• Title/Summary/Keyword: L-lysine production

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Recombinant production of human glucagon-like peptide-1 mutant (인간 Glucagon-like Peptide-1 변이체의 재조합 생산)

  • Kim, Sung-Gun;Park, Jong-Tae
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.237-243
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    • 2014
  • Human Glucagon like peptide-1 (GLP-1) is an incretin hormone that promotes secretion of insulin. In order to eliminate the formation of the soluble aggregate, Ala19 in GLP-1 was substituted with Thr, resulting in a GLP-1 mutant GLP-1A19T. The gene synthesis of GLP-1A19T and the fusion of 6-lysine tagged ubiquitin gene were accomplished by using the overlap extension polymerase chain reaction. The ubiquitin fused GLP-1A19T (K6UbGLP-1A19T) is expressed as form of inclusion body with little formation of the soluble aggregation in recombinant E. coli. In order to produce K6UbGLP-1A19T in large amounts, fed-batch fermentation was carried out in a pH-stat feeding strategy. Maximum dry cell weight of 87.7 g/L and 20.4% of specific K6UbGLP-1A19T content were obtained. Solid-phase refolding using a cation exchanger was carried out to renature K6UbGLP-1A19T. The refolded K6UbGLP-1A19T aggregated little and was released GLP-1A19T by on-column cleavage with ubiquitin-specific protease-1. The molecular mass of GLP-1A19T showed an accurate agreement with its theoretical molecular mass.

Mass-Based Metabolomic Analysis of Lactobacillus sakei and Its Growth Media at Different Growth Phases

  • Lee, Sang Bong;Rhee, Young Kyoung;Gu, Eun-Ji;Kim, Dong-Wook;Jang, Gwang-Ju;Song, Seong-Hwa;Lee, Jae-In;Kim, Bo-Min;Lee, Hyeon-Jeong;Hong, Hee-Do;Cho, Chang-Won;Kim, Hyun-Jin
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.925-932
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    • 2017
  • Changes in the metabolite profiles of Lactobacillus sakei and its growth media, based on different culture times (0, 6, 12, and 24 h), were investigated using gas chromatography-mass spectrometry (MS) and liquid chromatography-MS with partial least squares discriminant analysis, in order to understand the growth characteristics of this organism. Cell and media samples of L. sakei were significantly separated on PLS-DA score plots. Cell and media metabolites, including sugars, amino acids, and organic acids, were identified as major metabolites contributing to the difference among samples. The alteration of cell and media metabolites during cell growth was strongly associated with energy production. Glucose, fructose, carnitine, tryptophan, and malic acid in the growth media were used as primary energy sources during the initial growth stage, but after the exhaustion of these energy sources, L. sakei could utilize other sources such as trehalose, citric acid, and lysine in the cell. The change in the levels of these energy sources was inversely similar to the energy production, especially ATP. Based on these identified metabolites, the metabolomic pathway associated with energy production through lactic acid fermentation was proposed. Although further studies are required, these results suggest that MS-based metabolomic analysis might be a useful tool for understanding the growth characteristics of L. sakei, the most important bacterium associated with meat and vegetable fermentation, during growth.

Effect of Daily Protein Allowances during Lay on Egg Production and Feed Cost of Broiler Breeders (육용종계 산란기의 단백질 공급수준이 산란능력과 경제성에 미치는 영향)

