Enzymatic Synthesis of 15N-L-aspartic Acid Using Recombinant Aspartase from Escherichia Coli K12

2008 ◽  
Vol 59 (11) ◽  
Author(s):  
Iulia Lupan ◽  
Sergiu Chira ◽  
Maria Chiriac ◽  
Nicolae Palibroda ◽  
Octavian Popescu
Keyword(s):  
Amino Acids ◽  
Aspartic Acid ◽  
Enzymatic Synthesis ◽  
Large Scale ◽  
Recombinant Dna ◽  
Industrial Enzymes ◽  
Recombinant Dna Technology ◽  
Bacterial Fermentation ◽  
Natural Protein ◽  
Novel Method

Amino acids are obtained by bacterial fermentation, extraction from natural protein or enzymatic synthesis from specific substrates. With the introduction of recombinant DNA technology, it has become possible to apply more rational approaches to enzymatic synthesis of amino acids. Aspartase (L-aspartate ammonia-lyase) catalyzes the reversible deamination of L-aspartic acid to yield fumaric acid and ammonia. It is one of the most important industrial enzymes used to produce L-aspartic acid on a large scale. Here we described a novel method for [15N] L-aspartic synthesis from fumarate and ammonia (15NH4Cl) using a recombinant aspartase.

scholarly journals Development Of A Gradient System For The Establishment Of Cellulolytic Microbial Communities

2021 ◽  
Author(s):  
Ashley Sousa
Keyword(s):  
Large Scale ◽  
Recombinant Dna ◽  
Consolidated Bioprocessing ◽  
Environmental Gradients ◽  
Industrial Applications ◽  
Single Step ◽  
Gradient System ◽  
Recombinant Dna Technology ◽  
Selective Enrichment ◽  
Feasible Alternative

Cellulosic ethanol has shown promise as a feasible alternative fuel, especially if the hydrolysis of lignocellulosic biomass is done through a single step process known as consolidated bioprocessing (CBP). A major challenge for CBP, especially for large-scale industrial applications is the inhibition of celluloytic microorganisms by ethanol. While recombinant DNA technology and microbial acclimatization by exposure have resulted in some increase in ethanol tolerance, the search remains for robust bacteria that can proliferate in industrially-relevant conditions. This study applied an anaerobic gradient system to provide a continous spatial pathway for the selection of cellulolytic consortia with increased tolerance to ethanol. DGGE analysis showed that increasing concentrations of ethanol impacts the community profile. Biofilm formation of cellulose degrading communities has been found to be influenced by species diversity. Environmental gradients have shown promise for selective enrichment of cellulolytic consortia at desired conditions required for industrial application.

Rapid large-scale purification of plasmid DNA by medium or low pressure gel filtration. Application: construction of thermoamplifiable expression vectors

1985 ◽  
Vol 5 (2) ◽  
pp. 101-111 ◽  
Author(s):  
Tuyen Vo-Quang ◽  
Yves Malpiece ◽  
Dominique Buffard ◽  
P. Alexandre Kaminski ◽  
Dominique Vidal ◽  
...  
Keyword(s):  
Large Scale ◽  
Recombinant Dna ◽  
Gel Filtration ◽  
Low Pressure ◽  
Expression Vectors ◽  
Recombinant Dna Technology ◽  
Hybrid Proteins ◽  
Protein Contamination ◽  
As Adsorption ◽  
Foreign Genes

This paper describes a new method of lasmid DNA purification which is fast and reliable enough for most purposes in recombinant DNA technology. The present method does not require the use of toxic chemicals such as phenol or ethidium bromide, costly ultra-centrifugation procedures or other processes which can modify the supercoiled structure of the plasmids, such as adsorption on glass fiber. This method is based on the principle of gel filtration chromatography, at low pressure (1 bar) or medium pressure (between 5 and 10 bars), using Sephacryl S1000 or Superose 6B. It permits recovery oI plasmids: (I) in preparative quantities (from 300 gg to 4 mg), (II) exempt from RNA, DNA and protein contamination, and (III) suitable for various common genetic engineering procedures immediately after purification. To test the reliability of the technique as well as the degree of purilication, the plasmids were used to construct thermoampliIiable vectors, carrying the tacUV5 promoter and the 5′ end of the β -gallactosidase gone with a single EcoRl site in each of the three possible translational phases. This set of vectors is designed for the expression of foreign genes as hybrid proteins in Escherichia coli.

scholarly journals Development Of A Gradient System For The Establishment Of Cellulolytic Microbial Communities

