• Proceedings of the Korean Society of Toxicology Conference
• /
• 1997.05a
• /
• pp.65-74
• /
• 1997

By using guinea pig lung mast cells, this study aimed to examine the effects of Aloe component(NY945) on the mediator releases caused by mast cell activation, and also aimed to assess the effects of NY945 on the mechanism of mediator releases in the mast cell activation. We partially purified mast cells from guinea pig lung tissues by using the enzyme digestion, the rough and the discontinuous density percoll gradient method. Mast cells were sensitized with $IgG_1$ (anti-OA) and challenged with ovalbumin. Histamine was assayed by fluorometric analyzer, leukotrienes by radioimmunoassay The phospholipase D activity was assessed more directly by the production of labeled phosphatidylethanol or phosphatidylbutanol which was produced by phospholipase D-mediated transphosphatidylation in the presence of ethanol or butanol. The amount of mass 1,2-diacylglycerol was measured by the [$^3H$]1,2-diacylgycerol produced when prelabeled with [$^3H$]myristic acid. In the mast cells prelabeled with L-[$^3H$]methyl methionine the phospholipid methylation was assessed by measuring the incorporation of the [$^3H$]methyl moiety into phospholipids. Pretreatment of NY945(10$\mu$g) significantly decreased histamine and leukotrienes releases during mast cell activation. The decrease of histamine release was stronger than that of leukotrienes during mast cell activation. The phospholipase D activity increased by the mast cell activation was decreased by the dose-dependent manner in the pretreatment of NY945. The amount of mass 1,2-diacylglycerol produced by activation of mast cells were decreased in the pretreatment of NY945. NY945 pretreatment strongly inhibited the incorporation of the [$^3H$]methyl moiety into phospholipids. The data suggest that NY945 purified from Aloe inhibits in part an increase of 1,2-diacylglycerol which is produced by activating mast cells with antigen-antibody complexes which is mediated via phosphatidylcholine-phospholipise D and phosphatidylinositole-phospholipise C systems, and then followed by the inhibition of histamine release. Furthermore, NY945 reduces the phosphatidylcholine production by inhibiting the methyltransfsrase I and II, which decrease the conversion of phosphatidylcholine into arachidonic acid and inhibits the production of leukotrines.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.4 no.2
• /
• pp.249-255
• /
• 2001

Methylmalonic acidemia is a rare congenital autosomal recessive metabolic disease. It is caused by blocking in the pathways of isoleucine, valine, threonine, methionine, cholesterol and odd-chain fatty acids to succinyl CoA, resulting in the increase of L-methylmalonyl CoA and methylmalonic acid. In most cases, there are symptoms such as recurrent vomitings, lethargy and laboratory abnormalities including metabolic acidosis and hyperammonemia from the neonatal period. We had a 6-month-old infant with methylmalonyl acidemia who presented with recurrent vomiting episodes since 3 months of age, failure to thrive and developmental delay. The laboratory findings showed hyperammoninemia and ketotic metabolic acidosis. Plasma amino acid analysis showed nonspecific finding. Urine organic acid ananysis by gas chromatography and mass spectrometry detected large amount of methylmalonic acid excreted in the urine. We restrained the supply of protein in the amount of 1~1.5 g/kg of body weight a day using leucine, isoleucine and valine-r-estrained milk and administered vitamine $B_{12}$, in the amount of 1mg per day. During the follow-up in the outpatient clinic, He could control his head and showed increased muscle strength.

