High-level production of lactostatin, a hypocholesterolemic peptide, in transgenic rice using soybean A1aB1b as carrier
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Hypercholesterolemia, a form of cardiovascular disease, is one of the leading causes of deaths worldwide. Lactostatin (Ile-Ile-Ala-Glu-Lys), derived from β-lactoglobulin in cow’s milk, is a bioactive peptide with hypocholesterolemic activity higher than sitosterol, a known anti-hypercholesterolemic drug. Here, we successfully developed a transgenic rice accumulating a much higher level of lactostatin by inserting 29 IIAEK sequences into the structurally flexible (nonconserved) regions of soybean seed storage protein, A1aB1b, and introducing it into LGC-1 (low glutelin content mutant 1) as host variety. A1aB1b containing 29 lactostatins was expressed in the endosperm of rice seed cells by using seed specific promoters and sorted into novel compartments distinct from normal PB-I (ER-derived protein body) and PB-II (protein storage vacuoles). Transgenic rice seeds accumulated approximately 2 mg of lactostatins/g of dry seeds, which is relatively high compared with previous reports. Our findings suggest that the introduction of a high copy number of bioactive peptide into seed storage proteins as carrier is one of the effective means in producing higher amounts of bioactive peptides in rice.
KeywordsBioactive peptide Rice Seed storage protein Lactostatin
We thank DASH (Development and Assessment of Sustainable Humanosphere) system and Prof. Kazufumi Yazaki of Kyoto University for supporting the development of the transgenic crop. We also thank Prof. Reiko Urade (Kyoto University) for the warm encouragement and Ms. M. Sawada and Ms E. Okuda for technical assistance. This research was supported by the research grant, “Genomics and Agricultural Innovation, GMC0008” from the Ministry of Agriculture, Forestry and Fisheries of Japan to M. Kuroda, S. Nagaoka, F. Takaiwa, S. Utsumi and N. Maruyama, and the Asahi Glass Foundation to N Maruyama and S Utsumi.
- Bridget BK, Fuster V (2010) Promoting cardiovascular health in the developing world: a critical challenge to achieve global health. National Academies Press, Washington, DCGoogle Scholar
- Chobanian AV, Bakris GL, Black HR, Cushman WC, Green LA, Izzo JL Jr, National Heart, Lung and Blood Institute Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure, National High Blood Pressure Education Program Coordinating Committee et al (2003) The seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure: the JNC 7 report. J Am Med Assoc 289:2560–2572CrossRefGoogle Scholar
- Gaziano TA, Bitton A, Anand S, Abrahams-Gessel S, Murphy A (2010) Growing epidemic of coronary heart disease in low- and middle-income countries. Curr Probl Cardiol 35:72–115Google Scholar
- Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
- Osborne TB (1907) The proteins of the wheat kernel. Carnegie Institution of Washington, Publ No. 84, 119 ppGoogle Scholar
- Saito H, Shigemitsu T, Yamasaki R, Sasou A, Goto F, Kishida K, Kuroda M, Tanaka K, Morita S, Satoh S, Masumura T (2012) Formation mechanism of the internal structure of type I protein bodies in rice endosperm: relationship between the localization of prolamin species and the expression of individual genes. Plant J 70:1043–1055PubMedCrossRefGoogle Scholar
- Takaiwa F (2005) Health-promoting transgenic rice suppressing life-related disease and type-I allergy. In: Toriyama K, Heong KL, Hardy B (eds) Rice is life: scientific perspectives for the 21st century, proceedings of the world rice research conference, Tsukuba, Japan, pp 102–105Google Scholar