L-carnitine promotes mitochondrial ${\beta}$-oxidation of long chain fatty acids and their subsequent transport across the inner mitochondrial membrane. Although the role of L-carnitine in fatty acid metabolism has been extensively studied, its role in live performance and carcass responses of commercial broilers is less understood. The objective of this research was to determine if Lcarnitine fed at various levels in diets differing in CP and amino acids impacted on live performance and carcass characteristics of commercial broilers. Two floor pen experiments were conducted to assess the effect of dietary L-carnitine in grower diets. In Exp. 1, Ross${\times}$Hubbard Ultra Yield broilers were placed in 48 floor pens (12 birds/pen) and fed common diets to d 14. A two (0 or 50 ppm Lcarnitine) by three (173, 187, and 202 g/kg CP) factorial arrangement of treatments was employed from 15 to 35 d of age (8 replications/treatment). An interaction (p<0.05) in carcass yield indicated that increasing CP (187 g/kg) resulted in improved yield in the presence of L-carnitine. Increasing CP from 173 to 202 g/kg increased (p<0.05) BW gain and decreased (p<0.05) feed conversion and percentage abdominal fat. Feeding dietary L-carnitine increased back-half carcass yield which was attributable to an increase (p<0.05) in thigh, but not drumstick, yield relative to carcass. In Exp. 2, $Ross{\times}Ross$ 708 broilers were fed common diets until 29 d. From 30 to 42 d of age, birds were fed one of seven diets: i) 200 g/kg CP, 0 ppm L-carnitine; ii) 200 g/kg CP, 40 ppm L-carnitine; iii) 180 g/kg CP, 0 ppm L-carnitine; iv) 180 g/kg CP, 10 ppm L-carnitine; v) 180 g/kg CP, 20 ppm L-carnitine; vi) 180 g/kg CP, 30 ppm L-carnitine; and vii) 180 g/kg CP, 40 ppm L-carnitine (6 replications of 12 birds each). BW gain, feed conversion, mortality (30 to 42 d), and carcass traits (42 d) were measured on all birds by pen. There were no treatment differences (p<0.05). However, the addition of 40 ppm L-carnitine in the 200 g CP/kg diet increased (p = 0.06) thigh yields relative to BW in comparison to birds fed diets without L-carnitine, which was further confirmed via a contrast analysis (0 vs. 40 ppm L-carnitine in the 200 and 180 g CP/kg diets; p<0.05). These results indicated that dietary L-carnitine may heighten metabolism in dark meat of commercial broilers resulting in increased relative thigh tissue accretion without compromising breast accretion.
Acidic chitinases from the gizzards of a broiler were purified to homogeneity, using precipitation with $(NH_{4})_{2}SO_{4}$, ion exchanger chromatography, gel filtration, chromatofocusing and hydrophobic interaction chromatography. The enzymes, GAC1 and GAC2, were purified 180- and 194- folds with a recovery of 4.9% and 2.7%, respectively. The molecular mass of GAC1 and GAC2 were 48.2 kDa and 57.8 kDa, respectively. Chromatofocusing resulted in a pI of 3.1 for both enzymes. The purified enzymes were endochitinases that were devoid of ${\beta}-N-acetylglucosaminidase$ and lysozyme activity. Kinetic studies using $[^3H]chitin$ indicate that GAC1 has a $K_m$ and $V_{max}$ of 1.97 mg/ml and 185 mg/mg protein/h, respectively. The GAC2 has a $K_m$ and $V_{max}$ of 0.42 mg/ml and 92.3 mg/mg protein/h, respectively at optimal pH and temperature (pH 5.0 and $60^{\circ}C$). When the pentamer and hexamer of N-acetylglucosamine (GlcNAc) were used as a substrate, the major product by GAC1 was the dimer of GlcNAc with a differential accumulation of the monomer and trimer, depending upon the substrate. However, the GAC2 produced the dimer and trimer in an equal quantity, regardless of the substrate used. The first 9 $NH_2-terminal$ amino acid residues of the purified gizzard chitinase GAC1 and GAC2 shared a 100% homology. The first 25 $NH_2-terminal$ amino acid residues of GAC1 also shared 55-60% homology with animal chitinases and some animal proteins, such as whey protein and oviduct-specific proteins. However, little homology was found with either microbial and plant chitinases, or egg white lysozyme.
Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl ${\beta}$-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.