• Journal of Fisheries and Marine Sciences Education
• /
• v.29 no.3
• /
• pp.888-898
• /
• 2017

Tenebrio molitor larvae, also known as yellow mealworms (MW), are rich in protein and lipid and can serve as a potential alternative protein and energy source in commercial aquafeeds. Therefore, this study attempts to evaluate the effects of different drying methods on the nutritional value of MW meal. For this, live MW were cold-anaesthetized before being subjected to three different types of drying methods, including freeze-drying, oven-drying at $60^{\circ}C$ and air-drying at room temperature for three days, and compared for proximate composition and energy content. An in-vivo digestibility test was then conducted to evaluate the nutrient digestibility of MW meal in diets for rockfish, Sebastes schlegeli. A test diet was prepared by mixing the MW meal with a reference diet (Ref) in a 30:70 ratio with chromium oxide as an inert marker at the inclusion level of 0.5%. Rockfish with mean body weight of 150 g were stocked into a fecal collection system equipped with fiberglass tanks of 400 L capacity. Each group of fish was fed one of the experimental diets to apparent satiation for 4 weeks. The results of the proximate analysis showed that drying methods had no significant effect on crude protein, crude lipid, ash and energy contents of MW. Despite being a rich source of protein and lipid, MW meal was deficient in certain amino acids, particularly methionine, and highly unsaturated fatty acids, particularly 22:6n-3 (DHA) and 20:5n-3 (EPA). MW meal showed high digestibility values for protein (93%), lipid (97%) and energy (88%). These results may indicate that MW meal is a nutritious and acceptable feed ingredient, with comparable digestibility values to conventional animal and plant feedstuffs such as fish meal and soybean meal, in practical diet for rockfish at grower stage.

• BMB Reports
• /
• v.32 no.3
• /
• pp.299-305
• /
• 1999

Protein carboxyl O-methyltransferase (PCMT) catalyzes the transfer of a methyl group from Sadenosyl-L-methionine to free carboxyl groups of methyl-accepting substrate proteins. Two isozymes were separated by DEAE-Sephacel chromatography from porcine brain cytosol and designated PCMT I and II. Isozymes I and II were further purified by adenosyl homocysteine-Sepharose 4B and Superose HR 12 chromatography. The molecular weights of the purified PCMT I and II were determined by mass spectrometry to be 20,138 Da and 25,574 Da, respectively. The two enzymes displayed different isoelectric points; 7.9 for PCMT I and 5.3 for PCMT II. Isozymes I and II exhibited similar substrate specificities when tested with various methyl-accepting proteins. Myelin basic protein, a component of myelinated neurons, was found to be an excellent methyl-accepting substrate for both PCMT isozymes with different $K_m$ values, $21.1\;{\mu}M$ for PCMT I and $10.6\;{\mu}M$ for PCMT II. The PCMT activity and methyl-accepting capacity displayed similar distribution in the various brain regions with an exception of the lower values in the cerebellum. The overall distribution may relate to a general function of protein repair by PCMT in the brain.

• Journal of Animal Science and Technology
• /
• v.58 no.10
• /
• pp.36.1-36.5
• /
• 2016

Background: Liquid feeding system has been introduced to domestic swine farms, but negative cognition about liquid feeding system has been remained for feed waste decay related with poor management and microbial contamination. For these reasons, this study was conducted to evaluate the effects of feeding method in lactating sows. Methods: A total of 30 mixed-parity (average 4.13) lactating sows (Yorkshire ${\times}$ Landrace) with an initial BW of $218.8{\pm}19.5kg$ was used in a 3 week trial. Sows were allotted to 1 of 2 treatments in a completely randomized design by their body weight, backfat thickness, parity and alive litter weight. One of treatments was dry feeding and the other was liquid feeding (water to feed ratio, 1:1). Experimental diets contained 3265 kcal ME/kg, 12.6 % CP, 5.76 % EE, 1.09 % total lysine, 0.25 % total methionine, as fed basis. Results: Dry feeding treatment had high body weight loss rather than liquid feeding treatment (P = 0.04). Dry feeding treatment had tendency to increase litter weight at 21d of lactation (P = 0.06) and litter weight gain (P = 0.04) during lactation period (0-3 week). Sows fed dry feeding method made milk containing high content of casein and total solid rather than sows fed liquid feeding method (P = 0.04). In addition, dry feeding treatment had tendency to higher content of milk fat, protein and solid not fat on 21d of lactation (P = 0.07). Sows fed dry feeding type also showed higher milk energy content in milk of 21d lactation (P = 0.05). Furthermore, liquid feeding treatment showed high occurrence in feed waste during lactation period (P <0.01). Conclusion: Dry feeding method was more suitable feeding method to lactating sows under high temperature environment like lactating barn.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.7
• /
• pp.1100-1107
• /
• 2007

