• Asian-Australasian Journal of Animal Sciences
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• 제24권2호
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• pp.272-278
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• 2011

Bovine whey protein expression patterns of colostrum are much different from that of milk. Moreover, bovine colostrum is an important source of protective, nutritional and developmental factors for the newborn. However, to our knowledge, no research has been performed to date using a comparative proteomic method on the changes in the bovine whey proteome during the transition from colostrum to milk. This study therefore separated whey protein of days 1, 3, 7 and 21 after calving using two dimension electrophoresis. Differentially expressed proteins at different collection times were identified using high-performance liquid chromatography in tandem with mass spectrometry (LC/MS) and validated by enzyme-linked immunosorbent assay (ELISA) in order to understand the developmental changes in the bovine whey proteome during the transition from colostrum to milk. The expression patterns of whey protein of days 1 and 3 post-partum were similar except that immunoglobulin G was down-regulated on day 3, and four proteins were found to be down-regulated on days 7 and 21 compared with day 1 after delivering, including immunoglobulin G, immunoglobulin M, albumin, and lactotransferrin, which are involved in immunity and molecule transport. The results of this study confirm the comparative proteomic method has the advantage over other methods such as ELISA and immunoassays in that it can simultaneously detect more differentially expressed proteins. In addition, the difference in composition of milk indicates a need for adjustment of the colostrum feeding regimen to ensure a protective immunological status for newborn calves.

• 농업과학연구
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• 제45권4호
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• pp.635-643
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• 2018

Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.

• 농업과학연구
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• 제39권4호
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• pp.561-568
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• 2012

The aim of this study was to introduce a simple method for isolation of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin from cow's milk, and peptides produced by enzymatic hydrolysis of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Whey protein were precipitated from whey by ammonium sulfate and, ${\alpha}$-lactalbumin and ${\beta}$-lactoglobulin were isolated using Hi Prep 26/60 Sephacryl S-100 column gel filtration chromatography. Bovine serum albumin and ${\beta}$-lactoglobulin were isolated by Mono-Q 5/50 GL column anion exchange chromatography of the 50% Ammonium Sulfate-supernatant. Isolated whey proteins were hydrolyzed by proteolytic alcalase. Tricine SDS-PAGE and reverse-phase HPLC analyses revealed that almost hydrolyzed all the ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Molecular weight of various peptides derived from alcalase hydrolysate were small molecular weight than 3.5 kDa.

• 대한수의학회지
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• 제47권1호
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• pp.1-6
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• 2007

The importance of milk for the growth and health of a newborn offspring is well known. Milkcontains immunoglobulin G (Ig G), Ig A, lactoperoxidase, lactoferin, cytokines, and growth factors.Osteopontin, one of the multifunctional proteins, is secreted by macrophages, T cel, and epithelial cells.bovine milk have not been clarified. The aim of this study was to observe the expression of osteopontin,in bovine milk during the lactation period or bovine mamary glands..Western blot analysis detected thatosteopontin was expressed in bovine milk whey and mamary glands. The expression level of osteopontinin colostrum whey was higher than those in early milk and mature milk whey. Immunohistochemistryshowed that osteopontin was detected in the glandular epithelium and epithelial cels of intralobular ductof mamary glands. These findings suggest that osteopontin transiently shows high expression in colostrumand plays a potential role in the immunological development of breast-fed calves.

• 한국식품위생안전성학회지
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• 제5권4호
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• pp.219-228
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• 1990

생유와 시판되고 있는 저온살균유, 고온 순간살균유 및 초고온순간살균유 각 20예에 대하여 polyacrylamide 겔 전기 영동법으로 단백질의 조성을 구하였다. 생유의 Whey 단백질 중 혈청 albumin, ${\alpha}-lactalbumin,\;{\beta}-lactoglobulin$의 조성은 3.71 : 11.44 : 84.85였으며 저온 살균과 고온순간살균에 의하여 큰 변호를 주지 않았다. 그러나 초고온 순간 살균으로 조성비는 0 : 64.75 : 35.5로 변화되어 형청 albumin의 소실과 ${\beta}-lactoglobulin$의 감소를 보였다. 살균처리유의 냉장보존 3일째에 ${\alpha}-lactalumin과\;{\beta}-lactoglobulin$의 감소가 있었으나 $25^{\circ}C$ 실온보존에서는 변화가 없었다. 고온 순간 살균유는 실온보존 2일째에 혈청 albumin, 3일째에 ${\alpha}-Lactalbumin의$ 감소만 있었고 다른 Whey 단밸질은 보존온도에 관계없이 3일간 농도변화가 없었다. 초고온순간살균유는 냉장보존 2일째 혈청 albumin의 감소만 인정되었다.

• 한국축산식품학회지
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• 제25권2호
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• pp.250-256
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• 2005

To investigate the effect of bovine colostral whey fractions on in vitro proliferation of mouse splenocytes, polypeptide fractions were separated from acid whey into 3 fractions depending on molecular weight by ultrafiltration: Fraction I, which contains the polypeptide larger than 10,000 Da, Fraction n, which contains the polypeptide ranging from 1,000 Da to 10,000 Da and Fraction III, which contains the polypeptide smaller than 1,000 Da. Fraction II showed the highest proliferative effect of mouse splenocytes among the colostral whey fractions and this proliferative activity increased in dose dependent manner. Unheated Fraction II and Fraction III showed significantly (p<0.01) higher proliferative effects than others but heated Fraction II showed the highest enhancing effect of mouse splenocyte among heated whey fractions (p<0.01). The supplementation of Fraction II and Fraction m showed greater proliferative effect of mouse splenocytes stimulated by concanavalin A (Con A) than that of whole whey or Fraction L Proliferative effect of mouse splenocytes stimulated by phytohemagglutinin (PHA) was the highest when Fraction II was supplemented Proliferative effect of the colostral whey fractions on mouse splenocytes by stimulation of lipopolysaccharide (LPS) was markedly enhanced by supplementation of Fraction II and Fraction m compared with whole whey and Fraction L It was estimated that colostral whey fraction containing IGF-I positively affected proliferation of mouse splenocyte.

