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Enhancement of activity and thermostability of Aspergillus niger ATCC 10864 phytase A through rational designстатья
- Авторы: Dotsenko Anna, Rozhkova Aleksandra, Zorov Ivan, Korotkova Olga, Sinitsyn Arkady
- Журнал: Biochemical and Biophysical Research Communications
- Том: 634
- Год издания: 2022
- Издательство: Academic Press
- Местоположение издательства: United States
- Первая страница: 55
- Последняя страница: 61
- DOI: 10.1016/j.bbrc.2022.10.010
- Аннотация: AbstractAspergillus niger ATCC 10864 phytase A was produced in Penicillium verruculosum. The enzyme was found to have two pH optima of 2.5 and 5.0, as well as a T-optimum of 50–55 °C. Two amino acid substitutions, A76M and S265P, were designed for improvement in thermostability, and two more, N300K and D363N, were designed for improvement in enzyme activity. The most thermostable variant, S265P, was characterized by a 3.8-fold increase in time of half-life at 55 °C and a 1.2-fold increase in residual activity at 90 °C compared to the wild-type. The most active variant, D363N, was 1.7-times more active at 40 °C and retained 1.3-times higher residual activity at 90 °C compared to the wild-type. The obtained results revealed the importance of substitutions with proline in α-helixes for the thermostability improvement of phytases. Also, the importance of sequence motif 361HDN363 was demonstrated with relevance to values of catalytic parameters.Highlights• Activity at pH 2.0–2.5 and 5.5 was improved through design of active center.• Improvement in activity was 1.3-fold at pH 2.5 and 1.7-fold at pH 5.5.• Thermostability was improved through filling of intramolecular cavities.• Tm was improved by 0.9–2.0 °C, thermostability at 90 °C was improved by 1.2-fold.
- Добавил в систему: Синицын Аркадий Пантелеймонович
Прикрепленные файлы
| № | Имя | Описание | Имя файла | Размер | Добавлен |
|---|---|---|---|---|---|
| 1. | Первая страница | Article_Dotsenko_Enhancement_astability_of_phytase_BBRC_202… | 169,2 КБ | 14 декабря 2022 [ZorovIN] | |
| 2. | Article_Dotsenko_Enhancement_activity_and_thermostability_o… | Article_Dotsenko_Enhancement_activity_and_thermostability_o… | 822,7 КБ | 27 октября 2022 [aps306cellulas] |