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KMID : 1094720060110050442
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Biotechnology and Bioprocess Engineering
2006 Volume.11 No. 5 p.442 ~ p.448
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High-yield production of functional human lactoferrin in transgenic cell cultures of siberian ginseng ( Acanthopanax senticosus )
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Jo Seung-Hyun
Kwon Suk-Yoon
Park Doo-Sang
Yang Kyoung-Sil
Kim Jae-Whune
Lee Ki-Teak
Kwak Sang-Soo
Lee Haeng-Soon
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Abstract
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Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducibleSWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.
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KEYWORD
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cell culture, high expression promoter, human lactoferrin, medicinal plant, recombinant protein
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