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Sci Rep ; 11(1): 17966, 2021 09 09.
Artículo en Inglés | MEDLINE | ID: mdl-34504153

RESUMEN

Hyaluronic acid (HA), a unique polysaccharide with excellent Physico-chemical properties, is broadly used in pharmaceutical, biomedical, and cosmetic fields. It is widely present in all vertebrates, certain bacterial strains, and even viruses while it is not found in plants, fungi, and insects. HA is naturally synthesized by a class of integral membrane proteins called Hyaluronic acid synthase (HAS). Thus far, industrial production of HA is carried out based on either extraction from animal sources or large-scale microbial fermentation. The major drawbacks to using these systems are contamination with pathogens and microbial toxins. Recently, the production of HA through recombinant systems has received considerable attention. Plants are eco-friendly ideal expression systems for biopharmaceuticals production. In this study, the optimized human hyaluronic acid synthase2 (hHAS2) sequence was transformed into Nicotiana tabacum using Agrobacterium rhizogenes. The highest rhHAS2 concentration of 65.72 ng/kg (wet weight) in transgenic tobacco hairy roots was measured by the human HAS2 ELISA kit. The HA production in the transgenic hairy roots was verified by scanning electron microscope (SEM) and quantified by the HA ELISA kit. The DPPH radical scavenging activity of HA with the highest concentration of 0.56 g/kg (wet weight) showed a maximum activity of 46%. Gel Permeation Chromatography (GPC) analyses revealed the high molecular weight HA (HMW-HA) with about > 0.8 MDa.


Asunto(s)
Productos Biológicos/metabolismo , Hialuronano Sintasas/metabolismo , Ácido Hialurónico/biosíntesis , Nicotiana/genética , Nicotiana/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Agrobacterium/genética , Secuencia de Bases , Productos Biológicos/química , Cromatografía en Gel/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Hialuronano Sintasas/genética , Ácido Hialurónico/química , Microscopía Electrónica de Rastreo/métodos , Peso Molecular , Plantas Modificadas Genéticamente , Transformación Genética
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