The effects of offering broilers phosphorus-adequate diets containing 10.0 and 11.8 g/kg lysine, without and with 500 FTU/kg exogenous phytase, on growth performance and nutrient utilisation were determined. Each of the four experimental diets was offered to 6 replicates of 10 birds from 7 to 28 days of age. Effects of treatment on performance, apparent metabolisable energy, apparent ileal digestibility of amino acids and bone mineralisation were examined. Both additional lysine and phytase supplementation improved (p<0.05) weight gain and feed efficiency, with interactions (p<0.05), as phytase responses were more pronounced in lysine-deficient diets. Phytase improved (p<0.05) apparent metabolisable energy, which was independent of the dietary lysine status. Bone mineralisation, as determined by percentage toe ash, was not affected by treatment, which confirms the phosphorus-adequate status of the diets. Phytase increased (p<0.05) the apparent ileal digestibility of the sixteen amino acids assessed. Unexpectedly, however, the dietary addition of 1.8 g/kg lysine, as lysine monohydrochloride, increased (p<0.05) the ileal digestibility of lysine per se and also that of isoleucine, methionine, phenylalanine, valine, aspartic acid, glutamic acid and tyrosine. In addition, there were significant interactions (p<0.05) between additional lysine and phytase supplementation for arginine, lysine, phenylalanine, aspartic acid, glutamic acid, glycine and serine digestibilities, with the effects of phytase being more pronounced in lysine-deficient diets. The possible mechanisms underlying the increases in amino acid digestibility in response to additional lysine and the interactions between lysine and microbial phytase in this regard are discussed. Also, consideration is given to the way in which phytate and phytase may influence ileal digestibility of amino acids.

• Fisheries and Aquatic Sciences
• /
• v.14 no.1
• /
• pp.1-10
• /
• 2011

This study was conducted to obtain hydrolysates with potent antioxidative activity from rockfish skin gelatin. Gelatin was extracted under high temperature/high pressure using a two-step enzymatic hydrolysis with commercial enzymes such as Alcalase, Flavourzyme, Neutrase, and Protamex. The second rockfish-skin gelatin hydrolysate (SRSGH) was prepared by further incubating the first gelatin hydrolysate (FRSGH), which had been hydrolyzed with Alcalase for 1-h (FRSGH-A1), with Flavourzyme for 2-h (SRSGH-F2). The second gelatin hydrolysate showed higher antioxidative activity of 3.72 as measured by a Metrohm Rancimat and superior angiotensin I-converting enzyme (ACE) inhibiting activity of 0.82 mg/mL. Compared with the gelatin, the relative proportion in SRSGH-F2 was markedly decreased in the 100-kDa peak, whereas it was increased in that less than 100-kDa. The amino acid composition of SRSGH-F2 was rich in glycine (25.9%), proline (10.8%), alanine (9.1%), and glutamic acid (9.1%). In contrast, it was poor in cystine (not detected), methionine (1.6%), tyrosine (0.4%), hydroxylysine (0.9%), and histidine (0.9%). In recent years, demand for natural functional foods has been increasing, and SRSGH-F2 can be used as a functional food ingredient in the food industries. However, further detailed studies on SRSGH-F2 with regard to its antioxidant activity in vivo and the various antioxidant mechanisms are needed.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.53 no.3
• /
• pp.464-470
• /
• 2020

