Biodegradable poly-3-hydroxybutyrate as a shielding carrier for a plant-protecting MF3 protein from Pseudomonas fluorescens

Popletaeva, S.B.; Voinova, T.M.; Kartashov, M.I.; Pasechnik, T.D.; Arslanova, L.R.; Shcherbakova, L.A.; Statsyuk, N.V.; Bonartsev, A.P.; Zernov, A.L.; Bonartseva, G.A.; Dzhavakhiya, V.G.

Авторы: Popletaeva S.B., Voinova T.M., Kartashov M.I., Pasechnik T.D., Arslanova L.R., Shcherbakova L.A., Statsyuk N.V., Bonartsev A.P., Zernov A.L., Bonartseva G.A., Dzhavakhiya V.G.
Журнал: Biointerface Research in Applied Chemistry
Том: 8
Номер: 3
Год издания: 2018
Первая страница: 3224
Последняя страница: 3231
Аннотация: Boosting of plant immunity to improve its resistance to pathogens is a promising approach to increase crop capacity and reduce pesticide press on the environment. A typical scenario of such approach is based on the use of elicitors able to activate key plant defense mechanisms. MF3, a highly thermostable FKBP-type peptidyl prolyl cis/trans isomerase from Pseudomonas fluorescens, has a proved eliciting activity regarding various crops. However, for field application, this enzyme should be protected against degrading action of sunlight, as well as plant and microbial proteinases. In this study we examined a possibility to encapsulate MF3 using biodegradable poly-3-hydroxybutyrate (PHB) and evaluated the efficacy of the resulted MF3 shielding against UV radiation and enzymatic proteolysis, as well as the resistance-eliciting activity of the encapsulated protein. PHB-based microparticles loaded with MF3 in a complex with low-molecular chitosan and 500-kDa dextran were obtained using a Solid/Oil/Water technique followed with a spray drying and freeze frying. The resulting microparticles consisted of PHB (80%), chitosan (4%), dextran (6%), and MF3 (10%) and had a size of 10-25 μm. The effective loading capacity was ~10%; in vitro kinetic study showed that 89% of the total loaded protein was released within the first 24 h, while the rest of the protein slowly released within next 12 days. A comparison of the resistance-eliciting activity of free and encapsulated MF3 was carried out using two model “plant-pathogen” systems, tobacco/tobacco mosaic virus (TMV) and tobacco/Alternaria longipes. In both models, activity of encapsulated MF3 was similar to that of free MF3 of the same concentration and resulted in the reduction of infection level by 30.3–36.8% for A. longipes and 66.8–68.4% for TMV. In vitro assessment of the effect of UV irradiation and proteinase K treatment on the activity of free and encapsulated MF3 showed a clear shielding effect of PHB. Even after 8 h of UV treatment, when free MF3 completely lost its resistance-eliciting activity, encapsulated MF3 still provided almost 50% suppression of TMV infection. Treatment with proteinase K (20 and 100 μg/mL) resulted in significant loss of free MF3 protecting efficiency (22.9 and 35.5%, respectively), while for encapsulated MF3 this loss made only 2.6 and 12.2%, respectively. These results confirmed the encapsulation procedure did not impair the protective activity of MF3, while provided a successful release of this protein from the complex and its delivery to putative receptors of plant cells. The study has shown a principal possibility to produce PHB-based MF3 preparations characterized by improved resistance of their bioactive component to adverse biotic and abiotic factors and possessed a significant protective activity in relation to the tested plant pathogens. The further improvement of the encapsulation technology and the characterization of encapsulated MF3 using other model systems and whole plants are planned.
Добавил в систему: Бонарцев Антон Павлович

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Biodegradable poly-3-hydroxybutyrate as a shielding carrier for a plant-protecting MF3 protein from Pseudomonas fluorescensстатья Исследовательская статья

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