  • 함영훈;김상인;이규호
    • Korean Journal of Poultry Science
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    • v.30 no.2
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    • pp.101-106
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    • 2003
  • In order to study the optimum daily protein allowance for broiler breeders of 24 to 64 weeks of age on a control-fed basis, an experiment was carried out with 400 hens of Arbor Acres strain. Four levels of protein allowances were used to supply 18 to 24g of protein per day in 2g increments. As the age of hens increased, a same stepwise increase and decrease in daily energy allotment was used in all treatments. Same amount of calcium, phosphorus, methionine and lysine were supplied in all treatments and throughout laying period. Hen-day egg production was highest in hens receiving 20g protein per day(p<0.05), but there was no significant difference among those fed daily protein ranging 18 to 22g, and increasing the daily protein allotment up to 24g resulted in a significant decrease(p<0.05). Average egg weight showed a trend to increase as the daily protein allowance increase(p<0.05), but no significant difference was found among the hens fed daily protein 20 to 24g. Feed and ME conversion was superior in hens receiving 20g daily protein(p<0.05). CP conversion was increased as the daily protein allowance increase(p<0.05), but there was no significant difference between hens receiving 18 and 20g daily protein. Feed cost required per egg or per kg egg was lowest in hens fed 20g daily protein. It could be concluded that the optimum daily protein allowance was 20g in all performances.

Effects of Conditioners (Standard, Long-Term and Expander) on Pellet Quality and Growth Performance in Nursery and Finishing Pigs

  • Johnston, S.L.;Hines, R.H.;Hancock, J.D.;Behnke, K.C.;Traylor, S.L.;Chae, B.J.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.4
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    • pp.558-564
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    • 1999
  • A series of experiments were conducted to determine the effects of standard (ST), long-term (LT), and expander (EX) conditioners on nutritional value of phase-three nursery and finishing swine diets. In Exp. 1, 180 pigs (average initial BW of 11.7 kg) were fed com-soybean meal based diets (1.3% lysine) during a 28 d growth assay. Gain/feed was improved (p<0.004) with pelleting and pellet durability index (PDI) increased with degree of conditioning (LT>ST). However, there was no advantage for LT vs ST conditioning in rate or efficiency of gain (p>0.5). In Exp. 2, 180 pigs (average initial BW of 10.4 kg) were fed con-soybean meal based diets (0.9% lysine) during a 28 d growth assay, Pelleted diets tended to support greater ADG (p<0.08) and gain/feed (p<0.002) with no marked advantage from EX vs ST conditioning. In Exp 3, a total of 70 barrows (average initial BW of 54 kg) was used in a growth assay to determine the effects of feeding a com-soybean meal based diet processed with a standard (ST) steam conditioner, a long-term (LT) steam conditioner, and an expander (EX) conditioner. The conditioned diets were fed as mash (M) or pellets (P) to give a $2{\times}3$ factorial plus a meal control. PDI increased with degree of conditioning (EX>LT>ST). There was a trend (p<0.07) for greater ADG in pigs fed diets that had been thermally conditioned. Also, there was a general advantage in gain/feed with pelleting (p<0.04), but this advantage was pronounced only with standard conditioning. Indeed, the greatest gain/feed was observed for pigs fed the expander treatments (p<0.03) and the expander mash was used as efficiently as the expander pellets. There was no difference in backfat thickness among pigs fed the treatments (p>0.3). but the more extreme the processing technique. the greater the incidence and severity of stomach lesions (p<0.04). These results suggest maximum rate and efficiency of growth with pelleting after standard steam conditioning or simply feeding an expanded mash.

Physico-chemical properties of late-incubation egg amniotic fluid and a potential in ovo feed supplement

