• Asian-Australasian Journal of Animal Sciences
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• v.32 no.9
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• pp.1414-1422
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• 2019

Objective: Two experiments were conducted to determine the effects of adaptation time and inclusion level of sugar beet pulp (SBP) on nutrient digestibility and to evaluate the ileal amino acid digestibility of SBP fed to pigs. Methods: In Exp. 1, thirty-six crossbred barrows ($85.0{\pm}2.1kg$) were allotted to 6 diets in a completely randomized design with six replicates per diet. Diets included a corn-soybean meal diet and 5 test diets containing 14.6%, 24.4%, 34.2%, 43.9%, or 53.7% SBP, respectively. The adaptation time consisted 7, 14, 21, or 28 d consecutively for each pig followed by 5 d for fecal collection. Feces were collected from d 8 to 13, d 15 to 20, d 22 to 27, and d 29 to 34, respectively. In Exp. 2, six pigs ($35.1{\pm}1.7kg$) with T-cannulas at the terminal ileum were fed to 3 diets in a replicated $3{\times}3$ Latin square design with 3 periods and 2 replicate pigs per diet. Each period consisted 5 d for diet adaptation followed by 2 d for digesta collection. Results: The digestible energy (DE) value and the apparent total tract digestibility (ATTD) of gross energy (GE), dry matter (DM), ash, and organic matter in diets linearly decreased (p<0.05) as the adaptation time increased or as the dietary SBP increased, while the ATTD of neutral detergent fibre and acid detergent fibre in diets linearly increased (p<0.01) as the dietary SBP increased. The DE value and the ATTD of GE and crude protein (CP) in SBP linearly increased (p<0.05) as the adaptation time increased, while the ATTD of CP in SBP linearly decreased (p<0.01) as the inclusion level increased. The standardized ileal digestibility of Lys, Met, Thr, and Trp in SBP was 37.03%, 51.62%, 40.68%, and 46.22%, respectively. Conclusion: The results of this study indicated that the ATTD of energy and nutrients were decreased as inclusion rate of SBP increased.

• BMB Reports
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• v.56 no.6
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• pp.359-364
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• 2023

KAI1/CD82, a membrane tetraspanin protein, can prevent various cancers and retinal disorders through its anti-angiogenic and anti-metastatic capacity. However, little is known about its anti-inflammatory effect and molecular mechanism. Therefore, the present study aimed to inLPSvestigate effect of a recombinant protein of the large extracellular domain of human KAI1 (Gly 111-Leu 228, rhKAI1) on lipopolysaccharides (LPS)-stimulated RAW264.7 macrophage-like cells and mouse bone marrow-derived macrophages (BMDM) and to identify its underlying mechanism. Our data showed that rhKAI1 suppressed expression levels of classically macrophages (M1) phenotype-related surface markers F4/80+CD86+ in LPS-stimulated BMDM and RAW264.7 cells. In addition, LPS markedly increased mRNA expression and release levels of pro-inflammatory cytokines and mediators such as interleukin (IL)-1β, IL-6, tumor necrosis factor-α, cyclooxygenase-2, nitric oxide and prostaglandin E2, whereas these increases were substantially down-regulated by rhKAI1. Furthermore, LPS strongly increased expression of NF-κB p65 in the nuclei and phosphorylation of ERK, JNK, and p38 MAPK. However, nuclear translocation of NF-κB p65 and phosphorylation of JNK were greatly reversed in the presence of rhKAI1. Especially, rhKAI1 markedly suppressed expression of toll-like receptor (TLR4) and prevented binding of LPS with TLR4 through molecular docking predict analysis. Importantly, Glu 214 of rhKAI1 residue strongly interacted with Lys 360 of TLR4 residue, with a binding distance of 2.9 Å. Taken together, these findings suggest that rhKAI1 has an anti-inflammatory effect on LPS-polarized macrophages by interacting with TLR4 and down-regulating the JNK/NF-κB signaling pathway.