• Asian-Australasian Journal of Animal Sciences
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• 제19권2호
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• pp.297-304
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• 2006

Insulin-like growth factor-I (IGF-I) rich fraction, collected components between 1 kDa and 30 kDa, was fractionated from bovine colostral whey using an ultrafiltration membrane. IGF-I was confirmed in the collected IGF-I rich fraction by both SDS-PAGE and Western blotting. The concentration of IGF-I in the IGF-I rich fraction was 10 ng/mg protein. One hundred microliters of the reconstituted IGF-I rich fraction was intraperitoneally injected into ICR male mice for 2 weeks at 24 h intervals. The functions of peritoneal macrophages, including phagocytosis, interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$ production, and nitric oxide and hydrogen peroxide production, were enhanced significantly by the administration of the IGF-I rich fraction in a dose-dependent manner (p<0.01). The proliferation of Concanavalin (Con) A-stimulated and Lipopolysaccharide (LPS)-stimulated splenocytes was also determined to have been enhanced significantly by the administration of the IGF-I rich fraction in a dose-dependent manner (p<0.01). Our results indicate that the administration of IGF-I rich fraction obtained from bovine colostral whey enhances both innate and acquired immunity for ICR male mice.

• 한국식품과학회지
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• 제34권4호
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• pp.694-699
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• 2002

본 연구에서는 분만 후 5일 이내에 분비되는 젖소 초유의 whey 및 whey 분획이 Th1 cell의 증식에 어떤 영향을 미치는지, 그리고 Th1 cell 증식에 직접적으로 관여하는 whey 분획이 macrophage의 $TNF-{\alpha}$ 분비에 미치는 영향을 조사하였다. 초유 whey를 ultrafiltration으로 분자량별로 분획한 결과, 단백질 성분의 회수율은 Fr. I, II, III가 각각 72%, 17.7%, 10.2%였으며, 당 성분의 회수율은 Fr. I, II, III가 각각 22.1%, 7.4%, 70.1%였다. Fr. II를 재분획한 Fr. P의 단백질 회수율과 Fr. O의 당 회수율은 각각 86.9%, 88.8%였다. 각각의 whey 분획의 농도 대비 Th1 cell 증식 효과를 검증한 결과, 1 mg/mL 농도에서 Fr. II가 Th1 cell을 가장 많이 자극시켰으며, 세포증식율은 67.1%였으나, Fr. II의 단백질 및 올리고당 분획의 세포증식효과는 없는 것으로 나타났다. Whey의 각각의 분획을 이용하여 $TNF-{\alpha}$ 분비 능력을 조사한 결과, Fr. O가 양성대조구로 사용한 LPS보다 약 80% 이상의 $TNF-{\alpha}$분비 유도 능력이 있는 것으로 나타났다.

• 한국축산식품학회지
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• 제38권1호
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• pp.52-63
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• 2018

Cheese consumption has been gradually increased in China. However, both the manufacturing process of cheese and the utilization of its main by-product were not well developed. Based on the sensory evaluation, Box-Behnken Design (BBD) was performed in the present study to optimize the cheese processing, which was proved more suitable for Chinese. The optimal parameters were: rennet 0.052 g/L, start culture 0.025 g/L and $CaCl_2$ 0.1 g/L. The composition analysis of fresh bovine milk and whey showed that whey contained most of the soluble nutrients of milk, which indicated that whey was a potential resource of cyclic adenosine-3', 5'-monophosphate (cAMP). Thus, the cAMP was isolated from whey, the results of high-performance liquid chromatography (HPLC) analysis showed that the macroporous adsorption resins (MAR) D290 could increase the concentration of cAMP from $0.058{\mu}mol/mL$ to $0.095{\mu}mol/mL$. We firstly purified the cAMP from the whey, which could become a new source of cAMP.

• Food Science and Biotechnology
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• 제14권1호
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• pp.131-136
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• 2005

The selective extraction behavior of lactoferrin (Lf) from whey protein mixture was examined using reverse micelles formed by the cationic surfactant, cetyldimethylammonium bromide (CDAB). The major whey proteins, including ${\beta}$-lactoglobulin, ${\alpha}$-lactalbumin and bovine serum albumin, were solubilized from aqueous phase to organic phase while Lf was recovered in the aqueous phase. The solubilization behaviors of the proteins were manipulated by the process parameters such as the pH and salt concentration of the aqueous phase and the surfactant concentration in the organic phase. Efficient forward extraction was achieved with sodium borate buffer (50 mM, pH 9) containing 50 mM KCl and organic phase containing 100 mM CDAB. Based on SDS-PAGE and densitometry, about 96% of the initial Lf remained in the aqueous phase after forward extraction. The dialyzed Lf fully maintained its bacteriostatic activity against E. coli O157:H7.