Metabolic Fingerprint Analysis of Cytochrome -producing N4830-1 Using FT-IR Spectroscopy.

Front Microbiol

Centre for Metabolomics Research, Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom.

Published: June 2022

Optimization of recombinant protein expression in bacteria is an important task in order to increase protein yield while maintaining the structural fidelity of the product. In this study, we employ Fourier transform infrared (FT-IR) spectroscopy as a high throughput metabolic fingerprinting approach to optimize and monitor cytochrome (CYT ) production in N4830-1, as the heterologous host. Cyt b was introduced as a plasmid with between 0 and 6 copies under a strong promoter. The FT-IR spectroscopy results combined with multivariate chemometric analysis illustrated discriminations among culture conditions as well as revealing features that correlated to the different gene copy numbers. The second derivative of the FT-IR spectral data allowed for the quantitative detection of Cyt b directly inside the intact cells without the need for extraction, and highlighted changes in protein secondary structure that was directly correlated to the gene copy number and protein content, and was in complete agreement with quantitative findings of standard traditional techniques such as SDS-PAGE and western blot analysis.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9257212PMC
http://dx.doi.org/10.3389/fmicb.2022.874247DOI Listing

Publication Analysis

Top Keywords

ft-ir spectroscopy
12
correlated gene
8
gene copy
8
metabolic fingerprint
4
fingerprint analysis
4
analysis cytochrome
4
cytochrome -producing
4
-producing n4830-1
4
ft-ir
4
n4830-1 ft-ir
4

Similar Publications

In this study, an optical sensor, JA/(2,6-di((E)-benzylidene)cyclohexan-1-one), was synthesized and characterized using H NMR and FT-IR spectroscopy. The sensor exhibited high efficiency and selectivity in detecting Pb ions, even in the presence of potential interfering ions such as Mn, Cu, Co, Cr, Ni, Ce, Hg, and Cd in aqueous solutions. The interaction of JA with Pb resulted in a significant enhancement of fluorescence intensity, suggesting the formation of a stable complex.

View Article and Find Full Text PDF

Objectives: This study focuses on both the formulation of bio-based microspheres containing fampridine for the treatment of multiple sclerosis and provides an alternative to the commercially available product (Fampyra 10 mg, Biogen).

Materials And Methods: The encapsulation of fampridine was achieved using polyvinyl alcohol (PVA) and sodium alginate (Na-Alg) polymers. Glutaraldehyde (GA) and hydrochloric acid (HCI) were used as crosslinking agents.

View Article and Find Full Text PDF

The importance of developing multifunctional nanomaterials for sensing technologies is increasing with the arrival of nanotechnology. In this study, we describe the introduction of novel nanoprobe electro-active material into the architecture of an electrochemical immuno-sensor. Based on the electrochemical immuno-sensor, functionalized tin oxide/graphitic carbon nitride nanocomposite (fSnO/g-CN) was synthesized and then analyte specific anti-aflatoxin M monoclonal antibody (AFM-ab) combined to form an electro-active nanoprobe (fSnO/g-CN/AFM-ab).

View Article and Find Full Text PDF

Nitrogen doped Carbon Quantum Dots (NCQDs) have been synthesized using most economical and easiest hydrothermal process. Here, N-phenyl orthophenylenediamine and citric acid were utilised as a source of nitrogen and carbon for the preparation of NCQDs. The synthesized NCQDs were characterized using experimental techniques like UV - Vis absorption, FT-IR, transmission electron microscopy (TEM), X-ray Diffraction (XRD), EDX, dynamic light scattering (DLS), fluorimeter and time resolved fluorescence spectroscopy.

View Article and Find Full Text PDF

The Properties of Damaged Starch Granules: The Relationship Between Granule Structure and Water-Starch Polymer Interactions.

Foods

December 2024

Instituto de Ciencia y Tecnología de los Alimentos Córdoba (ICYTAC-CONICET), Universidad Nacional de Córdoba, Av. Filloy S/N, Ciudad Universitaria, Córdoba CP 5000, Argentina.

The morphology of wheat starch granules with different damaged starch (DS) content was analyzed using a particle size analyzer and scanning electron microscopy (SEM); the granular structure was studied using FT-IR spectroscopy and X-ray diffraction (XRD); and the granule-water interaction was evaluated by thermogravimetric analysis (TGA) and dynamic vapor sorption (DVS). The increase in the level of DS shifted the population of B-type granules towards larger particle diameters and shifted the population of A-type granules towards smaller particle diameters. The appearance of the surface of the starch-damaged granules was rough and flaky (SEM images).

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!