Topics

The Role of Amino Acids in the Amplification and Quality of DNA Vectors for Industrial Applications.

08:00 EDT 12th July 2019 | BioPortfolio

Summary of "The Role of Amino Acids in the Amplification and Quality of DNA Vectors for Industrial Applications."

In this study, we have demonstrated that the type and feeding regimen of amino acids have a significant impact on the quality as well as the quantity of DNA vectors produced. Nutrient pool and factorial design experiments were carried out in order to identify the amino acids involved in increased biomass and induction of plasmid amplification. Leucine, glycine, and histidine were responsible for increased biomass and leucine starvation in the presence of histidine was implicated in plasmid amplification. Supercoiling of the plasmid was optimised using a dual feeding strategy. As a result of this, a fed-batch fermentation strategy for the production of a 6.9kb plasmid, pSVß, in Escherichia coli DH5α was developed. In batch fermentation, a maximum plasmid yield of 39.4mg/L equivalent to 11.3mg/g DCW was achieved with casein hydrolysate limitation. 90% of plasmid was in the supercoiled form after 31h of fermentation but only remained so for a short period, leading to a very brief window for harvesting cells at scale. Subsequently, a fed-batch fermentation using a dual feeding strategy was employed. A mean maximum plasmid yield of 44 mg/L equivalent to 9.1 mg plasmid/g DCW was achieved. After 25h, 90% of plasmid was in the supercoiled form and remained at this level for the remaining 10h of the fermentation, allowing adequate time for the harvesting of cells without the loss of supercoiling of product. This study emphasized that optimizing fermentation strategy and identifying the essential nutrients are beneficial for bioprocessing of plasmid DNA for therapeutic applications. This article is protected by copyright. All rights reserved.

Affiliation

Journal Details

This article was published in the following journal.

Name: Biotechnology progress
ISSN: 1520-6033
Pages: e2883

Links

DeepDyve research library

PubMed Articles [27408 Associated PubMed Articles listed on BioPortfolio]

A Model of Molecular Vector Machine of Proteins.

Based on the analysis of NH….O=C bond region of the proteins pentafragments using the quantum chemistry representations, a group of 20 vectors of action has been isolated. These vectors of action ar...

Prostate Cancer-specific Hallmarks of Amino Acids Metabolism: Towards a Paradigm of Precision Medicine.

So far multiple differences in prostate cancer-specific amino acids metabolism have been discovered. Moreover, some attempts to utilize these alterations for prostate cancer diagnosis and treatment ha...

N-Alkyl-α-amino acids in Nature and their biocatalytic preparation.

N-alkylated-α-amino acids are useful building blocks for the pharmaceutical and fine chemical industries. Enantioselective methods of N-alkylated-α-amino acid synthesis are therefore highly valuable...

Free and combined L- and D-amino acids in Arctic aerosol.

Aerosol samples were collected with a high-volume cascade impactor with a 10 day sampling frequency at the Gruvebadet observatory, close to Ny-Ålesund (Svalbard Islands). A total of 42 filters were a...

Objective and Subjective Speech Quality Assessment of Amplification Devices for Patients with Parkinson's Disease.

This paper investigated subjective and objective assessment of Parkinsonian speech quality. Speech stimuli were recorded from 11 Parkinsonian and 10 age-matched normal control participants under diffe...

Clinical Trials [6588 Associated Clinical Trials listed on BioPortfolio]

Effect of Non-essential Amino Acids on Protein Requirements for Endurance Athletes

Protein requirements in individuals who participate in endurance-based exercise training have been suggested to be greater than the current recommended dietary allowance (RDA). The biolog...

Effect of Branched Chain Amino Acids on Muscle

With aging, there is a decrease in muscle mass and function especially in the energy storehouses of cells called mitochondria. Amino acids, the building blocks of protein, and insulin hav...

Dynamic Aspects of Amino Acid Metabolism

This protocol seeks to define aspects of intestinal and hepatic uptake and metabolism of several amino acids. The major hypothesis to be tested is that the splanchic bed (intestine and li...

Amino Acids in Urine, Diet, and Blood Pressure: International Population Study

To analyze the relationship of dietary variables to urinary excretion of amino acids and the relationships of specific urinary amino acids to blood pressure.

Preoperative Oral Supplementation With Branched-Chain Amino Acids (BCAA) Improves Postoperative Quality of Life

The aims of this study are (1) to study the benefit of branched-chain amino acids enriched nutrient-mixture by analyzing a variety of clinical parameter, including laboratory data, the dur...

Medical and Biotech [MESH] Definitions

Oils which are used in industrial or commercial applications.

Amino acids and chains of amino acids connected by peptide linkages.

The application of industrial management practice to systematically maintain and improve organization-wide performance. Effectiveness and success are determined and assessed by quantitative quality measures.

Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are linear polypeptides that are normally synthesized on RIBOSOMES.

Endogenous amino acids released by neurons as excitatory neurotransmitters. Glutamic acid is the most common excitatory neurotransmitter in the brain. Aspartic acid has been regarded as an excitatory transmitter for many years, but the extent of its role as a transmitter is unclear.

Quick Search


DeepDyve research library

Relevant Topics

Bioinformatics
Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...

Nutrition
Within medicine, nutrition (the study of food and the effect of its components on the body) has many different roles. Appropriate nutrition can help prevent certain diseases, or treat others. In critically ill patients, artificial feeding by tubes need t...


Searches Linking to this Article