• Journal of Animal Science and Technology
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• v.59 no.12
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• pp.28.1-28.8
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• 2017

Background: Major amino acids in pig diets are Lys, Met, Thr, and Trp, but little is known about the requirements for the other essential amino acids, especially on odorous compounds and microbial characteristics in feces of growing-finishing pigs. To this end, different levels of amino acid composition added to diets to investigate the effects of amino acid composition on microbial characteristics and odorous compounds concentration. Methods: A total eight (n = 8) barrows (Landrace ${\times}$ Yorkshire ${\times}$ Duroc) with an average bodyweight of $89.38{\pm}3.3kg$ were individually fed diets formulated by Korean Feeding Standards 2007 (old version) or 2012 (updated with ideal protein concept) in metabolism crates with two replication. After 15-day adaptation period, fresh faecal samples were collected directly from pigs every week for 4 weeks and analysed for total volatile fatty acids (VFA), phenols and indoles by using gas chromatography. The nitrogen was determined by Kjeldahl method. Bacterial communities were detected by using a 454 FLX titanium pyrosequencing system. Results: Level of VFA tended to be greater in 2012 than 2007 group. Among VFAs, 2012 group had greater (p < 0.05) level of short chain fatty acids (SCFA) than control.Concentration of odorous compounds in feces was also affected by amino acid composition in pig diet. Levels of ammonium and indoles tended to be higher in 2012 group when compared with 2007 group.Concentration of phenols, p-cresol, biochemical oxygen demand, and total Kjeldahl nitrogen, however, were lower (P < 0.05) in 2012 treatment group compare to 2007. The proportion of Firmicute phylum were decreased, while the Bacteriodetes phylum proportion increased and bacterial genera includingCoprococcus, Bacillus, and Bacteroides increased (p < 0.05) in 2012 compare to 2007 group. Conclusion: Results from our current study indicates that well balanced amino acid composition reduces odor by modulating the gut microbial community. Administration of pig diet formulated with the ideal protein concept may help improve gut fermentation as well as reduce the odor causing compounds in pig manure.

• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.5
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• pp.832-836
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• 1994

This study was carried out to investigate several kinds of characteristics of pork sausage prepared by different cooking temperature and time (60, 90, 120, 150, 180 minutes in $58^{\circ}C$ and 25, 40, 55, 70, 85 minutes in $65^{\circ}C$). In case of color, L(bright), a (red) and b(yellow) value were 64.60-65.26, 9.14-9.94 and 8.68-9.34 in $58^{\circ}C$, and 65.16-66.68, 8.78-9.62 and 7.66-8.36 in $63^{\circ}C$, respectively. Gel strength showed high when cooking time was 120, 250 and 180 minute in $58^{\circ}C$ and 40 minute in $65^{\circ}C$. Residual nitrite concentration showed higher $58^{\circ}C$ than $65^{\circ}C$ and decreased gradually as cooking time elevated in all cooking temperature. Total plate count in 58$^{\circ}C$ was higher than $65^{\circ}C$, was wholly $8.7{\times}10^2~3.5{\times}10^3$.In case of free amino acid content, Asp, Glu and Lys were high and Cys, Met and Tyr low and was not different with $58^{\circ}C$ and $65^{\circ}C$. The result of sensory evaluation was not different (p<0.05) with $58^{\circ}C\;and\;65^{\circ}C$.

• Applied Biological Chemistry
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• v.33 no.4
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• pp.325-329
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• 1990

This study was undertaken to determine thermodynamic characteristics of B. subtilis p-4 and B. licheniformis p-5 proteases isolated from fermented anchovy paste. $K_m$ values of two proteases for casein as a substrate were 0.38mM in p-4 protease and 0.18mM in p-5 protease, respectively. Denaturation constants($K_D$) of p-4 and p-5 proteases were $12.2{\times}10^{-5}/sec\;and\;19.0{\times}10^{-5}/sec\;at\;40^{\circ}C,\;and\;35.7{\times}10^{-5}/sec\;and\;46.3{\times}10^{-5}/sec\;at\;50^{\circ}C$, respectively. Activation energies($E_a$) of p-4 and p-S pmteases were 19.6 Kcal/mole and 15.2kcal/mole, respectively. Free energy of activation(${\Delta}G^*$), activation enthalpy(${\Delta}H^*$) and activation entropy(${\Delta}S^*$) at $40^{\circ}C$ were 23.21Kcal/mole, 18.98Kcal/mole and -13.50 eu, respectively for p-4 protease and 22.93Kcal/mo1e, 14.58Kcal/mole and -26.68 eu, respectively for p-5 protease. The major amino acids in p-4 protease(151 residues of amino acid) were Gly, Glu, Pro, Asp, Ser, Ala, Lys and Leu, while those in p-5 protease(247 residues of amino acid) were Gly, Glu, Asp, Ala and Leu. It may be concluded that heat denaturation of two proteases showed liner regression curve and p-5 protease was more sensitive to heat than p-4 protease.

