 log CFU $g^{-1}$, and 3.54 log CFU $g^{-1}$, respectively. The contamination of coliform and B. cereus were detected in all steps from harvesting to processing, with the highest count recorded from the cutting step. In marketing, the contaminations of aerobic bacterial, coliform, and B. cereus were 5.5~6.0 log CFU $g^{-1}$, 2.4~2.6 log CFU $g^{-1}$, and 3.5~4.0 log CFU $g^{-1}$, respectively. Listeria monocytogenes, Salmonella spp, and Staphylococcus aureus were not detected in any of samples. This result indicated that hygienic soil management and post harvest management should be performed to reduce the contamination of hazard microorganisms and to produce safe agro-products.

• Applied Biological Chemistry
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• v.37 no.6
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• pp.433-440
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• 1994

The exposure of Clamydomonas reinhardtii to low temperatures resulted in an accumulation of cellular pyruvate that dissipated when the chilled cells returned to ambient temperature. The dissipation of pyruvate accumulation was accompanied by an increase in the production level of superoxide radicals $(O_2^-)$ in cells. The formation of $O_2^-$ at an excessive level during the post-chilling period was apparently countered by a substantial activation of superoxide dismutase (SOD). All these results are similar to those observed previously in rice seedlings subjected to the cold-treatment, implicating that a common mechanism is probably underlying for the primary processes of chilling injury both in higher plants and in algae. It was also observed that the activation of Mn-containing SOD contributes the major share in the increase of SOD activity of whole algal cells. Because Mn-SOD is present only in mitochondria, the observation corroborates the concept that the $O_2^-$ scavenging enzyme would be induced to cope with the cold treatment-caused adverse situation in mitochondria where the toxic active oxygen is produced at rates far exceeding the normal rate.

• Journal of Biosystems Engineering
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• v.6 no.2
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• pp.33-47
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• 1982

To modernize the conventional rice post production technology and reduce grain losses, a transition toward the wet-paddy threshing system has been strongly demanded. The head-feeding type thresher with pneumatic separation has been used dominantly for threshing dried-paddy, but some adverse effects in separation performance for threshing wet-paddy is encounterred. In order to solve the problems, the development of thresher with an additional oscillating sieve to the conventional pneumatic separation has been recommanded. This study was intended to evaluate the separating performance of thresher with oscillating sieve which was attached additionally to the conventional auto-thresher equipped with separation system of blower and suction fan. For different feed rates and rice varieties, wet-and dry-material were tested with threshers attached with and without oscillating sieve. Results of the study are summarized as follows: 1. When the feed rates were 480 and 640 kg/hr, there was no statistically significant difference in power reqirements between the threshers with and without an additional sieve device for both dry-and wet-threshing. However, when the feed rate was 960 kg/hr, power requirements of thresher without sieve were greater for wet-paddy threshing than the thresher with the additional sieve separator by about 20% points. 2. With additional oscillating sieve device, the ratios of total weights of whole grains including grains with branch let and damaged grains to the total output did not show statistical difference among the feed rates. However, with pneumatic separation the ratio was decreased as the level of feed rate increased. 3. The total amount of grains with branchlet (including broken panicle) increased with the moisture content. For both the wet-and dry-material threshing with the additional oscillating sieve, the percent of grains with branchlet to the total output decreased greatly as the feed rate increased. 4. The output of the damaged grains increased as moisture content decreased. Especially, for the dry-paddy threshing, the additional sieve separating device produced more damaged grains than the pneumatic separation at all feed rates. 5. Generally, for dry paddy threshing, the separating performance of the thresher with the additional sieve device was better at all feed rates, showing greater difference with increasing feed rates. 6. Separating losses were greater with the pneumatic than sieve separation for both the wet-and dry-threshing. 7. The overall comparison of separating performance of threshers tested with and without an additional sieve device showed that the former was more effective than the latter for the dry-material threshing. However, for the wet-paddy threshing, the separation performance with a sieve device was better than the pneumatic only when the feed rate was high.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.155-162
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• 2005

The present study evaluated whether an exogenous antioxidants, taurine and $\alpha$-tocopherol, could, when added to the freezing extender, improve the post-thaw sperm characteristics, function, the level of reactive oxygen species (ROS) generation, and the level of lipid peroxidation (LPO) in frozen-thawed porcine semen. CASA (computer-aided sperm analysis), HOST (hypoos-motic swelling test), chemiluminescence using luminol and lucigenin and the detection of malondialdehyde for LPO was performed in frozen-thawed porcine sper-matozoa. The results obtained in these studies are as follows. While no beneficial effects of taurine and $\alpha$-tocopherol supplementation were visible in motility, viability, acrosome reaction, tail swelling patterns, and the generation of $O^{2-}$ of frozen-thawed porcine sper-matozoa, $H_{2}O_{2}$ was decreased by all treatments except taurine 50mM treatment. In conclusion the taurine and $\alpha$-tocopherol treatments during freezing reduced generation of reactive oxygen species and production of malondialdehyde in frozen-thawed porcine semen, and the ROS savangers may minimize various damages of spermatozoa during freezing.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.3
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• pp.346-351
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• 2002

