ArticlePloS one2013
Statistical experimental design guided optimization of a one-pot biphasic multienzyme total synthesis of amorpha-4,11-diene.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
22 citing papers in PubMed.
- The Mevalonate Pathway: Innovations, Applications, and Challenges in Biotechnology with Emphasis on Fungal Biology.Journal of fungi (Basel, Switzerland) · 2026Review
- EnzRetro: Enzymatic Retrosynthetic Planning With Site-Specific Reaction Edits Based on Sequence Generative Architecture.Exploration (Beijing, China) · 2026Article
- Reaction Engineering of In Vitro Natural Product Biosynthesis: Challenges and Strategies.Chembiochem : a European journal of chemical biology · 2025Review
- Enhancing Multi-Enzyme Cascade Activity in Metal-Organic Frameworks via Controlled Enzyme Encapsulation.Small (Weinheim an der Bergstrasse, Germany) · 2025Article
- Cell-Free Reaction System for ATP Regeneration from d-Fructose.ACS synthetic biology · 2025Article
- Biochemical synthesis of taxanes from mevalonate.Synthetic and systems biotechnology · 2024Article
- Quantitative Online Monitoring of an Immobilized Enzymatic Network by Ion Mobility-Mass Spectrometry.Journal of the American Chemical Society · 2024Article
- Alternative design strategies to help build the enzymatic retrosynthesis toolbox.RSC chemical biology · 2022Article
- Industrially Relevant Enzyme Cascades for Drug Synthesis and Their Ecological Assessment.International journal of molecular sciences · 2022Review
- Amorpha-4,11-diene synthase: a key enzyme in artemisinin biosynthesis and engineering.aBIOTECH · 2021Review
- A combined experimental and modelling approach for the Weimberg pathway optimisation.Nature communications · 2020Article
- A critical comparison of cellular and cell-free bioproduction systems.Current opinion in biotechnology · 2019Review
- Synthetic Biology Goes Cell-Free.BMC biology · 2019Review
- Cell-free biosynthesis of limonene using enzyme-enrichedSynthetic biology (Oxford, England) · 2019Article
- Expanding the boundary of biocatalysis: design and optimization of in vitro tandem catalytic reactions for biochemical production.Critical reviews in biochemistry and molecular biology · 2018Review
- Synthetic microbial consortia enable rapid assembly of pure translation machinery.Nature chemical biology · 2018Article
- Multienzyme Biosynthesis of Dihydroartemisinic Acid.Molecules (Basel, Switzerland) · 2017Article
- Cross-Regulation of an Artificial Metalloenzyme.Angewandte Chemie (International ed. in English) · 2017Article
- Modules forSynthetic and systems biotechnology · 2017Review
- Cell-Free Mixing of Escherichia coli Crude Extracts to Prototype and Rationally Engineer High-Titer Mevalonate Synthesis.ACS synthetic biology · 2016Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In vitro synthesis of chemicals and pharmaceuticals using enzymes is of considerable interest as these biocatalysts facilitate a wide variety of reactions under mild conditions with excellent regio-, chemo- and stereoselectivities. A significant challenge in a multi-enzymatic reaction is the need to optimize the various steps involved simultaneously so as to obtain high-yield of a product. In this study, statistical experimental design was used to guide the optimization of a total synthesis of amorpha-4,11-diene (AD) using multienzymes in the mevalonate pathway. A combinatorial approach guided by Taguchi orthogonal array design identified the local optimum enzymatic activity ratio for Erg12:Erg8:Erg19:Idi:IspA to be 100∶100∶1∶25∶5, with a constant concentration of amorpha-4,11-diene synthase (Ads, 100 mg/L). The model also identified an unexpected inhibitory effect of farnesyl pyrophosphate synthase (IspA), where the activity was negatively correlated with AD yield. This was due to the precipitation of farnesyl pyrophosphate (FPP), the product of IspA. Response surface methodology was then used to optimize IspA and Ads activities simultaneously so as to minimize the accumulation of FPP and the result showed that Ads to be a critical factor. By increasing the concentration of Ads, a complete conversion (∼100%) of mevalonic acid (MVA) to AD was achieved. Monovalent ions and pH were effective means of enhancing the specific Ads activity and specific AD yield significantly. The results from this study represent the first in vitro reconstitution of the mevalonate pathway for the production of an isoprenoid and the approaches developed herein may be used to produce other isopentenyl pyrophosphate (IPP)/dimethylallyl pyrophosphate (DMAPP) based products.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.