• Animal Bioscience
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• 제35권2호
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• pp.260-271
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• 2022

Objective: We aimed to investigate the effect of the differing amino acid (AA) release dynamics of two protein sources on the growth performance, nitrogen deposition, plasma biochemical parameters, and muscle synthesis and degradation of piglets when included in their diets at normal and low concentrations. Methods: Forty-eight piglets (Duroc×Landrace×Large White) with initial body weight of 7.45±0.58 kg were assigned to six groups and fed one of 6 diets. The 6 dietary treatments were arranged by 3×2 factorial with 3 protein sources and 2 dietary protein levels. They are NCAS (a normal protein content with casein), NBlend (a normal protein content with blend of casein and corn gluten meal), NCGM (a normal protein content with corn gluten meal), LCAS (a low protein content with casein), LBlend (a low protein content with blend of casein and corn gluten meal), LCGM (a low protein content with corn gluten meal). The release dynamics of AA in these diets were determined by in vitro digestion. The digestibility, utilization and biological value of nitrogen in piglets were determined by micro Kjeldahl method. Plasma insulin was measured by enzyme-linked immunosorbent assay kits. The protein expression of mediators of muscle synthesis and degradation was determined by western blotting. Results: Although the consumption of a low-protein diet supplemented with crystalline AA was associated with greater nitrogen digestion and utilization (p<0.05), the final body weight, growth performance, nitrogen deposition, and phosphorylation of ribosomal protein S6 kinase 1 and eIF4E binding protein 1 in the muscle of pigs in the low-protein diet-fed groups were lower than those of the normal-protein diet-fed groups (p<0.05) because of the absence of non-essential AA. Because of the more balanced release of AA, the casein (CAS) and Blend-fed groups showed superior growth performance, final body weight and nitrogen deposition, and lower expression of muscle ring finger 1 and muscle atrophy F-box than the CGM-fed groups (p<0.05). Conclusion: We conclude that the balanced release of AA from CAS containing diets and mixed diets could reduce muscle degradation, favor nitrogen retention, % intake and improve growth performance in pigs consuming either a normal- or low-protein diet.

• Journal of Animal Science and Technology
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• 제46권1호
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• pp.91-96
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• 2004

유단백질에서 유래된 아미노산은 주로 L-이성체의 형태로 존재하나 일부 미생물 세포벽의 구성성분과 일부 항생물질내 D-amnio acid(DAA)가 존재하고 있는 것으로 보고 되고 있으며 이는 유제품의 품질저하와 연관된다. 따라서 이 연구는 품질저하와 연관된다. 따라서 이 연구는 내냉성미생물 P. fluorescens와 P. putida에 의한 DAA 생성에 관해 실험하였다. 상업용 혼합 유산균주인 ABT-C를 접종하여 발효시간대별로 free D-amino acid를 분석하였으며, 실험결과 전체적인 DAA 함량은 매우 낮은 수준이고, 특히 발효시간대별 변화가 크게 없어 유산균의 증식에 의한 DAA의 생성은 매우 미미한 것으로 나타났고, 또한 일부 L-amino acid의 경우 유산균의 생육에 이용되는데 DAA의 경우 발효시간대별 함량이 큰 변화가 없는 것으로 보아 유산균의 이용성 역시매우 낮은 것으로 생각된다. 원유에서 분리한 P. fluorescens와 P. putida를 접종하여 배양시간대별로 free D-amino acid를 분석하였으며 실험결과 두 처리구 모두 배양시간에 따라 DAA의 생성이 증가되는 경향을 나타내었으며, P. putida의 경우 P. fluorescens에 비해 D-Pro의 증가량이 상대적으로 높게 나타났으나 나머지 4종은 P. fluorescens에 의한 생성량이 더욱 높게 나타났고, 특히 유해성이 높은 것으로 보고된 D-Asp 및 D-Ala의 함량이 매우 높게 나타났다. 따라서 DAA는 내냉성미생물에 의한 품질저하 지표로서 이용가능성과 더불어 인체의 유해성에 대한 위험성을 배제하는 차원에서도 매우 중요하게 인식되어야 할 것으로 사료된다

• Animal cells and systems
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• 제12권3호
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• pp.149-155
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• 2008

