JJPHYSIOL : Vol. 53 (2003) , No. 1 65-69

Development of a High-Level Expression System for Deuterium-Labeled Human Serum Albumin

Mihoko Tomida¹⁾, Masashi Kimura, Kazuo Kuwata, Tomoya Hayashi, Yukio Okano and Seiichi Era

1) Department of Biochemistry, Gifu University School of Medicine

Abstract: We have developed an expression system for recombinant human serum albumin (rHSA) using methylotrophic yeast Pichia pastoris Mut⁺ transformants together with the multiple cross-over integration of the vector containing human serum albumin (HSA). After 86 h of methanol induction, the secreted rHSA reached levels of ∼320 mg/l in 100% H₂O medium and ∼180 mg/l in 70% D₂O/30% H₂O (v/v) medium in a fed-batch fermenter. The structures of the obtained rHSA and plasma-derived HSA (pHSA) were virtually identical as viewed from various physicochemical techniques such as HPLC, SDS gel electrophoresis, and CD. NMR peaks of the partially deuterium (D)-labeled rHSA (DrHSA) were quite sharp compared to those of pHSA due to suppression of the intramolecular nuclear Overhauser effect, promising further structural studies of the whole HSA molecule in the solution state using the recent NMR techniques.

Key words: human serum albumin, partial D-labeling, Pichia pastoris

	To cite this article:
	Mihoko Tomida, Masashi Kimura, Kazuo Kuwata, Tomoya Hayashi, Yukio Okano and Seiichi Era: “Development of a High-Level Expression System for Deuterium-Labeled Human Serum Albumin”, JJP Vol. 53: 65-69, (2003) .

	doi:10.2170/jjphysiol.53.65
	JOI JST.JSTAGE/jjphysiol/53.65