• Title/Summary/Keyword: Protein digestion

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Expession of the Recombinant Klebsiella aerognes UreF Protein as a MalE Fusion

  • Kim, Keun-Young;Yang, Chae-Ha;Lee, Mann-Hyung
    • Archives of Pharmacal Research
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    • v.22 no.3
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    • pp.274-278
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    • 1999
  • Expression of the active urease of the enterobacterium, Klebsiella aerogens, requires the presence of the accessory genes (ureD, ureE, ureF, and ureG) in addition to the three structural genes (ureA, ureB, and ureC). These accessory genes are involved in functional assembly of the nickel-metallocenter for the enzyme. Characterization of ureF gene has been hindered, however, since the UreF protein is produced in only minute amount compared to other urease gene products. In order to overexpress the ureF gene, a recombinant pMAL-UreF plasmid was constructed from which the UreF was produced as a fusion with maltose-binding protein. The MBP-UreF fusion protein was purified by using an amylose-affinity column chromatography followed by an anion exchange column chromatography. Polyclonal antibodies raised against the fusion protein were purified and shown to specifically recognize both MBP and UreF peptides. The UreF protein was shown to be unstable when separated from MBP by digestion with factor Xa.

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In situ Ruminal and Intestinal Digestibility of Crude Protein and Amino Acids in By-product Feedstuffs

  • Baek, Youl Chang;Jeong, Jin young;Oh, Young Kyoon;Kim, Min Seok;Lee, Hyun jung;Jung, Hyun jung;Kim, Do hyung;Choi, Hyuck
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.38 no.2
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    • pp.74-83
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    • 2018
  • The objectives of this study was to evaluate the degradability and digestibility of crude protein (CP), rumen undegradable protein (RUP), and individual amino acids (AA) on six by-product feedstuffs (BPF) (rice bran, RB; wheat bran, WB; corn gluten feed, CGF; tofu residue, TR; spent mushroom substrate from Pleurotus ostreatus, SMSP; brewers grain, BG) as ruminants feed. Three Hanwoo steers (40 months old, $520{\pm}20.20kg$ of body weight) fitted with a permanent rumen cannula and T-shaped duodenal cannula were used to examine of the BPF using in situ nylon bag and mobile bag technique. The bran CGF (19.2%) and food-processing residue BG (19.7%) had the highest CP contents than other feeds. The RUP value of bran RB (39.7%) and food-processing residues SMSP (81.1%) were higher than other feeds. The intestinal digestion of CP was higher in bran RB (44.2%) and food-processing residues BG (40.5%) than other feeds. In addition, intestinal digestion of Met was higher in bran RB (55.7%) and food-processing residues BG (44.0%) than other feeds. Overall, these results suggest that RB and BG might be useful as main raw ingredients in feed for ruminants. Our results can be used as baseline data for ruminant ration formulation.

Application of Temperature Gradient Gel Electrophoresis To cAMP Receptor Protein (온도 기울기 전기영동장치의 CAMP 수용성 단백질에 응용)

  • Gang, Jong-Back;Cho, Hyun-Young
    • Journal of Life Science
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    • v.14 no.2
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    • pp.309-314
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    • 2004
  • Cyclic AMP receptor protein (CRP) is involved in the transcriptional regulation of more than 100 genes in E. coli. CRP dimer is converted into active form via the sequential conformation change of cAMP binding pocket, hinge region and HTH DNA binding motif by binding of cAMP. The temperature gradient gel electrophoresis (TGGE) was applied to CRP protein to know whether it was an efficient technique to study the conformational transitions and the thermal stability. TGGE showed the unfolding process of wild-type and S83G CRP proteins with the temperature gradient set from 29 to 71$^{\circ}C$ on nondenaturing polyacrylamide gel. Melting temperature (Tm) was 57$\pm$1 and 55$\pm$1$^{\circ}C$ for wild-type and S83G CRP, respectively in acidic buffer[89.8 mM Glycine and 24 mM Boric acid (pH 5.8)].