This study was conducted to replace fish meal (FM) with three plant proteins (soybean meal, soy protein concentrate, and wheat gluten) in diets for growing olive flounder Paralichthys olivaceus. The control diet was formulated to contain 65% sardine FM and four other replacement diets were formulated to replace FM with the plant proteins by 25, 30, 35 and 40% (designated FM25, FM30, FM35 and FM40, respectively). The replacement diets were added with three essential amino acids (lysine, methionine and threonine) to meet their requirements for the fish. Olive flounder (initial average weight, 96.8±0.2 g) were randomly distributed into 20 tanks (425 L each) at a density of 25 fish per tank. Four replicate groups of fish were fed one of the diets two times daily for 15 weeks. At the end of the feeding trial, no significant differences were found among all the fish groups in growth performance, feed utilization, nonspecific immune responses and hematological health parameters. Thus, this result indicates that the plant proteins with the three limiting amino acids could replace FM up to 40% in diets for growing olive flounder.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.9
• /
• pp.1359-1366
• /
• 2010

A search of the Streptomyces avermitilis genome reveals that its closest homologs are several O-methyltransferases. Among them, one gene (viz., saomt5) was cloned into the pET-15b expression vector by polymerase chain reaction using sequence-specific oligonucleotide primers. Biochemical characterization with the recombinant protein showed that SaOMT5 was S-adenosyl-L-methionine-dependent Omethyltransferase. Several compounds were tested as substrates of SaOMT5. As a result, SaOMT5 catalyzed O-methylation of flavonoids such as 6,7-dihydroxyflavone, 2',3'-dihydroxyflavone, 3',4'-dihydroxyflavone, quercetin, and 7,8-dihydroxyflavone, and phenolic compounds such as caffeic acid and caffeoyl Co-A. These reaction products were analyzed by TLC, HPLC, LC/MS, and NMR spectroscopy. In addition, SaOMT5 could convert phenolic compounds containing ortho-dihydroxy groups into O-methylated compounds, and 6,7-dihydroxyflavone was known to be the best substrate. SaOMT5 converted 6,7-dihydroxyflavone into 6-hydroxy-7-methoxyflavone and 7-hydroxy-6-methoxyflavone, and caffeic acid into ferulic acid and isoferulic acid, respectively. Moreover, SaOMT5 turned out to be a $Mg^{2+}$-dependent OMT, and the effect of $Mg^{2+}$ ion on its activity was five times greater than those of $Ca^{2+}$, $Fe^{2+}$, and $Cu^{2+}$ ions, EDTA, and metal-free medium.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.224-224
• /
• 2017

In plants, cysteine(Cys) is decisive for protein and glutathione that acts as an indispensable sulfur grantor for methionine and many other sulfur containing secondary products. Cys formation is involved in the consecutive two reactions using two enzymes-serine acetyl transferase (SAT) and O-acetylserine (thiol)lyase (OASTL) and appeared in plant cytosol, chloroplast and mitochondria. OASTL is able to produce mimosine with 3-hydroxy-4-pyridone (3H4P) in lieu of $H_2S$ for Cys. In this report, we describe the first time cloning, purification and characterization of cytosolic(cy)OASTL from M. pudica and its expression in Escherichia coli and try to find out the cross link between this OASTL and the mimosine formation and to elucidate the metabolic role of cy-OASTL in M. pudica. The purified recombinant protein was 34.7 KDa. The optimum reaction pH and temperature was 6.5 and $50^{\circ}C$, respectively. The Michaelis constant (Km) and the Vmax value of the enzyme was $252{\pm}25{\mu}M$ and $57{\pm}3{\mu}M\;cysteine\;min^{-1}\;{\mu}g\;protein^{-1}$ for sulfide and $159{\pm}21{\mu}M$ and $58{\pm}2.4{\mu}M\;cysteine\;min^{-1}\;{\mu}g\;protein^{-1}$ for OAS subsequently. After cleaving the His-tag, we tried to observe cy-OASTL to form mimosine with appropriate substrate but it was not successful. It may be concluded that cy-OASTL of the present study is only Cys specific, not mimosine.