Expression, Purification, and Enzyme Activity Assay of Phytoene Synthase In Vitro

2019 ◽  
pp. 39-52
Author(s):  
Maurizio Camagna ◽  
Ralf Welsch
Keyword(s):  
Enzyme Activity ◽  
Phytoene Synthase ◽  
Activity Assay ◽  
Enzyme Activity Assay

Expression, Purification and in vitro Enzyme Activity Assay of Plant Derived GTPase

BIO-PROTOCOL ◽  
2015 ◽  
Vol 5 (22) ◽  
Author(s):  
Annemarie Gl�ckner ◽  
Christian Voigt
Keyword(s):  
Enzyme Activity ◽  
Activity Assay ◽  
Enzyme Activity Assay

scholarly journals The Alternative Route to Heme in the Methanogenic ArchaeonMethanosarcina barkeri

Archaea ◽  
2014 ◽  
Vol 2014 ◽  
pp. 1-13 ◽  
Author(s):  
Melanie Kühner ◽  
Kristin Haufschildt ◽  
Alexander Neumann ◽  
Sonja Storbeck ◽  
Judith Streif ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Feedback Regulation ◽  
Methanosarcina Barkeri ◽  
Heme Biosynthesis ◽  
Alternative Route ◽  
Activity Assay ◽  
Enzyme Activity Assay ◽  
Living Organisms

In living organisms heme is formed from the common precursor uroporphyrinogen III by either one of two substantially different pathways. In contrast to eukaryotes and most bacteria which employ the so-called “classical” heme biosynthesis pathway, the archaea use an alternative route. In this pathway, heme is formed from uroporphyrinogen III via the intermediates precorrin-2, sirohydrochlorin, siroheme, 12,18-didecarboxysiroheme, and iron-coproporphyrin III. In this study the heme biosynthesis proteins AhbAB, AhbC, and AhbD fromMethanosarcina barkeriwere functionally characterized. Using anin vivoenzyme activity assay it was shown that AhbA and AhbB (Mbar_A1459 and Mbar_A1460) together catalyze the conversion of siroheme into 12,18-didecarboxysiroheme. The two proteins form a heterodimeric complex which might be subject to feedback regulation by the pathway end-product heme. Further, AhbC (Mbar_A1793) was shown to catalyze the formation of iron-coproporphyrin IIIin vivo. Finally, recombinant AhbD (Mbar_A1458) was produced inE. coliand purified indicating that this protein most likely contains two [4Fe-4S] clusters. Using anin vitroenzyme activity assay it was demonstrated that AhbD catalyzes the conversion of iron-coproporphyrin III into heme.

Expression and Enzyme Activity Assay of pfu with Molecular Chaperone

2010 ◽  
Vol 31 (6) ◽  
pp. 499-503
Author(s):  
Hai-Jun ZHANG ◽  
Jun YANG ◽  
Xiao-Guang LIU ◽  
Xiang-Yang HU
Keyword(s):  
Enzyme Activity ◽  
Molecular Chaperone ◽  
Activity Assay ◽  
Enzyme Activity Assay

Impact of enzyme activity assay on indication in liver transplantation for ornithine transcarbamylase deficiency

2012 ◽  
Vol 105 (3) ◽  
pp. 404-407 ◽  
Author(s):  
Taiichi Wakiya ◽  
Yukihiro Sanada ◽  
Taizen Urahashi ◽  
Yoshiyuki Ihara ◽  
Naoya Yamada ◽  
...  
Keyword(s):  
Liver Transplantation ◽  
Enzyme Activity ◽  
Ornithine Transcarbamylase ◽  
Ornithine Transcarbamylase Deficiency ◽  
Activity Assay ◽  
Enzyme Activity Assay

Resistance in Smallflower Umbrella Sedge (Cyperus difformis L.) to Acetolactate Synthase-Inhibiting Herbicides in Rice - First Case in India

2021 ◽  
pp. 1-22
Author(s):  
Vijay K. Choudhary ◽  
Seshadri S. Reddy ◽  
Subhash K. Mishra ◽  
Bhumesh Kumar ◽  
Yogita Gharde ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Acetolactate Synthase ◽  
Activity Assay ◽  
Enzyme Activity Assay ◽  
Synthetic Auxin ◽  
Cyperus Difformis ◽  
First Case ◽  
Whole Plant ◽  
Plant Bioassay ◽  
Direct Seeded