We have isolated and characterized Agrius convolvuli cDNA encoding a c-type lysozyme. The cDNA sequence encodes a processed protein of 139 amino acid residues with 19 amino acid residues amino-terminal signal sequence and 120 amino acid residues mature sequence. The amino acid residues responsible for the catalytic activity and the binding of the substrate are conserved. Agrius lysozyme has a high identity to Manduca sexta. Recombinant A. convolvuli lysozyme was expressed in Escherichia coli BL21(DE3) pLysS cells for pGEX 4T-1 expression vector. Their optimal conditions for the fusion protein expression and purification were screened. Lysozyme gene amplified with primers ACLyz BamHI and ACLyz XhoI was ligated into the pGEX 4T-1 vector, which contained the glutathione S-transferase(GST) gene for fusion partner. The fusion protein was induced by IPTG and identified by SDS-PAGE analysis. Molecular weight of the fusion protein was estimated to be about 45 kDa. Recombinant lysozyme, fused to GST, was purified by glutathion-Sepharose 4B affinity chromatography. Western blot analysis of this protein revealed an immunoreactivity with the anti-Agrius lysozyme.

• Asian-Australasian Journal of Animal Sciences
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• 제15권3호
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• pp.406-409
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• 2002

The effect of refeeding with various single essential amino acids on the recovery of plasma insulin-like growth factor-I (IGF-I) concentration in fasted young chickens was examined. Young chickens (29 days of age) were divided into 15 experimental groups. Chickens in one group were fed on the commercial diet ad libitum for 4 days. The remaining 56 chickens in 14 experimental groups were fasted. After 2 days of fasting, 52 chicks in 13 fasted groups were refed with one of the following experimental diets for 2 days. Eleven experimental diets were protein-free diets supplemented with one of 11 essential amino acids (Arg, Gly, His, Ileu, Leu, Met, Phe, Lys, Thr, Trp, Val). The remaining 2 experimental diets were a protein-free diet containing 11 essential amino acids and a protein-free diet not supplemented with amino acids. Birds in the remaining fasted group continued to be fasted for 2 days. Fasting for 2 days markedly reduced plasma IGF-I concentration. When fasted chickens were refed the protein-free diet containing either Gly alone or all essential amino acids, plasma IGF-I concentration was recovered to the level similar to that of fed chickens. Protein-free diet alone, however, failed to restore the reduced IGF-I concentration in plasma. Body weight loss modulated by feeding with protein-free diets supplemented with various single essential amino acids was associated with changes in plasma IGF-I concentrations. We concluded that body weight loss by feeding with a protein-free diet was lower than that of fasted chickens and that body weight loss associated with the decrease in plasma IGF-I concentration was modulated by feeding with protein-free diets containing various single essential amino acids.

• Asian-Australasian Journal of Animal Sciences
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• 제28권12호
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• pp.1774-1783
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• 2015

Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type) milk lysozyme gene (YML), was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa) with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML) was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75) which was expressed in P. pastoris with expression vector $pPICZ{\alpha}A$ and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

• Asian-Australasian Journal of Animal Sciences
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• 제29권5호
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• pp.689-694
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• 2016

The objective of the current study was to evaluate the effect of supplementation of either L-methionine (L-Met) or DL-methionine (DL-Met) to diets of starter pigs on nitrogen (N) balance, metabolism, and serum amino acid profile. Eighteen crossbred ($Duroc{\times}Landrace{\times}Yorkshire$) barrows weighing $15.45{\pm}0.88kg$ were randomly allotted to 1 of 3 diets with 6 pigs per treatment. The diets included a basal diet (Met-deficient diet) containing 0.24% standardized ileal digestibility Met with all other essential nutrients meeting the pig's requirements. The other two diets were produced by supplementing the basal diet with 0.12% DL-Met or L-Met. The experiment lasted for 18 days, consisting of a 13-day adaptation period to the diets followed by a 5-day experimental period. Pigs were fed ad libitum and free access to water throughout the experiment. Results showed that the supplementation of either L-Met or DL-Met improved N retention, and serum methionine concentration, and decreased N excretion compared with basal diet (p<0.01). The N retention of pigs fed diets supplemented with the same inclusion levels of DL-Met or L-Met were not different (p>0.05). In conclusion, on equimolar basis DL-Met and L-Met are equally bioavailable as Met sources for starter pigs.

