Expression analysis of human beta-secretase in transgenic tomato fruits = 형질전환 감자에서 베타시크리테아제의 발현분석

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Title
Expression analysis of human beta-secretase in transgenic tomato fruits = 형질전환 감자에서 베타시크리테아제의 발현분석
Author(s)
Hyun Soon Kim; J W Youm; Ki Beom Moon; Jang Ho Ha; Y H Kim; Hyouk Joung; Jae Heung Jeon
Bibliographic Citation
Protein Expression and Purification, vol. 82, no. 1, pp. 125-131
Publication Year
2012
Abstract
An emerging strategy in biomanufacturing involves using transgenic plants to express recombinant pharmaceutical and industrial proteins in large quantities. β-Site APP cleaving enzyme 1 (β-secretase 1, BACE1) is an enzyme involved in the abnormal production of Aβ42, the major component of senile plaques in Alzheimer's disease (AD). Thus, BACE1 represents a key target protein in the development of new potential drugs to treat Alzheimer's disease. We aimed to develop a tomato-derived recombinant BACE1 (rBACE1) protein to serve as a vaccine antigen that would promote an immune response. We utilized a plant expression cassette, pE8BACE, to optimize BACE1 expression in tomato fruits. Polyemerase chain reaction and Southern blot analyses verified integration of the BACE1 gene into the plant genome. Northern and Western blot analyses demonstrated successful mRNA and protein expression of rBACE1, respectively; the Sensizyme assay kit estimated the expression level of rBACE1 protein at 136 ± 7 ng mg -1 total soluble protein. The tomato-derived rBACE1 retains its activity for a long storage period at cool or room temperature, and is highly resistant to degradation in conditions such as low acidity. Tomato-derived rBACE1 was severely degraded by heat or boiling. The proteolytic activity of tomato-derived rBACE1, confirmed by fluorescence resonance transfer assay, was similar to that of a commercial sample of Escherichia coli-derived BACE1.
Keyword
β-SecretaseAlzheimer's vaccineLycopersicon esculentumRecombinant proteinTransgenic plant
ISSN
1046-5928
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.pep.2011.11.012
Type
Article
Appears in Collections:
Division of Research on National Challenges > Plant Systems Engineering Research > 1. Journal Articles
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