2021 ◽  
Author(s):  
Ashley Sousa
Keyword(s):  
Large Scale ◽  
Recombinant Dna ◽  
Consolidated Bioprocessing ◽  
Environmental Gradients ◽  
Industrial Applications ◽  
Single Step ◽  
Gradient System ◽  
Recombinant Dna Technology ◽  
Selective Enrichment ◽  
Feasible Alternative

Cellulosic ethanol has shown promise as a feasible alternative fuel, especially if the hydrolysis of lignocellulosic biomass is done through a single step process known as consolidated bioprocessing (CBP). A major challenge for CBP, especially for large-scale industrial applications is the inhibition of celluloytic microorganisms by ethanol. While recombinant DNA technology and microbial acclimatization by exposure have resulted in some increase in ethanol tolerance, the search remains for robust bacteria that can proliferate in industrially-relevant conditions. This study applied an anaerobic gradient system to provide a continous spatial pathway for the selection of cellulolytic consortia with increased tolerance to ethanol. DGGE analysis showed that increasing concentrations of ethanol impacts the community profile. Biofilm formation of cellulose degrading communities has been found to be influenced by species diversity. Environmental gradients have shown promise for selective enrichment of cellulolytic consortia at desired conditions required for industrial application.

Biosynthesis and Processing of Silk Proteins

MRS Bulletin ◽  
1992 ◽  
Vol 17 (10) ◽  
pp. 41-47 ◽  
Author(s):  
David L. Kaplan ◽  
Stephen Fossey ◽  
Charlene M. Mello ◽  
Steven Arcidiacono ◽  
Kris Senecal ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Recombinant Dna ◽  
Hydrophobic Interactions ◽  
Genetic Manipulation ◽  
Larval Instar ◽  
Side Chain ◽  
Recombinant Dna Technology ◽  
Short Side ◽  
Orb Web

Silks produced by silkworms (e.g., Bombyx mori) and orb-web weaving spiders (e.g., Nephila clavipes) are essentially pure protein, that is, complexes of amino acid polymers. They are the most common fibers spun by biological systems. There has been a long-standing interest in the use of these and similar fibers in textiles, cables, fiber reinforcement in composites, in addition, for example, to cross hairs in optical instruments, and fishing nets. Both nylon, a homo-polymer of the amino acid glycine, and Kevlar, a polymer of a nonnatural aromatic amino acid, can be considered modified, synthetic versions of silk and are used for some of the applications mentioned above. The potential for genetic manipulation, through recombinant DNA technology, of the natural biosynthetic process for these natural proteins (see the article by Cappello in this issue) has renewed interest in the production of new silklike proteins.The natural silks are characterized by a β-sheet secondary structure which is stabilized by interchain hydrogen bonds and intersheet hydrophobic interactions (Figure 1). Silks can be considered block copolymers, with crystalline domains consisting of short side chain monomers (the amino acids glycine, alanine, and serine) interspersed in amorphous domains consisting of bulkier side chain amino acids.This family of fibers is naturally tailored to perform functions such as catching prey (orb web) or serving as a barrier against environmental challenges (cocoon). The domestic silkworm (B. mori) produces only one type of silk, cocoon silk, at only one stage in its lifecyle, during the fifth larval instar just before molt to the pupa. The silk is produced in modified salivary glands and spun from the mouth.

scholarly journals 44 Amino acids in livestock: A historical perspective

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 22-22
Author(s):  
Keith D Haydon ◽  
R Dean Boyd
Keyword(s):  
Amino Acids ◽  
Chemical Synthesis ◽  
Production Cost ◽  
Recombinant Dna ◽  
Cost Effective ◽  
Commercial Application ◽  
Recombinant Dna Technology ◽  
Umami Taste ◽  
Bacterial Mutants ◽  
Poultry Diets

Abstract The advent of commercially viable synthetic amino acids (AA) and least cost formulation (LCF) have fundamentally changed swine and poultry diets over the past 40 years. Amino acids can be produced by chemical synthesis, hydrolysis of intact proteins and fermentation. Chemical synthesis of AA was first reported by Strecker (1850). Discovery of glutamate as the basis of Umami taste category (1907) spurred commercial AA production by hydrolysis. The discovery of commercially viable fermentative production of glutamate, by Kyowa Hakko Kogyo Co. (1957), revolutionized AA production. Their parallel discovery of lysine (Lys) production, using a natural C. glutmanicum mutant subsequently followed. This led to further screening for bacterial mutants for threonine (Thr, 1961), tryptophan (Trp, 1972), valine (Val, 1959) and isoleucine (Ile, 1972). The next milestone occurred with development of the first main-frame LCF programs (1960s). The first commercial application of dietary AA involved methionine (Met) for poultry, followed closely by Lys in swine. Use of other synthetic AA was cost-prohibitive, but they served as research tools. The next revolution involved recombinant DNA technology (1980s), which dramatically increased AA yield and reduced production cost. Simultaneously, development of PC based LCF enabled cost effective formulation. Subsequently, growth-derived AA ratio’s emerged from the labs of Fuller (1989) and Baker (1992). Patent expirations in the late 1980s led to new companies that produced rapid advances in fermentation methods, with new recombinant strains. Production cost declined further for Lys (1988), and production efficiencies allowed Thr (1995) and Trp (2000) to enter commercial diets. Advances in fermentation technology have enabled production of all ten essential AA. Extensive AA displacement of protein supplements has led to an ever-expanding global tonnage of AA for food and pet animals. With routine addition of 4–6 AA in swine diets, we question whether non-essential AA nitrogen may emerge as limiting (essential).

