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1.
Mol Plant ; 14(7): 1149-1167, 2021 07 05.
Artículo en Inglés | MEDLINE | ID: mdl-33857689

RESUMEN

The proteolytic degradation of the photodamaged D1 core subunit during the photosystem II (PSII) repair cycle is well understood, but chlorophyll turnover during D1 degradation remains unclear. Here, we report that Arabidopsis thaliana CHLOROPHYLLASE 1 (CLH1) plays important roles in the PSII repair process. The abundance of CLH1 and CLH2 peaks in young leaves and is induced by high-light exposure. Seedlings of clh1 single and clh1-1/2-2 double mutants display increased photoinhibition after long-term high-light exposure, whereas seedlings overexpressing CLH1 have enhanced light tolerance compared with the wild type. CLH1 is localized in the developing chloroplasts of young leaves and associates with the PSII-dismantling complexes RCC1 and RC47, with a preference for the latter upon exposure to high light. Furthermore, degradation of damaged D1 protein is retarded in young clh1-1/2-2 leaves after 18-h high-light exposure but is rescued by the addition of recombinant CLH1 in vitro. Moreover, overexpression of CLH1 in a variegated mutant (var2-2) that lacks thylakoid protease FtsH2, with which CLH1 interacts, suppresses the variegation and restores D1 degradation. A var2-2 clh1-1/2-2 triple mutant shows more severe variegation and seedling death. Taken together, these results establish CLH1 as a long-sought chlorophyll dephytylation enzyme that is involved in PSII repair and functions in long-term adaptation of young leaves to high-light exposure by facilitating FtsH-mediated D1 degradation.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Hidrolasas de Éster Carboxílico/metabolismo , Proteínas de Choque Térmico/metabolismo , Luz , Metaloendopeptidasas/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Hojas de la Planta/efectos de la radiación , Arabidopsis/crecimiento & desarrollo , Arabidopsis/efectos de la radiación , Fotosíntesis , Hojas de la Planta/enzimología , Protectores contra Radiación , Tilacoides/metabolismo
2.
Mol Biotechnol ; 60(7): 492-505, 2018 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-29796788

RESUMEN

Natural production of anti-cancer drug taxol from Taxus has proved to be environmentally unsustainable and economically unfeasible. Currently, bioengineering the biosynthetic pathway of taxol is an attractive alternative production approach. 10-deacetylbaccatin III-10-O-acetyl transferase (DBAT) was previously characterized as an acyltransferase, using 10-deacetylbaccatin III (10-DAB) and acetyl CoA as natural substrates, to form baccatin III in the taxol biosynthesis. Here, we report that other than the natural acetyl CoA (Ac-CoA) substrate, DBAT can also utilize vinyl acetate (VA), which is commercially available at very low cost, acylate quickly and irreversibly, as acetyl donor in the acyl transfer reaction to produce baccatin III. Furthermore, mutants were prepared via a semi-rational design in this work. A double mutant, I43S/D390R was constructed to combine the positive effects of the different single mutations on catalytic activity, and its catalytic efficiency towards 10-DAB and VA was successfully improved by 3.30-fold, compared to that of wild-type DBAT, while 2.99-fold higher than the catalytic efficiency of WT DBAT towards 10-DAB and Ac-CoA. These findings can provide a promising economically and environmentally friendly method for exploring novel acyl donors to engineer natural product pathways.


Asunto(s)
Acetiltransferasas/genética , Alcaloides/biosíntesis , Antineoplásicos Fitogénicos/biosíntesis , Taxus/enzimología , Acetiltransferasas/química , Acetiltransferasas/metabolismo , Alcaloides/economía , Antineoplásicos Fitogénicos/economía , Bioingeniería , Vías Biosintéticas , Biología Computacional , Análisis Costo-Beneficio , Ingeniería Genética , Modelos Moleculares , Mutagénesis , Paclitaxel/biosíntesis , Paclitaxel/economía , Especificidad por Sustrato , Taxoides/economía , Taxoides/metabolismo , Taxus/química , Taxus/genética , Taxus/metabolismo , Compuestos de Vinilo/química , Compuestos de Vinilo/metabolismo
3.
Artículo en Chino | MEDLINE | ID: mdl-24830207

RESUMEN

When the digital microscopy interactive system is applied to parasitology experiment teaching, students can submit their experiment reports in two ways: a paper document on a drawing paper or an electronic document taken images with the system. Submission of a paper report needs more time but requires the students to work more carefully, and an electronic document allows them to have more time to observe the specimen and work in a higher efficiency. It would be better to ask students to do both.


Asunto(s)
Parasitología/educación , Microscopía
4.
Artículo en Chino | MEDLINE | ID: mdl-21351560

RESUMEN

The active ingredient of Huyinling, a combination of Chinese traditional medicine, was extracted by five different ethanol concentrations (40%-80%). There were seven groups named as five Huyinling ethanol extract groups (40%, 50%, 60%, 70%, and 80%), metronidazole group and blank control. Each Huyinling ethanol extract group was further divided into five subgroups with final concentration of 6.25, 12.5, 25, 50, and 100 mg/ml, respectively. Metronidazole group was given 10 microg/ml of the drug. Each group had 4 wells with 125 microl T. tenax(2 x 10(5)/ml). At 12 h, 24 h and 48 h after drug treatment, the anti-T. tenax effect of Huyinling ethanol extract was tested by microscope counting method. At 24 h the effect of Huyinling on T. tenax was examined with methyl thiazolyl tetrazolium (MTl) assay. The results showed that the higher concentration of Huyinling ethanol extract, the better effect on anti-T. tenax. 60% Huyinling ethanol extract group with concentrations of 6.25 mg/ml and 12.5 mg/ml showed higher anti-T. tenax effect than other groups (P < 0.01). The ethanol extract of Huyinling granules has a remarkable effect on T. tenax, and among the groups, 60% ethanol extract shows the best anti-T. tenax activity.


Asunto(s)
Antitricomonas/farmacología , Medicamentos Herbarios Chinos/farmacología , Trichomonas/efectos de los fármacos
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