  • Omede, A.A.;Bhuiyan, M.M.;lslam, A.F.;Iji, P.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1124-1134
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    • 2017
  • Objective: This study explored the physico-chemical properties of late-incubation egg amniotic fluid and a potential in ovo feed (IOF) supplement. Methods: Amniotic fluid was collected from broiler breeders (Ross 308, 51 weeks and Cobb 500, 35 weeks) on day 17 after incubation. A mixture of high-quality soy protein supplement - Hamlet Protein AviStart (HPA) was serially diluted in MilliQ water to obtain solutions ranging from 150 to 9.375 mg/mL. The mixtures were heat-treated (0, 30, 60 minutes) in a waterbath ($80^{\circ}C$) and then centrifuged to obtain supernatants. The amniotic fluid and HPA supernatants were analysed for their physico-chemical properties. Results: Only viscosity and $K^+$ were significantly (p<0.05) different in both strains. Of all essential amino acids, leucine and lysine were in the highest concentration in both strains. The osmolality, viscosity and $pCO_2$ of the supernatants decreased (p<0.05) with decreasing HPA concentration. Heat treatment significantly (p<0.05) affected osmolality, pH, and $pCO_2$, of the supernatants. The interactions between HPA concentration and heat treatment were significant with regards to osmolality (p<0.01), pH (p<0.01), $pCO_2$ (p<0.05), glucose (p<0.05), lactate (p<0.01) and acid-base status (p<0.01) of HPA solutions. The $Ca^{2+}$, $K^+$, glucose, and lactate increased with increasing concentration of HPA solution. The protein content of HPA solutions decreased (p<0.05) with reduced HPA solution concentrations. The supernatant from 150 mg/mL HPA solution was richest in glutamic acid, aspartic acid, arginine and lysine. Amino acids concentrations were reduced (p<0.05) with each serial dilution but increased with longer heating. Conclusion: The values obtained in the primary solution (highest concentration) are close to the profiles of high-protein ingredients. This supplement, as a solution, hence, may be suitable for use as an IOF supplement and should be tested for this potential.

Effects of Sodium Sulfite and Extrusion on the Nutritional Value of Soybean Products for Nursery Pigs

  • Burnham, L.L.;Kim, I.H.;Kang, J.O.;Rhee, H.W.;Hancock, J.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.11
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    • pp.1584-1592
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    • 2000
  • Three hundred nursery pigs were used in two growth assays (avg initial BW of 6.5 and 6.0 kg, respectively) to determine the effects of sodium sulfite ($Na_2SO_3$) as an extrusion aid for soy products used in diets for weanling pigs. In Exp. 1, treatments were arranged as $3{\times}2$ factorial with main effects of soy product [soybean meal (SBM), extruded SBM, and dry-extruded whole soybeans (DEWS)] and concentration of $Na_2SO_3$ (0 and 10 g/kg of soy product). The extruded SBM and DEWS treatments were processed in a dry extruder ($Insta-Pro^{(R)} $, Triple F Nutrition, Des Moines, IA) with barrel temperatures and throughputs of $169^{\circ}C$ and 578 kg/h, and $147^{\circ}C$ and 598 kg/h, respectively. All diets were formulated to 3.5 Mcal/kg DE, with 0.92% lysine for d 0 to 14, and 0.76% lysine for d 14 to 28. For d 0 to 14, there was a tendency for pigs fed diets with $Na_2SO_3$ to have greater ADG (p<0.08), and pigs fed SBM to have greater ADFI (p<0.02), thus pigs fed the extruded soy products has 15% greater gain/feed than those fed SBM (p<0.007). For d 14 to 28, there were no differences in ADG or gain/feed among pigs fed diets with SBM and those fed diets with the extruded soy products (p>0.15). However, pigs fed DEWS had greater ADG than pigs fed extruded SBM, and pigs fed $Na_2SO_3$ had greater ADG and ADFI compared to those not fed $Na_2SO_3$ (p<0.02 and 0.08, respectively). The positive response in ADG and gain/feed to the addition of $Na_2SO_3$ resulted with SBM and extruded SBM treatments, and not with DEWS (interaction effect, p<0.04). Overall (d 0 to 28), pigs fed DEWS had greater ADG (p<0.01) and gain/feed (p<0.08) than pigs fed extruded SBM. Also pigs fed diets with $Na_2SO_3$ had greater ADG, ADFI, and gain/feed compared to those fed diets without $Na_2SO_3$ (p<0.002, 0.04, and 0.04, respectively). Exp. 2 was designed as a $2{\times}3$ factorial with main effects of soy product (SBM and DEWS) and concentration of $Na_2SO_3$ (none, 7.5, and 15.0 g/kg of soy product). As in Exp. 1, all diets were formulated to 3.5 Mcal/kg DE, with 0.92% lysine for d 0 to 13, and 0.76% lysine for d 13 to 53. At a constant processing temperature (148 to $149^{\circ}C$, $Na_2SO_3$ increased throughput of the extruder (578, 595, and 602 kg/h for the 0, 7.5, and 15.0 g/kg additions, respectively). For d 0 to 13, treatment had no effect on ADG or ADFI, but gain/feed decreased for pigs fed SBM with increasing concentrations of $Na_2SO_3$, and increased for pigs fed DEWS with increasing concentrations of $Na_2SO_3$ (SBM vs DEWS sulfite quadratic interaction, p<0.03). For d 13 to 35, pigs fed DEWS had greater ADG (p<0.01) and gain/feed (p<0.001) than pigs fed SBM. Also, ADFI decreased and gain/feed increased with increasing concentrations of $Na_2SO_3$ (linear effects, p<0.04 and 0.01, respectively). Overall, pigs fed the diets with DEWS had greater ADG and gain/feed than pigs fed SBM (p<0.003 and 0.002, respectively), and $Na_2SO_3$ tended to decrease ADFI and increase gain/feed (linear effects, p<0.07 and 0.06, respectively). In conclusion, pigs fed DEWS had greater rate and efficiency of gain than pigs fed SBM. Also, adding $Na_2SO_3$ prior to extrusion increased yield and feed efficiency.