The effect of fetal number (single or twin) on plasma concentrations of progesterone, estradiol $17{\beta}$, cortisol, prolactin, growth hormone, triiodothyronine, thyroxine and insulin around parturition (periparturient period) were studied on ten $Alpine{\times}Beetle$ crossbred goats in their first to third lactation. The hormone profiles were studied on days -20, -15, -10, -5, -4, -3, -2, -1 prior to kidding and on day 0 and +1, +2, +3, +4, +5, +10, +15, +20 days postkidding. Plasma progesterone levels were significantly (p<0.01) higher in twin bearing goats comparison to single bearing goats during all the days of sampling. The decline in progesterone concentration from day 20 to day 1 before kidding was 56% in twin and 42% in single bearing goats. In single bearing goats plasma estradiol $17{\beta}$ was significantly (p<0.01) higher during prekidding days compared to twin bearing goats. The level of estradiol $17{\beta}$ was highest on the day of kidding in both the groups. The plasma prolactin level in twin bearing goats from day 10 to day 1 prepartum was higher as compared to single fetus bearing goats. However there was abrupt increase in prolactin level on the day of kidding in both the groups. The plasma growth hormone levels were significantly (p<0.01) higher in twin compared to single bearing goats. On the day of kidding growth hormone levels were significantly (p<0.01) higher in twin as compared to single bearing goats (1.40 vs. 0.95 ng/ml). In twin bearing goats plasma cortisol values from day 5 till the day of kidding remained elevated and the levels on the day of kidding was significantly highest in both the groups. The levels of triiodothyronine ($T_3$) were significantly higher (p<0.01) during all the periods of sampling in single compared to twin bearing goats. Plasma thyroxine ($T_4$) was significantly (p<0.01) lower in twin compared to single bearing goats. In single bearing goats plasma insulin levels were significantly (p<0.01) higher than twin bearing goats during prepartum period however during post partum period the levels in both the groups remained similar. It can be concluded that number of fetuses is having significant influence on the hormone profile during periparturient period.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.3
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• pp.258-264
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• 2020

This study investigated the effect of variation in the number of somatic-cell-cloned embryos and their developmental stage at transfer on pregnancy, as well as the influence of the estrus status of recipient pigs on in vivo development of cloned porcine embryos after embryo transfer. For somatic cell nuclear transfer (SCNT), fibroblast cells were obtained from a male porcine fetus. Recipient oocytes were collected from prepubertal gilts at a local abattoir and then cultured. After SCNT, reconstructed embryos of different numbers and developmental stages were transferred into recipient pigs. The developmental stage of the cloned embryos and the number of transferred embryos per surrogate showed no significant differences in terms of the resulting cloning efficiency. However, the pregnancy rate improved gradually as the number of transferred cloned embryos was increased from 100-150 or 151-200 to 201-300 per recipient. In pre-, peri-, and post-ovulation stages, pregnancy rates of 28.6%, 41.8%, and 67.6% and 16, 52, and 74 offspring were recorded, respectively. The number of cloned embryos and estrus status of the recipient pig at the time of transfer of the cloned embryo affect the efficiency of pig production; therefore, these variables should be particularly considered in order to increase the efficiency of somatic cell pig cloning.

• Reproductive and Developmental Biology
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• v.29 no.2
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• pp.83-91
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• 2005

Somatic cell nuclear transfer in cattle has limited efficiency in terms of production of live offspring due to high incidence of fetal failure after embryo transfer to recipients. Such low efficiency of cloning could possibly arise from abnormal and poorly developed placenta. In the present study the placental proteome in late pregnancy established from in vitro fertilization (IVF) and nuclear transfer (NT) was analysed. Proteome alternation was tested using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI- TOF). Comparing placenta from NT embryos to those from IVF counterparts, significant changes in expression level were found in 18 proteins. Of these proteins 12 were not expressed in NT placenta but expressed in IVF counterpart, whereas the expression of the other 6 proteins was limited only in NT placenta. Among these proteins, cytokeratin 8 and vimentin are considered to be involved in regulation of post-implantation development. In particular, cytokeratin 8 and vimentin may be used as makers for placental development during pregnancy because their expression levels changed considerably in NT placental tissue compared with its IVF counterpart. Data from 2-DE suggest that protein expression was disorientated in late pregnancy from NT, but this distortion was eliminated with progression of pregnancy. These findings demonstrate abnormal placental development during late pregnancy from NT and suggest that alterations of specific placental protein expression may be involved in abnormal function of placenta.