scholarly journals Enzymes of industrial interest

2017 ◽  
Vol 2 (2) ◽  
pp. 74-97 ◽  
Author(s):  
Arnold L. Demain ◽  
Sergio Sánchez
Keyword(s):  
Recombinant Dna ◽  
Animal Feed ◽  
Strain Improvement ◽  
Major Effect ◽  
Industrial Enzymes ◽  
Recombinant Enzymes ◽  
Recombinant Dna Technology ◽  
Microbial Enzymes ◽  
Wide Range ◽  
The World

For many years, industrial enzymes have played an important role in the benefit of our society due to their many useful properties and a wide range of applications. They are key elements in the progress of many industries including foods, beverages, pharmaceuticals, diagnostics, therapy, personal care, animal feed, detergents, pulp and paper, textiles, leather, chemicals and biofuels. During recent decades, microbial enzymes have replaced many plant and animal enzymes. This is because microbial enzymes are widely available and produced economically in short fermentations and inexpensive media. Screening is simple, and strain improvement for increased production has been very successful. The advances in recombinant DNA technology have had a major effect on production levels of enzymes and represent a way to overproduce industrially important microbial, plant and animal enzymes. It has been calculated that 50-60% of the world enzyme market is supplied with recombinant enzymes. Molecular methods, including genomics and metagenomics, are being used for the discovery of new enzymes from microbes. Also, directed evolution has allowed the design of enzyme specificities and better performance.

Based on DNA OTP Key Generation and Management Research

2013 ◽  
Vol 427-429 ◽  
pp. 2470-2472
Author(s):  
Yun Peng Zhang ◽  
Feng Ying Tian ◽  
Man Hui Sun ◽  
Ding Yu ◽  
Fei Xiang Fan ◽  
...  
Keyword(s):  
Data Storage ◽  
Technology Use ◽  
Recombinant Plasmid ◽  
Large Scale ◽  
Recombinant Dna ◽  
Restriction Enzymes ◽  
Key Generation ◽  
Recombinant Dna Technology ◽  
New Approach ◽  
Large Scale Data

With the development of molecular-bio technology, the feature of DNA molecules for ultra-large-scale data storage has created a new approach for data storage. This paper gives a way of strengthening key transport security. Through recombinant DNA technology, use only sender-receiver know restriction enzymes to combine the key DNA and the T vector, to form a recombinant plasmid, making the key DNA bio-hide, and then place the recombinant plasmid in implanted bacteria .

Synthesis of Fullerene-Acid Conjugates

2012 ◽  
Vol 463-464 ◽  
pp. 538-542 ◽  
Author(s):  
Jing Zhang ◽  
Li Yuan ◽  
Ya Dong Zhang
Keyword(s):  
Amino Acids ◽  
Glutamic Acid ◽  
Aspartic Acid ◽  
Azomethine Ylide ◽  
Hydroxyl Group ◽  
Dipolar Cycloaddition ◽  
Chemical Structures ◽  
Ft Ir ◽  
Novel Method ◽  
Potential Use

N-substituted 3,4-fullero pyrrolidine was synthesized according to 1,3-Dipolar cycloaddition of the azomethine ylide. Aspartic acid and glutamic acid with protected α-amino and α-carboxyl groups were reacted with the activated hydroxyl group of N-substituted 3,4-fullero pyrrolidine, respectively. The products were deprotected, affording two novel fullerene α-amino acids, fullerene aspartic acid and fullerene glutamic acid. Their chemical structures were characterized by MALAI-TOF-MS, UV-Vis, FT-IR and 1HNMR. Both fullerene amino acids with a free amino group and a free carboxyl group would have unique property and potential use in medicine and biology. A novel method has been developed to synthesize fullerene conjugate. Their unique chemical structures make them very interesting for their potential use in medicine and biology.

Sign in / Sign up

Export Citation Format

Share Document