• The Korean Journal of Mycology
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• v.15 no.1
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• pp.29-37
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• 1987

Cellulolytic mocrooganisms were isolated from the cellulose-cultural properties. Among them, Aspergillus clavatus with the highest cellulase activity was identified by the morphological characteristics and it's enzyme activities were compared on the various cultural conditions. It was found that the induction of carboxymethylcellulase(CMCase), avicelase and salicinase in CMC liquid media showed the highest enzyme acitivity on five day's cultivation at $30^{\circ}C$ and thereafter their activities were gradually decreased with time. After crude extracellular enzymes precipitated with the 70% saturated ammonium sulfate solution were dialyzed with 20 mM acetate buffer pH 6.0, each crude enzyme was examined. The optimal activities of CMCase and avicelase were both found to be at $50^{\circ}C$ and pH 6.0. Their thermal stability was appeared at the $50^{\circ}C$. CMCase and avicelase had the maxima activities with 1.5% and 2.2% substrate concentration, respectively. The concentration of 5 mM $Mg^{++}$ or $Ca^{++}$ was found to have a maximum cellulase activity and its activity was inhibited with more than 5 mM $Cu^{++}$ and $Zn^{++}$ concentration. Cellulase activity was also inhibited sensitively by the inhibitors such as 2-mercaptoethanol, iodine and sodium azide.

• Applied Biological Chemistry
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• v.30 no.4
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• pp.345-350
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• 1987

Three lots of mejues were prepared with three different strains of Aspergillus oryzae, Bacillus natto, Bacillus subtilis and one was made by conventional method. The four different soy bean pastes were analyzed for compositions of free amino acids, sugars and organic acids during the period of fermentation. Amino nitrogen contents in the samples of A. oryzae were higher than others throughout the aging period. The amounts of each amino acids were varied markedly among the samples after 20-days, while glutamic acid, aspartic acid, lysine and phenylalanine were dominant in all samples after 90-days. Glucose contents were found to be in the range of $0.46{\sim}2.66%$ and other sugars of fructose, sucrose, rhamnose and maltose were less than 0.35%. The levels of total free sugar were relatively higher in the samples prepared with B. natto than others. Citric, lactic, malic, acetic and oxalic acids were identified, and the content of lactic acid was higher in the samples of A. oryzae, whereas citric acid was higher in conventional method.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.8
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• pp.1167-1173
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• 2009

This study was conducted to evaluate the effects of providing fermented soy protein to weaned pigs on nitrogen balance and apparent fecal and apparent ileal digestibility (AID) of AA. Four weaned ((Yorkshire${\times}$Landrace)${\times}$Duroc) barrows (BW = 6.58${\pm}$0.98 kg), surgically fitted with a simple T-cannula approximately 15 cm prior to the ileo-cecal junction, were fed four diets according to 4${\times}$4 Latin square design. Diets were a basal diet supplemented with one of the following: 3% SDPP (spray dried plasma protein), 5% RBP (soy protein fermented by Lactobacillus spp.), 5% PSP (soy protein fermented by Aspergillus oryzae and Bacillus subtilis), and 2.5% RPP (2.5% RBP+2.5% PSP). No differences were observed in DM and N intakes among treatments. However, the level of urine excretion was greater in the RPP group than in the PSP group. Additionally, fecal DM excretion, fecal N concentration and fecal N excretion were increased in the RBP, PSP and RPP groups when compared with the SDPP group (p<0.05). Furthermore, total excretion was increased in the RPP group when compared with the PSP group (p<0.05). In addition, N absorption and the N absorption ratio were higher in the SDPP group than in the RPP group (p<0.05). Moreover, the DM and N digestibilities were lower in the RBP, PSP and RPP groups than in the SDPP group (p<0.05), and the ash and energy digestibilities were higher in the SDPP and RBP groups than in the PSP and RPP groups (p<0.05). However, no significant differences were observed in the DM, N, Ash, Ca, P or ileal digestibilities among treatments, although the energy digestibility was higher in the SDPP group than the RBP group (p<0.05). In addition, the apparent ileal digestibilities of essential amino acids (Arg, His, Iso, Leu, Lys, Phe, Thr, and Val) were significantly higher in the SDPP group than in the other groups (p<0.05), and the levels of Ala, Cys, Glu and Try were greater in the SDPP treatment group than the RBP, PSP and RPP groups (p<0.05). Additionally, the levels of Asp, Gly and Ser were higher in the SDPP group than the PSP and RPP groups, and the level of Pro was higher in the SDPP group than the RPP group (p<0.05). Finally, total non-essential amino acid and total amino acid digestibility were higher in the SDPP group than in the other treatments (p<0.05). Taken together, the results of this study indicate that animal protein is more bioavailable than plant protein. However, the N absorption ratio and ileal digestibility were found to be similar in the SDPP and RBP groups.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.36-36
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• 2017