Abstract Smallflower umbrella sedge is one of the problematic weeds in direct-seeded rice in India. Bispyribac-sodium (acetolactate synthase-inhibiting herbicide) is a commonly used in rice, but recently growers have reported lack of smallflower umbrella sedge control with this herbicide. An extensive survey was carried out in two rice growing states, Chhattisgarh and Kerala, where 53 putative bispyribac-sodium resistant (BR) biotypes were collected. Studies were conducted to confirm resistance to bispyribac-sodium and to test the efficacy of newly developed synthetic auxin herbicide florpyrauxifen-benzyl on putative BR biotypes. Whole-plant bioassay revealed that bispyribac-sodium is no longer effective. Of 53 putative BR biotypes, 17 biotypes survived recommended label rate of 25 g ai ha−1. Effective bispyribac-sodium rate required to control 50% of the plants in most of the BR biotypes (ED50) ranged from 19 to 96 g ha−1 whereas it was 10 g ha−1 in susceptible biotype. In two highly resistant biotypes, ED50 was beyond the maximum tested rate, 200 g ha−1. This suggests 2 to >20-fold resistance in BR biotypes. Acetolactate synthase (ALS) enzyme activity assay suggests altered target site as mechanism of resistance to bispyribac-sodium. This study confirms the first case of evolved resistance in smallflower umbrella sedge for bispyribac-sodium in India. However, the newly developed synthetic auxin, florpyrauxifen-benzyl effectively controlled all BR biotypes at the field use rate 31.25 g ae ha−1.

scholarly journals A High‐Throughput Mass Spectrometric Enzyme Activity Assay Enabling the Discovery of Cytochrome P450 Biocatalysts

2019 ◽  
Vol 58 (30) ◽  
pp. 10114-10119 ◽  
Author(s):  
Tristan de Rond ◽  
Jian Gao ◽  
Amin Zargar ◽  
Markus de Raad ◽  
Jack Cunha ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Cytochrome P450 ◽  
High Throughput ◽  
Mass Spectrometric ◽  
Activity Assay ◽  
Enzyme Activity Assay

Adenine/Au complex-dependent versatile electrochemical platform for ultrasensitive DNA-related enzyme activity assay

2018 ◽  
Vol 273 ◽  
pp. 760-770 ◽  
Author(s):  
Shaohua Ma ◽  
Yufang Hu ◽  
Qingqing Zhang ◽  
Zhiyong Guo ◽  
Sui Wang ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Activity Assay ◽  
Enzyme Activity Assay ◽  
Related Enzyme

scholarly journals Design and Synthesis of N-phenyl Phthalimides as Potent Protoporphyrinogen Oxidase Inhibitors

Molecules ◽  
2019 ◽  
Vol 24 (23) ◽  
pp. 4363
Author(s):  
Wei Gao ◽  
Xiaotian Li ◽  
Da Ren ◽  
Susu Sun ◽  
Jingqian Huo ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Enzyme Activity ◽  
Molecular Docking ◽  
Molecular Design ◽  
Design Strategy ◽  
Docking Studies ◽  
Activity Assay ◽  
Enzyme Activity Assay ◽  
Protoporphyrinogen Oxidase ◽  
Design And Synthesis

Protoporphyrinogen oxidase (PPO) has been identified as one of the most promising targets for herbicide discovery. A series of novel phthalimide derivatives were designed by molecular docking studies targeting the crystal structure of mitochondrial PPO from tobacco (mtPPO, PDB: 1SEZ) by using Flumioxazin as a lead, after which the derivatives were synthesized and characterized, and their herbicidal activities were subsequently evaluated. The herbicidal bioassay results showed that compounds such as 3a (2-(4-bromo-2,6-difluorophenyl) isoindoline-1,3-dione), 3d (methyl 2-(4-chloro-1,3-dioxoisoindolin-2-yl)-5-fluorobenzoate), 3g (4-chloro-2-(5-methylisoxazol-3-yl) isoindoline-1,3-dione), 3j (4-chloro-2-(thiophen-2-ylmethyl) isoindoline-1,3-dione) and 3r (2-(4-bromo-2,6-difluorophenyl)-4-fluoroisoindoline-1,3-dione) had good herbicidal activities; among them, 3a showed excellent herbicidal efficacy against A. retroflexus and B. campestris via the small cup method and via pre-emergence and post-emergence spray treatments. The efficacy was comparable to that of the commercial herbicides Flumioxazin, Atrazine, and Chlortoluron. Further, the enzyme activity assay results suggest that the mode of action of compound 3a involves the inhibition of the PPO enzyme, and 3a showed better inhibitory activity against PPO than did Flumioxazin. These results indicate that our molecular design strategy contributes to the development of novel promising PPO inhibitors.

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