کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6491043 | 43390 | 2015 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Novel enzymatic synthesis of spacer-linked Pk trisaccharide targeting for neutralization of Shiga toxin
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موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
A novel alkyl spacer-conjugated derivative of Pk trisaccharide (Pk), one of the active receptors of Shiga toxins (Stxs; Stx1 and Stx2) produced by pathogenic Escherichia coli (STEC), was designed and synthesized by a combination of cellulase-mediated condensation from Trichoderma reesei and α1,4-galactosyltransferase (LgtC) from Neisseria gonorrhoeae. The specific activity of N. gonorrhoeae LgtC was 66 U/mg, which was 13-fold higher than that from N. meningitidis expressed in E. coli. 5-trifluoroacetamidopentyl-β-Pk (TFAP-Pk) was synthesized (yield of 86%, based on the amount of TFAP-lactose added) and its binding to Stx1a-B and Stx2a-B was evaluated. The dissociation constants (KDs) of Stx1a-B and Stx2a-B to the spacer-linked Pk, immobilized on a CM5 sensor chip, were 6.8 Ã 10â6 M (kon = 4.1 Ã 101 Mâ1 Sâ1, koff = 2.8 Ã 10â4 Sâ1) and 2.2 Ã 10â5 M (kon = 3.9 Ã 102 Mâ1 Sâ1, koff = 8.6 Ã 10â3 Sâ1), respectively. This result suggests that the monovalent Pk-derivative, conjugated to a pentylamino group, represents a promising Stx-neutralizing agent. This cellulase-mediated condensation using cellulase and glycosyltransferase is a valuable tool for the synthesis of spacer-linked oligosaccharide.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 209, 10 September 2015, Pages 50-57
Journal: Journal of Biotechnology - Volume 209, 10 September 2015, Pages 50-57
نویسندگان
Tatsuya Kato, Takahiro Oizumi, Makoto Ogata, Akiko Murakawa, Taichi Usui, Enoch Y. Park,