• Journal of Veterinary Clinics
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• v.24 no.3
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• pp.305-307
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• 2007

Lactoferrin-binding proteins (LBP) has not been well characterized in Streptococcus uberis isolated from milk of bovine mastitis and to date this protein is considered to be an important virulence factor in Streptococcal mastitis. To determine the more efficient extraction method of LBP from four S. uberis strains, we used two different extraction methods (mutanolysin and sodium dodecyl sulfate) in this study. Bacterial proteins extracted were electrophoresed by 10% polyacrylamide gels in the presence of sodium deodecyl sulfate and gels were transferred onto nitrocellulose membrane. Rabbit anti-bovine lactoferrin antibody and HRP-conjugated donkey anti-rabbit IgG antibody were used to detect LBP. This Western blotting analysis demonstrates that extraction method with SDS extracted 110 kDa and 112 kDa LBPs more efficiently compared to the mutanolysin extraction method.

• Food Science of Animal Resources
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• v.25 no.2
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• pp.232-237
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• 2005

Lactoferrin is a member of the transferrin family of iron-binding glycoproteins. It is originally found in milk. In addition to its antibacterial and antiviral activities, lactoferrin has many other biological functions include anti-inflammatory properties, antitumor, cell growth-promoting activity as well as antioxidant effect In the present study, we report the production of recombinant bovine lactoferrin and lactoferrin N-lobe in the Rhodococcus erythropolis (R erythropolis) using pTip vector. The expression level was investigated in various range of temperature, and we could successfully expressed the bovine lactoferrin and lactoferrin N-lobe in R erythropolis at low temperature. The recombinant proteins were purified by Nickel-Nitrolotriacetic acid (Ni-NTA). The purified proteins were confirmed by SDS-PAGE and Western blot, which indicating that the recombinant proteins have a molecular weight of 80kDa and 43kDa for bovine lactoferrin and lactoferrin N-lobe, respectively.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.271-279
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• 2015

Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in physiological secretions of mammals. LF shows antibacterial, antiviral and antifungal activities. In the present study, a gene encoding the N-terminal lobe of human lactoferrin (hLF) was isolated, cloned and expressed in methylotrophic yeast, Pichia pastoris. The recombinant hLF-N (rhLF-N) protein was secreted into the culture medium at the level of $458{\mu}g/ml$ in 3 L fermentor. The size of purified hLF-N was estimated as 35 kDa when analyzed by SDS-PAGE and western blotting. The rhLF-N was further confirmed by immunodiffusion using the anti-hLF polyclonal antibody. The expression profile analysis by qRT-PCR showed that the relative mRNA expression of rhLF-N was maximal after 2-3 days of methanol induction and reduced gradually at 4 days. The purified rhLF-N showed broad antibacterial activities against the pathogens such as Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, and Salmonella typhimurium. However, rhLF-N showed relatively lower activity when compared to peptides derived from LF. In spite of this weak activity, the rhLF-N expressed in P. pastoris might be more advantageous for the industrial application, because rhLF-N is secreted into the culture medium and the production can also be increased by optimization of culture conditions.

• Journal of Plant Biotechnology
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• v.32 no.3
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• pp.209-215
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• 2005

Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.10
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• pp.551-558
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• 2017

To study and validate tissue-specific promoters and vectors, it is important to develop cell culture systems that retain the tissue and species specificity. Such systems are attractive alternatives to transgenic animal models. This study established a line of porcine mammary gland epithelial cells (PMECs) from a primary culture based on the cellular morphology and mRNA levels of porcine beta-casein (CSN2). The selected PMECs were stained with the cytokeratin antibody, and were shown to express milk protein genes (CSN2, lactoferrin, and whey acidic protein). In addition, to confirm the acini structure of PMEC932-7 in 3D culture, live cells were stained with SYTO-13 dye, which binds to nucleic acid. The acini of these PMECs on matrigel were formed by the aggregation of peripheral cells and featured a hollow lumens. The system was demonstrated by testing the effects of the culture conditions to cell culture including cell density and matrigel methods of the PMECs. These results suggest that PMECs possess the genetic and structural features of mammary epithelial cells.

• Journal of Dairy Science and Biotechnology
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• v.16 no.2
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• pp.106-118
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• 1998

Whey is a natural product obtained during cheese production. With the advent of new technology, whey protein concentrates and whey fractions have become readily available and versatile food ingredients. Whey protein concentrates are highly functional ingredients that have gelling, emulsifying, whipping, water-binding and fat-replacement properties. New fractions derived from whey (such as alpha-lactalbumin, lactoferrin, lactoperoxidase and peptides) attract considerable interest worldwide because of their bioactive or health-enhancing properties. Some of these fractions also find new uses as natural antibiotic, natural preservative and immunity-enhancing agents. With the growth of the functional foods industry sector, an increasing number of manufacturers take advantage of whey's nutritional and functional benefits to develop successful new products. The United States is the world's largest single producer and exporter of whey products. In 1997, more than 1 million metric tons of whey products were manufacturers in the U.S.