Characterization of 27K Zein as a Transmembrane Protein

  • Lee, Dong-Hee
    • BMB Reports
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    • v.31 no.2
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    • pp.196-200
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    • 1998
  • Zeins, maize storage proteins, are retained in the endoplasmic reticulum (ER) during the subcellular targeting process without the ER retention signal. Circumstantial data indicate that the 27K zein is an ER transmembrane protein. The potential transmembrane domain may permit the 27K zein to remain in the ER. This study investigated the potential transmembrane feature by employing alkaline extraction, proteinase K digestion, and surface biotinylation on isolated intact protein bodies. These assays consistently support the possibility of the 27K zein as a transmembrane protein. The 27K zein polypeptide was shown to be associated with alkali-stripped membranes. The polypeptide was digested by proteinase K to a smaller fragment. According to surface biotinylation, the 27K zeins was labeled to the exclusion of other classes of zeins. This study, therefore, concludes that the 27K zein has an ER transmembrane domain, which may serve as an anchor for zeins' ER retention.

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Studies on the Digestion of Beef by Ficin Treatment V. A Transmission Electron Microscopic Observation (Ficin 처리시 우육의 단백질 분해에 관한 연구 V. 투과형 전자현미경에 의한 관찰)

  • 김정숙
    • The Korean Journal of Food And Nutrition
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    • v.2 no.2
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    • pp.21-26
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    • 1989
  • The morphological changes of fresh beef treated with ficin(0.1% : 2 hrs, 6 hrs)were examined with transmission electron microscope(TEM), the results obtained were as follows ; Connective tissue protein in fresh beef treated with ficin became gradually fragmentation and was occurred solubilization with time The length of sarcomere in myofibrillar protein was elongated, M-line became dim, and the 1-band of Z-line was broken and beck me fragmentation with time.

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Studies on the Digestion of Beef by Ficin Treatment : III. A Scanning Electron Microscapic Observation (Ficin처리시 우육의 단백질 분해에 관한 연구 III. 주사형 전자 현미경에 의한 관찰)

  • 김정숙
    • The Korean Journal of Food And Nutrition
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    • v.1 no.2
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    • pp.1-8
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    • 1988
  • The morphological changes of fresh beef treated with ficin(0.1% : 35$^{\circ}C$ 2hrs, 6hr,) were examined with scanning electron microscope(SEM), the results obtained were as follows ; Connective tissue protein in fresh beef treated with ficin was occurred solubilization with time and gradually trasformed from a definite form into amorphous form, followed by showing an opening-up phenomenon again and subdivision. Myofibrillar protein was cracked and breaked slightly, followed subdivision with time.

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Studies on the Digestion of Beef by Ficin Treatment : IV. An Optical Microscopic Observation (Ficin처리시 우육의 단백질 분해에 관한 연구 IV. 광학 현미경에 의한 관찰)

  • 김정숙;주순재
    • The Korean Journal of Food And Nutrition
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    • v.1 no.2
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    • pp.9-17
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    • 1988
  • The morphological changes of fresh beef treated with ficin(0.1% : 2hrs, 6hrs) were examined with optical microscope (OM, LM), the results obtained were as follows : Connective tissue protein in fresh beef treated with ficin was occurred swelling and separation of endomysial reticulum with time, followed showing granulation and solubilization slightly. Myofibrillar protein was loosed wavy contractile muscle fiber, and showed erosin, cracks and breaks in fibers with time.

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Chemical Composition, In situ Digestion Kinetics and Feeding Value of Oat Grass (Avena sativa) Ensiled with Molasses for Nili-Ravi Buffaloes