Kim, Tae-Hoon;Ahn, Hee-Young;Kim, Young-Wan;Sim, So-Yeon;Seo, Kwon-Il;Cho, Young-Su
Journal of Life Science
/
v.27
no.12
/
pp.1470-1478
/
2017
The aim of this study was to investigate biological activity and biochemical properties of extracts from Bacillus subtilis-fermented silkworm (Bombyx mori L., SP) powder of different origin (Buan, Namwon, and Boeun). An additional aim was to determine the inhibition of cancer cell (B16-F10, HT-29, LNcaP, and MCF-7) proliferation and nitric oxide (NO) production from lipopolysaccharide (LPS)-induced RAW264.7 cells. Biological activities (${\alpha},{\alpha}^{\prime}$-diphenyl-${\beta}$-picrylhydrazyl [DPPH], free radical scavenging activity, fibrinolytic activity, antiproliferation activity, and anti-inflammatory activity) and biochemical properties (compositional amino acid contents, and mineral contents) were examined in water extracts from silkworm powder and B. subtilis-fermented silkworm powder. The highest amino acid contents were detected in Buan silkworm powder (BU). After fermented, the highest contents were found in B. subtilis-fermented Buan silkworm powder (BBO). The major minerals detected were K, Ca, and Mg. Rates of these minerals, especially those of Na increased after fermented. DPPH radical scavenging activity and fibrinolytic activity were stronger in the fermented group than non-fermented group. DPPH radical scavenging activity and fibrinolytic activity were highest in the extract from BBO. The inhibition activities of LNcaP and MCF-7 cells viability were significantly decreased in the BBO, and there was no inhibition activity in other cancer cells (B16-F10 and HT-29). An SRB assay of the cell viability of RAW 264.7 cells exposed to extracts of silkworm powder and B. subtilis-fermented silkworm powder revealed no toxicity in any of the groups. Compared with the LPS-treated group, the biggest reduction in NO production was detected in the BBO group. Based on these results, extracts from Boeun silkworm powder fermented with B. subtilis could be a candidate material as a dietary supplement for use in healthy functional foods.
Journal of the Korean Society of Food Science and Nutrition
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v.42
no.5
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pp.721-729
/
2013
To improve the quality and functionality of the low salt squid Jeot-gal, extracts from three types of medicinal and edible plants (bay leaf, green tea, pine needle) were added. The quality characteristics, bioactivities, and shelf-lives of these preparations were determined at three different fermentation temperatures. The pH decreased more rapidly at higher temperatures, while the amount of volatile basic nitrogen (VBN), total viable cells, and amino nitrogen ($NH_2$-N) increased. The shelf-lives of Jeot-gal with natural plant extracts at $10^{\circ}C$ were 34~35 days, similar to the control. The major free and compositional amino acids of Jeot-gal were glutamic acid, proline, and alanine, while the major nucleotides (and related compounds) were hypoxanthine and inosine. In bioactivity assays, samples supplemented with plant extracts showed higher bioactivities than the control. The DPPH radical scavenging activity of ethanol extracts from Jeot-gal were stronger than the water extracts; in contrast, the water extracts were stronger for hydrogen peroxide scavenging activity. However, superoxide dismutase (SOD)-like activity and ${\beta}$-glucuronidase inhibitory activity were moderately low at 20 mg/mL. Based on sensory evaluation results, the quality of low salt squid Jeot-gal with natural plant extracts is similar to the control. Therefore, low salt squid Jeot-gal with natural plant extracts can be commercialized as a functional fermented food.
Although barley $\alpha$-amylase isozyme 1 (AMY1) and 2 (AMY2) share up to 80% of amino acid sequence identity, their enzymatic properties differ remarkably. In this study, the 42nd alanine residue of AMY2 was replaced with another random amino acid via saturation mutagenesis. Eight out of 370 recombinant E. coli cells showing enhanced starch-hydrolyzing activity were characterized as possessing the same proline residue instead of alanine. Even though the specific activity of AMY2-A42P is reduced to 81% of wild-type, its expression level and purification yield were enhanced by approximately 2 and 4 times that of AMY2, respectively. Characterization of its enzymatic properties confirmed that AMY2-A42P is similar to that of wild-type. However, its specificity to starch substrates is likely to be intermediate between AMY1 and AMY2.
Purpose : We hypothesized that the persisting bronchial hyperresponsiveness (BHR) of adolescents with asthma remission may be controlled mainly by genetic factors, and the BHR of symptomatic asthma by airway inflammation. ${\beta}_2$-adrenoceptor gene is considered to be a candidate gene in the development of BHR. Thus, ${\beta}_2$-adrenoceptor gene polymorphism may be associated with the BHR of adolescents with asthma remission, but not with the BHR of symptomatic asthma. To evaluate this hypothesis, ${\beta}_2$-adrenoceptor gene polymorphism at 2 sites (Arg16-Gly, Gln27-Glu) were examined. Methods : Two hundred two adolescents with BHR ($PC_{20}<18\;mg/mL$) and long term remission (neither asthma-related symptoms nor medication during the previous 2 years) of their asthma (remission group), 182 adolescents with symptomatic asthma (symptomatic group), and 200 healthy adolescents (control group) were studied. Asthma phenotypes were determined using methacholine bronchial provocation test and skin prick test. Genotypes of ${\beta}_2$-adrenoceptor polymorphism were evaluated by PCR-based methods. Results : Gly/Gly allele and Gly16-Gln27 haplotype were more prevalent in the remission group than in the control group (P=0.01, P=0.02), although there was no difference between the symptomatic group and the control group. In the remission group, there was significant difference in geometric mean of $PC_{20}$ among the 3 groups subdivided by the number of Gly16-Gln27 haplotype, showing that the Gly16-Gln27 haplotype was positively associated with BHR. However, no association was found between Gly16-Gln27 haplotype and BHR in the symptomatic group. Conclusion : This study demonstrates that ${\beta}_2$-adrenoceptor polymorphism at amino acid 16 and 27 was associated with BHR persisting in adolescents with asthma remission.
