期刊
JOURNAL OF BIOTECHNOLOGY
卷 270, 期 -, 页码 39-43出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2018.01.011
关键词
Biocatalysis; Glyoxyl agarose; Stabilization; Floxuridine; Molecular cloning
资金
- National Agency of Scientific and Technological Promotion [PICT 2013-2658, PICT 2014-3438]
- National Scientific and Technical Research Council [PIP 2014-KA5-00805]
- National University of Quilmes [PUNQ 1409/15]
- IKERBASQUE Foundation
- COST Action Systems Biocatalysis [CM1303]
The 2'-N-deoxyribosyltransferases [NDT; EC 2.4.2.6] are a group of enzymes widely used as biocatalysts for nucleoside biosynthesis. In this work, the molecular cloning, expression and purification of a novel NDT from Lactobacillus animalis (LaNDT) have been reported. On the other hand, biocatalyst stability has been significantly enhanced by multipoint covalent immobilization using a hetero-functional support activated with nickel-chelates and glyoxyl groups. The immobilized enzyme could be reused for more than 300 h and stored during almost 3 months without activity loss. Besides, the obtained derivative (Ni2+-Gx-LaNDT) was able to biosynthesize 88 mg floxuridine/g biocatalyst after 1 h of reaction. In this work, a green bioprocess by employing an environmentally friendly methodology was developed, which allowed the obtaining of a compound with proven anti-tumor activity. Therefore, the obtained enzymatic biocatalyst meets the requirements of high activity, stability, and short reaction times needed for low-cost production in a future preparative application.
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