• Asian-Australasian Journal of Animal Sciences
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• 제9권3호
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• pp.309-313
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• 1996

This experiment was carried out to investigate the effect of rumen-protected lysine (RPLys) on growth rate, feed efficiency and plasma amino acid concentrations in sheep. RPLys was supplemented at the level of 0% ($T_1$), 0.2% ($T_2$) and 0.4% ($T_3$) of total DMI with 24 sheep in a 56 day feeding trial. The results are summarized as follows: 1. live weight gain of sheep in groups $T_1$, $T_2$ and $T_3$ was 219, 216 and 244 g/d, and was significantly (p < 0.05) higher for $T_3$ through the entire experiment. 2. Feed intake was not affected by RPLys supplementation. 3. The group fed $T_3$ had a significantly (p < 0.05) better feed efficiency than the groups fed $T_1$ and $T_3$. The response of $T_3$ was higher in growing period II of feeding low protein basal diet than in period I. 4. Plasma lysine concentrations tended to be higher with supplementing RPLys, but there were no differences between $T_2$ and $T_3$. 5. Supplementing RPLys in the diets increased plasma concentrations of arginine, asparagines, threonine, serine, valine and leucine compared with sheep receiving no RPLys. In contrast, plasma histidine was lower in sheep fed the supplementing RPLys than fed the diet $T_1$ with significant (p < 0.05) difference.

• Asian-Australasian Journal of Animal Sciences
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• 제26권1호
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• pp.72-81
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• 2013

Four Luxi beef cattle ($400{\pm}10$ kg) fitted with ruminal, duodenal and ileal cannulas were used in a $4{\times}4$ Latin square to assess the effects of soybean small peptide (SSP) infusion on rumen fermentation, diet digestion and flow of nutrient in the gastrointestinal tract. The ruminal infusion of SSP was 0 (control), 100, 200 and 300 g/d. Ruminal SSP infusion linearly (p<0.01) and quadratically (p<0.01) increased microbial protein synthesis and rumen ammonia-N concentration. Concentrations of total volatile fatty acid were linearly increased (p = 0.029) by infusion SSP. Rumen samples were obtained for analysis of microbial ecology by real-time PCR. Populations of rumen Butyrivibrio fibrisolvens, Streptococcus bovis, Ciliate protozoa, Ruminococcus flavefaciens, and Prevotella ruminicola were expressed as a proportion of total Rumen bacterial 16S ribosomal deoxyribonucleic acid (rDNA). Butyrivibrio fibrisolvens populations which related to total bacterial 16S rDNA were increased (p<0.05), while Streptococcus bovis populations were linearly (p = 0.049) and quadratically (p = 0.020) decreased by infusion of SSP. Apparent rumen digestibility of DM and NDF were (Q, p<0.05; L, p<0.05) increased with infusion SSP. Total tract digestion of DM, OM and NDF were linearly (p<0.01) and quadratically (p<0.01) increased by infusing SSP. The flow of total amino acids (AA), essential amino acids (EAA) and individual amino acids were linearly (p<0.01) and quadratically (p<0.01) increased with infusion SSP. The digestibility of Lysine was quadratically (p = 0.033) increased and apparent degradability of Arginine was linearly (p = 0.032) and quadratically (p = 0.042) increased with infusion SSP. The results indicated that infusion SSP could improve nutrient digestion, ruminal fermentation and AA availability.

• 한국초지조사료학회지
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• 제32권4호
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• pp.343-352
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• 2012

Sulphur deficiency has become widespread over the past several decades in most of the agricultural area. Oilseed rape (Brassica napus L.) is a very sensitive to S limitation which is becoming reduction of quality and productivity of forage. Few studies have assessed the sulphur mobilization in the source-sink relationship, very little is known about the regulatory mechanism in interaction between sulphur and nitrogen during the short-term sulphur deficiency. In this study, therefore, amount of sulphur and nitrogen incorporated into amino acids and proteins as affected by different S-supplied level (Control: 1 mM ${SO_4}^{2-}$, S-deficiency: 0.1 mM ${SO_4}^{2-}$, and S-deprivation: 0 mM ${SO_4}^{2-}$) were examined. The amount of sulphur in sulphate (S-sulphate) was significantly decreased by 25.8% in S-deprivation condition, compare to control, but not nitrogen in nitrate (N-nitrate). The markedly increase of sulphur and nitrogen incorporated amino acids (S-amino acids and N-amino acids) was observed in both S-deficiency and S-deprivation treatments. The amount of nitrogen incorporated proteins (N-protein) was strongly decreased as sulphur availability while the amount of sulphur incorporated into proteins (S-protein) was not affected. A highly significant ($p{\leq}0.001$) relationship between S-sulphate and S-amino acid was observed whereas the increase of N-amino acids is closely associated with decrease of N-proteins. These data indicate that increase of sulphur and nitrogen incorporated into amino acids was from different nitrogen and sulphur metabolites, respectively