Out of twenty common protein amino acids, there are many kinds of non protein amino acids (NPAAs) that exist as secondary metabolites and exert ecological functions in plants. Mimosine (Mim), one of those NPAAs derived from L. leucocephala acts as an iron chelator and reversely block mammalian cell cycle at G1/S phases. 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 biosynthesis includes consecutive two steps using two enzymes-serine acetyl transferase (SAT) and O-acetylserine (thiol)lyase (OASTL) and appeared in plant cytosol, chloroplast, and mitochondria. In the first step, the acetylation of the ${\beta}$-hydroxyl of L-serine by acetyl-CoA in the existence of SAT and finally, OASTL triggers ${\alpha}$, ${\beta}$-elimination of acetate from OAS and bind $H_2S$ to catalyze the synthesis of Cys. Mimosine synthase, one of the isozymes of the OASTLs, is able to synthesize Mim with 3-hydroxy-4-pyridone (3H4P) instead of $H_2S$ for Cys in the last step. Thus, the aim of this study was to clone and characterize the cytosolic (Cy) OASTL gene from L. leucocephala, express the recombinant OASTL in Escherichia coli, purify it, do enzyme kinetic analysis, perform docking dynamics simulation analysis between the receptor and the ligands and compare its performance between Cys and Mim synthesis. Cy-OASTL was obtained through both directional degenerate primers corresponding to conserved amino acid region among plant Cys synthase family and the purified protein was 34.3KDa. After cleaving the GST-tag, Cy-OASTL was observed to form mimosine with 3H4P and OAS. The optimum Cys and Mim reaction pH and temperature were 7.5 and $40^{\circ}C$, and 8.0 and $35^{\circ}C$ respectively. Michaelis constant (Km) values of OAS from Cys were higher than the OAS from Mim. Inter fragment interaction energy (IFIE) of substrate OAS-Cy-OASTL complex model showed that Lys, Thr81, Thr77 and Gln150 demonstrated higher attraction force for Cys but 3H4P-mimosine synthase-OAS intermediate complex showed that Gly230, Tyr227, Ala231, Gly228 and Gly232 might provide higher attraction energy for the Mim. It may be concluded that Cy-OASTL demonstrates a dual role in biosynthesis both Cys and Mim and extending the knowledge on the biochemical regulatory mechanism of mimosine and cysteine.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.3
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• pp.230-237
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• 2018

This study investigated the food and nutritional characteristics of alkaline-insoluble fractions (AIFs) recovered from bastard halibut Paralichthys olivaceus (BH) and skipjack tuna Katsuwonus pelamis (ST) roes using the alkaline solubilization. The moisture content of AIFs ranged from 4.8% to 12.8%, and ST provided significantly better yields (9.5 for STAIF-11 and 7.1 g/100 g roe for STAIF-12) than did BH (P<0.05). The protein content of AIFs ranged from 71.7% to 79.2%, with the highest level yielded by STAIF-11 (6.8 g/100 g roe). The crude fat content of AIFs was 10.9-14.3% and the mineral content was 0.7-3.4%. The major mineral components of AIFs were sulfur, sodium, potassium, and phosphorus. Color values showed that BHAIFs were significantly brighter than STAIFs. Total contents of essential amino acids were significantly higher in STAIFs (47.5-49.5%) than in BHAIFs. The major essential amino acids found in AIFs from both sources were Val, Leu, Lys, and Arg. Therefore, AIFs were significantly superior to whole BH roe in terms of physicochemical and nutritional status, and we identified species-specific differences between BH and ST. Protein is a major component of AIFs recovered from fish roes, which suggests that they have potential for use as a protein source.