  • Khan, Muhammad Ajmal;Sarwar, M.;Nisa, M.;Iqbal, Z.;Khan, M.S.;Lee, W.S.;Lee, H.J.;Kim, H.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.8
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    • pp.1127-1133
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    • 2006
  • This study examined the effect of cane molasses and fermentation time on chemical composition and characteristics of oat grass silage (OGS) and its in situ digestion kinetics, intake, digestibility, milk yield and composition in buffaloes (Bubalus bubalis). Oat grass (OG) harvested at 50-days of age was ensiled in laboratory silos with cane molasses at the rate of 0, 2, 4 and 6% of OG dry matter (DM) for 30, 35 and 40 days. Silage pH was decreased while lactic acid content increased with increasing level of cane molasses and fermentation time. Dry matter (DM), crude protein (CP) and true protein (TP) content of OGS were (p<0.05) significantly higher with higher cane molasses levels. However, they were not affected by the fermentation time. Similar trends were observed for neutral detergent fiber (NDF), acid detergent fiber (ADF), cellulose, acid detergent lignin and ash content of OGS. The OG ensiled for 30-days with 2% molasses was screened from laboratory study and used to determine comparative in situ DM and NDF digestion kinetics of OG and its silage. In situ DM and NDF digestibilities of OG were significantly (p<0.05) higher than OGS. Ruminal DM and NDF lag time, rate and extent of digestion of OG and its silage were similar. Two experimental diets of OG and OGS were formulated using 75:25 forage to concentrate ratio on a DM basis. Dry matter and CP intakes were similar in lactating buffaloes fed either OG- or OGS-based diets. However, NDF intake was higher in buffaloes fed the OG-compared with OGS-based diet. Apparent DM, CP and NDF digestibilities were similar in lactating buffaloes fed either OG- or OGS-based diets. Milk yield (4% FCM) was similar in buffaloes fed either OG-(10.3 kg/d) or OGS-(9.95 kg/d) based diets. Milk fat, total solids and true protein content were higher with OG compared with the OGS diet. Solids not fat and CP content were similar in milk of buffalo fed either OG or OGS. The results of this study indicate that OG ensiled with 2% molasses could safely replace 75% DM of green oat fodder in the diets of lactating buffaloes without negatively affecting intake, digestibility, milk yield and composition.

Influence of Varying Dry Matter and Molasses Levels on Berseem and Lucerne Silage Characteristics and Their In situ Digestion Kinetics in Nili Buffalo Bulls

  • Touqir, N.A.;Khan, M. Ajmal;Sarwar, M.;Nisa, M.;Lee, W.S.;Lee, H.J.;Kim, H.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.887-893
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    • 2007
  • Influences of forage DM and addition of cane molasses on silage characteristics of berseem (Trifolium alexandrium) and lucerne (Medicago sativa) and their ruminal digestion kinetics in Nili buffaloes were studied. Berseem and lucerne fodders (at one tenth bloom) were ensiled with wheat straw in laboratory silos to achieve 20, 30, 40% forage DM and without wheat straw (control); each forage DM level was supplemented with 2, 4 and 6% of cane molasses at ensiling. The pH and lactic acid contents of berseem and lucerne silages were affected by both forage DM and addition of molasses. Dry matter, CP and true protein (TP) of berseem and lucerne silages were affected by forage DM at ensiling but were not affected by the addition of cane molasses. Higher DM, CP and TP losses were observed when berseem and lucerne fodders were ensiled either without wheat straw or with wheat straw to achieve 20% and 40% forage DM at ensiling compared with 30% DM at ensiling. Fiber fractions (NDF, ADF, hemicellulose and cellulose) of berseem silage and lucerne silage were significantly increased with increasing forage DM at ensiling. Addition of cane molasses did not affect the DM, CP, TP and fiber fractions of both berseem and lucerne silages. Berseem and lucerne ensiled at 30% DM with 2% cane molasses were screened for comparative ruminal digestion kinetics with their respective fodders. Addition of wheat straw to berseem or lucerne fodder at ensiling depressed DM and NDF ruminal degradability. However, ruminal lag time, rate of degradation and extent of digestion of silages were similar to their respective fodders. In conclusion, berseem and lucerne could be ensiled with wheat straw to increase their DM to 30% along with 2% molasses for buffaloes.

Influence of Chicken Embryo Extract on Protein Synthesis of Chicken Embryo Myoblasts Depends on Cell Density

  • Kita, K.;Hiramatsu, K.;Okumura, Jun-ichi
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.6
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    • pp.713-717
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    • 1998
  • The synergistic effect of fetal calf serum (FCS) and chicken embryo extract (CEE) on protein synthesis of chicken embryo myoblasts was examined. Myoblasts were derived from chicken embryo cultured for 14 days by trypsin digestion and cultured in 5% $CO^2/95%$ air at $37^{\circ}C$. When myoblasts were cultured at the low level of cell density (20-50% of well), CEE enhanced the ability of FCS to stimulate protein synthesis of myoblasts. However, there was no significant effect of CEE to stimulate protein synthesis of myoblasts cultured at high level of cell density (100% of well).