• Journal of Animal Science and Technology
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• 제50권1호
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• pp.99-110
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• 2008

pH 조절법을 활용하여 닭가슴살을 이용한 수리미 제조 시 냉동변성방지제의 첨가 수준의 차이가 닭가슴살 수리미의 냉동 저장 중 수리미의 저장 특성에 미치는 효과에 대해 알아보고자, 대조구(Alaska pollack에 수세법을 활용하여 냉동변성방지제인 4% sugar, 5 % sorbitol 및 0.3% polyphosphate를 첨가)와, 닭가슴살을 이용하여 pH 조절법으로 제조된 처리구(T1: 5% sorbitol and 0.3% polyphosphate, T2: 4% sugar, 5% sorbitol and 0.3% polyphosphate, and T3: 2% salt, 4% sugar, 5% sorbitol and 0.3% poly- phosphate) 들로 나눠 실험에 공시하였다. 아미노산 조성에서 대조구가 처리구들보다 유의적으로 높게 나타났다. 또한 대조구는 저장기간이 증가할수록 아미노산 조성별 차이를 보이지 않았으나, 처리구들은 저장 30일째 아미노산 함량이 증가하는 것으로 나타났다. 지방산 조성의 경우, 대조구가 처리구에 비해 myristic acid, palmitic acid 및 palmitoleic acid 등이 높게 나타났으며, oleic acid, linoleic acid 및 arachidonic acid 함량은 낮게 나타났다(P<0.05). 그러나 대조구는 저장기간에 따른 함량의 변화가 없는 반면, 포화지방산 함량은 T2 및 T3에서 저장기간이 증가할수록 감소하였으며, 불포화지방산 함량은 증가하였다. 지방산패도는 모든 처리구에서 15일째까지는 감소하다가 저장 30일째에는 증가하였다. 처리 간에는 저장 30일째에 대조구와 T2에서 높게 나타났으며, T1 및 T3에서 낮은 지방산패도를 나타내었다(P<0.05). 특히 대조구 및 T2에서 높은 지방산패도를 나타내었다. 휘발성염기태질소화합물(VBN)의 경우, T1 및 T3는 저장기간이 증가할수록 높은 값을 보였으나, T2는 저장기간별 차이를 나타내지 않았으며, 대조구는 오히려 감소하여 저장 30일째에는 가장 낮은 값을 나타내었다. 특히 닭가슴살을 활용한 처리구들에 비해 대조구가 월등히 높은 VBN값을 나타내었다. 수분활성도의 경우 다른 처리구에 비해 대조구는 저장기간별 수분활성도에 차이를 보였으며, 처리 간에는 T1 및 T2가 대조구와 T3에 비해 저장 0일째에 높은 값을 나타내었다.총세균수는 모든 처리구에서 저장 15일째까지는 증가하다가 저장 30일째에는 감소하였으며, 처리 간에는 T1이 모든 저장기간에서 유의적으로 높은 총균수를 나타내었다. 살모넬라균의 경우 대조구는 저장기간이 증가할수록 감소하였으나, 처리구들은 저장 15일째까지는 증가하다가 30일째에는 감소하는 것으로 나타나 총균수와 유사한 경향을 보였다. 처리 간에는 대조구에 비해 T1 및 T2가 높은 균수를 나타내었다. 락토바실러스균은 저장 0일째에 T3에서 높은 값을 나타내었으나, 저장 15일 및 30일째 모든 처리구에서 검출되지 않았다. 또한 관능평가 결과, 색을 포함한 모든 관능평가 항목에서 대조구와 T3에서 T1과 T2에 비해 높은 선호도를 보였다. 또한 처리구별 비교 시 소금과 설탕을 첨가하지 않은 T1에 비해 첨가량이 증가할수록 높게 나타났다. 이상의 결과를 종합해보면, 처리구들에 비해 대조구가 아미노산 및 포화지방산 함량은 높게 나타났으며, 낮은 미생물 수를 나타내었다. 그러나 지방산패도 및 VBN 값은 높게 나타나 대조구의 저장성에 나쁜 영향을 미칠 것으로 판단된다. 또한 2% 소금 및 4% 설탕을 첨가한 T3에서 T1과 T2에 비해 높은 선호도를 보여 냉동변성방지제의 첨가가 수리미의 관능특성을 개선시킬 수 있을 것으로 판단된다.