Optimizing microbial hosts for the large-scale production of valuable metabolites often requires multiple mutations and modifications to the host's genome. We describe a three-round screen for increased L-DOPA production in S. cerevisiae using FACS enrichment of an enzyme-coupled biosensor for L-DOPA. Multiple rounds of screening were enabled by a single build of a barcoded in vitro transposon-mediated disruption library. New background strains for screening were built for each iteration using results from previous iterations. The same in vitro transposon-mediated disruption library was integrated by homologous recombination into new background strains in each round of screening. Compared with creating new transposon insertions in each round, this method takes less time and saves the cost of additional sequencing to characterize transposon insertion sites. In the first two rounds of screening, we identified deletions that improved biosensor compartmentalization and, consequently, improved our ability to screen for L-DOPA production. In a final round, we discovered that deletion of heme oxygenase (HMX1) increases total heme concentration and increases L-DOPA production, using dopamine measurement as a proxy. We further demonstrated that deleting HMX1 may represent a general strategy for P450 function improvement by improving activity of a second P450 enzyme, BM3, which performs a distinct reaction.
Download full-text PDF |
Source |
---|---|
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6456618 | PMC |
http://dx.doi.org/10.1038/s41598-019-41759-0 | DOI Listing |
Parkinsonism Relat Disord
December 2024
Roche Pharma Research and Early Development (pRED), Roche Innovation Center, F. Hoffmann-La Roche Ltd, Basel, Switzerland.
Introduction: Prasinezumab was shown to potentially delay motor progression in individuals with early-stage Parkinson's disease (PD) who were either treatment-naïve or on monoamine oxidase type B inhibitor (MAO-Bi) therapy in the PASADENA study. We report the rationale, design, and baseline patient characteristics of the PADOVA study, designed to evaluate prasinezumab in an early-stage PD population receiving standard-of-care (SOC) symptomatic medications.
Methods: PADOVA (NCT04777331) is a Phase 2b, multicenter, randomized, double-blind, placebo-controlled, parallel-group study, in which individuals with early-stage PD on SOC stable symptomatic monotherapy (levodopa or MAO-Bi) receive intravenous prasinezumab 1500 mg every 4 weeks.
Am J Geriatr Psychiatry
December 2024
HM CINAC (Centro Integral de Neurociencias Abarca Campal) (RFF, CDTP, CGS), Hospital Universitario HM Puerta del Sur, HM Hospitales. Madrid, Spain; Instituto de Investigación Sanitaria HM Hospitales (RFF, CDTP, CGS), Madrid, Spain; Network Center for Biomedical Research on Neurodegenerative Diseases (CIBERNED) (CGS), Instituto Carlos III, Madrid, Spain; University CEU-San Pablo (CGS), Madrid, Spain. Electronic address:
Parkinson's disease (PD) is a neurodegenerative disorder characterized by motor and non-motor manifestations, including alexithymia. This condition is defined by difficulty in recognizing, articulating, and expressing one's emotional states. In this study, we conducted a systematic review and meta-analysis to compare the prevalence of alexithymia in PD patients and a healthy population, and to identify associated demographic and clinical factors.
View Article and Find Full Text PDFMicrob Cell Fact
December 2024
Key Laboratory of Engineering Biology for Low-carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
Background: Dopaxanthin is a natural pigment betaxanthins family member with the highest antioxidant and free radical scavenging activities. However, its relatively low content in plants limited the wide range of applications. Cost-efficient microbial production, therefore, showed an attractive alternative.
View Article and Find Full Text PDFAnal Chem
December 2024
Department of Pharmacy, Zhongnan Hospital of Wuhan University, School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
A ratiometric fluorescence-photothermal dual-mode assay method is constructed for the detection of butyrylcholinesterase (BChE) activity based on time-resolved levodopa (L-DOPA) cascade polymerization. First, a newly designed bimetallic metal-organic framework (MOF), Eu/Co-DPA (DPA: pyridine-2,6-dicarboxylic acid), is screened out as a fluorescent nanozyme with high catalytic activity and superior luminescence properties. In the presence of boric acid (BA), L-DOPA forms BA-esterified L-DOPA, which is catalyzed by Eu/Co-DPA to form the oligomers with strong blue fluorescence.
View Article and Find Full Text PDFACS Appl Bio Mater
December 2024
Department of Materials Science and Engineering, Indian Institute of Technology Delhi, Hauz Khas, Delhi, India 110016.
Inspired by the intricate cellular morphology and the discoid shape of red blood cells (RBCs), biomimetic tricompartmental microcarriers (TCM) with controlled release profiles were engineered using an electrohydrodynamic--jetting technique for efficient management of Parkinson's disease (PD). While jetting, Levodopa (LD), CD (Carbidopa), and ENT (Entacapone) (3 PD drugs) were directly encapsulated in the three individual compartments of the TCM used for oral administration. The optimal shape and controlled release profiles were obtained by employing the Taguchi orthogonal L9 design-of-experiment approach by systematically varying the processing parameters, i.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!