Effects of Soybean Small Peptides on Rumen Fermentation and on Intestinal and Total Tract Digestion of Luxi Yellow Cattle

  • Wang, W.J.;Yang, W.R.;Wang, Y.;Song, E.L.;Liu, X.M.;Wan, F.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.1
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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.

Effect of varying the amount of water added on the characteristics of mash fermented using modified Nuruk for distilled-Soju production (가수량 변화가 개량누룩으로 발효한 증류식 소주용 술덧의 특성에 미치는 영향)

  • Choi, Han-Seok;Kim, Eu-Gene;Kang, Ji-Eun;Choi, Ji-Ho;Yeo, Soo-Hwan;Jeong, Seok-Tae
    • Food Science and Preservation
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    • v.21 no.6
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    • pp.908-916
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    • 2014
  • This study was conducted to investigate the effects of varying the amount of water added on the characteristics of mash fermented using modified Nuruk for distilled-Soju production. As the amount of water added to the mash increased from 120 to 300%, the pH dropped from 4.6 to 4.2, resulting in reductions in the acidity from 6.0 to 5.2, in the amino acid level from 6.0 to 4.2, and in the soluble-solid content from 18.4 to $7.4^{\circ}Brix$. The alcohol concentration of the mash was highest at 17.6%, when 150% water was added, while the alcohol yield showed water-content-dependent increases of 59.7, 74.5, 80.8, 82.8, 89.4, and 90.6% with 120, 150, 180, 200, 250, and 300% water added, respectively. The values of the organic-acid content in the mash were 207.85, 222.38, 222.06, 204.56, 194.34, and 204.34 mg/100 mL, showing the highest values when 150 and 180% water was added. The total amino-acid content showed water-content-dependent decreases at 474.60, 317.32, 241.89, 244.51, 189.00, and 208.12 mg/100 mL, with arginine, alanine, glutamic acid, glycine, isoleucine, leucine, lysine, phenylalanine, proline, serine, tyrosine, and valine as the major components. The concentrations of isobutanol, isoamyl alcohol, 1-propanol, and 2-phenylalcohol were 154.88~182.62, 320.59~394.47, 91.50~170.91, and 108.93~144.26 ppm, respectively, while ethyl acetate, acetaldehyde, furfural, and butyric acid were also detected.

