Showing papers by "Qinhong Wang published in 2013"
TL;DR: A microplate‐based high‐throughput assay for the detection of succinic acid, which is widely used in many fields of industry, and the assay can also be applied to screen high succinicacid‐producing strains.
Abstract: Aims To develop a microplate-based high-throughput assay for the detection of succinic acid, which is widely used in many fields of industry. The assay can also be applied to screen high succinic acid-producing strains. Methods and Results Here, we developed a double-enzyme-based succinic acid detection assay using fumarate reductase and peroxidase (named FRDP assay) to screen and isolate high succinic acid-producing bacteria. The assay was performed in 96-well microplate in a high-throughput and had a well linear detection range of succinic acid from 0 center dot 05mmoll1 to 0 center dot 2mmoll1. Interference analysis results showed the complex media (LB),the by-products (formic acid and acetic acid) of fermentation and a longer fermentation time interfere with the FRDP assay, while the mineral media (M9) and short fermentation time (<12h) could minimize the interference. Conclusions The developed double-enzyme-coupled FRDP assay can be used in quick screening of succinic acid production. Significance and Impact of the study In this study, a novel high-throughput screening method to detect succinic acid was developed. This screening method is useful to screen succinic acid-producing bacteria and can be easily adjusted to screen different strains for the production of many other metabolites.
10 citations
TL;DR: One mutant from 792 clones showed specific substrate activity of converting progesterone to 16,17-epoxysterol, which validated the possibility to evolve the cytochrome P450 for the synthesis of steroidal epoxides.
Abstract: 16,17-Epoxysterol plays an important role in pharmaceutical steroid synthesis. To investigate the potential application of cytochrome P450 for epoxysterol synthesis, an approach to the epoxidation of 16,17-epoxysterol, based on directed evolution of cytochrome P450 BM-3, was developed. This comprised random gene mutagenesis for optimizing the activity of P450 BM-3 for epoxidation of hydrophobic sterol, followed by the 7-ethoxycoumarin de-ethylation assay for general enzyme activity detection and the modified picric acid assay for epoxidation activity screening. By the two-step screening, one mutant from 792 clones showed specific substrate activity of converting progesterone to 16,17-epoxysterol, which validated the possibility to evolve the cytochrome P450 for the synthesis of steroidal epoxides.
9 citations
Journal Article•
TL;DR: The research advances of adipic acid and its precursor production are reviewed, followed by addressing the perspective of the possible new pathways for adipic Acid production.
6 citations
Journal Article•
TL;DR: A new 88 bp synthetic promoter, which contains the typical -35 box, -10 box as well as ribosome bind site, was designed and the results showed that the promoters with varying strengths can regulate the production of cis,cis-muconic acid and the accumulation of the intermediate catechol.
5 citations
TL;DR: As a steroidal drug intermediate, 16,17-epoxysterol plays an important role in pharmaceutical steroid synthesis and its determination normally require expensive equipment, particularly for high-performance liquid chromatography.
Abstract: As a steroidal drug intermediate, 16,17-epoxysterol plays an important role in pharmaceutical steroid synthesis. Current methods for epoxysterol determination normally require expensive equipment, ...