Ji-Hoon, Im;Minji, Oh;Youn-Lee, Oh;Min-Sik, Kim;Jong-Won, Lee
Journal of Mushroom
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v.20
no.4
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pp.218-226
/
2022
Flammulina velutipes, known as winter mushroom in the family of Physalacriaceae, is the main edible and export mushroom with the third highest production after oyster and king oyster mushroom in Korea. However, as normal consumers regard F. velutipes as a simple subsidiary material, there is a limitation to increasing mushroom demand. In order to overcome the consumption limit and increase the differentiation of new varieties, it is necessary to breed varieties with enhanced functionality in consideration of consumer preferences. Therefore, the study was performed to analyze nutrient components and several useful functional substances with 26 genetic resources of F. velutipes. Analyses of inorganic compound(Ca, K, Mg) and 15 amino acids revealed that Strain 4148 had the highest content among the 26 strains. Beta-glucan, which increases immune activity and polyphenol, which exert antioxidant effects were higher in non-white strains than in white strains with a small number of exceptions. Among the five fatty acids, linoleic acid(an omega-6 fatty acid) and α-linolenic acid(an omega-3 fatty acid), were detected in six mushroom strains. α-linolenic acid, which was not found in five major mushrooms including oyster mushrooms, was identified in F. velutipes. The results of HPLC analysis showed that 'Auram' (Strain 4232) and 'Baekseung'(Strain 4230) had the highest content of the stabilizing neurotransmitter GABA(15.38 ㎍/ml and 20.56 ㎍/ml, respectively) among non-white and white strains, respectively. Our findings provide useful information for breeding F. velutipes to obtain strains with enhanced functionality.
Two new analogs of bestatin were prepared from $\small{D}$-leucine and $\small{D}$-valine in a stereoselective and efficient way. An aminopeptidase inhibitor bestatin shows significant biological effects on immunomodulation and is marketed for the treatment of acute myelocytic leukemia. The key intermediates, trans-oxazolidine methyl esters 2a and 2b, were obtained with more than 20 to 1 stereoselectivity in a one-pot procedure by the three cascade reactions between N-hydroxymethyl protected ${\alpha}$-amino aldehydes (4a and 4b) and phenylsulfonylnitromethane ($PhSO_2CH_2NO_2$) and the following in-situ ozonolysis. Basic hydrolysis of 2a and 2b, and then the peptide coupling with $\small{L}$-Leu-OMe produced the protected derivatives of two new bestatin analogs, 3a and 3b, respectively. The new isobutyl and isopropyl analogs of bestatin (1a and 1b) were produced in overall 51% and 38% yields, respectively, with high stereoselectivity from the corresponding protected ${\alpha}$-amino aldehydes 4 in a six-step process.
Journal of the Korean Society of Food Science and Nutrition
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v.40
no.1
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pp.94-101
/
2011
Squid (Todarodes pacificus) is processed as dried or seasoned-dried products and its catch gradually increased from 270,298 M/T in 2005 to 367,940 M/T in 2008 in Korea. Squid processing by-product (viscera) was usually discarded as a waste resulting in environmental problem. In order to utilize squid viscera for more value-added products, a natural squid seasoning was developed by fermenting with Aspergillus oryzae koji. Squid viscera at 5, 10 and 15% salt concentrations with fixed levels of 5% koji and 30% water was fermented at room temperature. The quality properties of squid fermented products such as amino-N, TMA, VBN, total viable cell count, pH and total acidity were determined at different fermentation periods. The contents of amino-N, TMA, and VBN of squid seasoning at 5% salt concentration fermented for 14 days were the highest. Based on amino-N content, squid viscera at 5% koji fermented for 14 days was selected for further assays: the content of moisture, crude protein, crude lipid, crude ash, and carbohydrate were 5.98, 35.19, 33.08, 11.30, and 14.45%, respectively. The content of glutamate, alanine, leusine and lysine were 7.06, 12.34, 9.90 and 10.22%, respectively. The $IC_{50}$ values of DPPH scavenging and $\beta$-glucuronidase inhibitory activity were 12.89 and 12.58 mg/mL, respectively. A natural squid seasoning was manufactured by mixing fermented squid viscera and an ingredient. Based on the results of sensory evaluation, the fermented squid viscera seasoning was almost equal to other natural complex seasonings such as anchovy, cow meat, and fisheries seasoning.
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