Effects of Low Crude Protein Diets Supplemented with Synthetic Amino Acids on Performance, Nutrient Utilization and Carcass Characteristics in Finishing Pigs Reared Using a Phase Feeding Regimen

  • Lee, J.H.;Kim, J.H.;Kim, J.D.;Kim, S.W.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.5
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    • pp.655-667
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    • 2001
  • This experiment was conducted to investigate the effect of feeding a low CP diet supplemented with synthetic amino acids on performance, nutrient utilization and carcass characteristics of finishing pigs fed under a three-phase feeding regimen. Ninety-six finishing pigs (Landrace$\times$Large White$\times$Duroc), $55.75kg{\pm}0.65$ of initial body weight, were blocked by weight and sex and allotted to four dietary treatments in a randomized block design. There were six pens per treatment and four pigs per pen. Pigs were fed a 16%-14%-12% CP (for phase I-II-III, respectively), sequence of diets. Dietary treatments were 1) Control, 2) Con+L (a sequence of diets reduced in CP by l percentage unit with lysine (L) supplementation, 3) Con+LMT (a sequence of diets reduced in CP by 2 percentage unit with LYS, methionine (MET) and threonine (THE) supplementation) and 4) Con+LMTT (a sequence of diets reduced in CP by 3 percentage unit with LYS, MET, THR and tryptophan (TRP) supplementation). The finishing period (55 to 105 kg) was divided into three phases (55 to 72 kg, 72 to 90 kg and 90 to 105 kg). Pigs fed either the control or Con+L diet grew faster (p<0.05) than pigs fed the Con-LMT or Con+LMTT diet. There was no difference in ADFI among dietary treatments. Phosphorus (P) digestibility was lowest in the control group and highest in the Con+LMTT group (p<0.05). Within each phase, no significant differences in dry matter (DM) and CP digestibilities were found. Although some amino acid digestibilities were affected by dietary treatments, digestibilities of essential amino acids (EAA), non-essential amino acids (NEAA) and total amino acid were not significantly influenced by dietary treatments. For the entire experiment periods, Con+L, Con+LMT and Con+LMTT treatments resulted in 13.4, 18.8 and 21.6% lower total N excretion compared with the control. Con+LMT and Con+LMTT treatments showed significantly lower BUN concentration compared with the control and Con+L treatment (p<0.05), but there was no significant difference in BUN concentration between pigs fed the control and Con+L treatment or between pigs fed Con+LMT and Con+LMTT treatments (p>0.05). Carcass length, backfat thickness and carcass grade were not significantly affected by dietary treatments (p>0.05). In conclusion, reducing dietary CP level by 1 percentage unit and supplementing only LYS at each phase could be a very beneficial feeding strategy for finishing pigs fed under a three phase feeding regimen in terms of both environmental and economical aspects.

Gelation of Rapeseed Protein Induced with Microbial Transglutaminase (미생물성 Transglutaminase에 의한 유채단백질의 겔화)

  • Hyun, Eun-Hee;Kang, Yeung-Joo
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1262-1267
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    • 1999
  • Optimum reaction conditions for gel formation of rapeseed, Brassica napus, protein catalyzed by microbial TGase(transglutaminase) were evaluated by measuring breaking strength and deformation of gel. The polymerization of the protein gel was ascertained by SDS-PAGE and content of GL crosslinking$[{\varepsilon}-({\gamma}-glutamyl)lysine]$. In the reaction between rapeseed protein and TGase at $45^{\circ}C$ for 60 min, the breaking strength and deformation of the gel was the maximum at the ratio of 1 : 40 of enzyme to substrate. 10%(w/v) of rapeseed protein concentrate was optimum for gel production. The maximum breaking strength and deformation was shown at $45^{\circ}C$ The breaking strength increased linearly up to 90 min of the reaction time and remained unchanged. The breaking strength and deformation by TGase treatment was pH dependent and pH 7 was optimum for 10% rapeseed protein solution. SDS-PAGE analysis indicated that new band of highmolecular polymers were formed by the enzyme reaction, with disappearing the original bands of rapeseed protein. According to HPLC analysis. the content of GL crosslinking was increased from 0 to $7.14\;{\mu}mol/g$ gel for 90 min of the reaction time.

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