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.

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.

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.

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.

Industrial applications of recombinant DNA technology

1987 ◽  
Vol 64 (4) ◽  
pp. 337 ◽  
Author(s):  
Michael D. Jones ◽  
Jeffrey T. Fayerman
Keyword(s):  
Recombinant Dna ◽  
Industrial Applications ◽  
Recombinant Dna Technology ◽  
Dna Technology

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 .

scholarly journals Microbial lipases and their applications – a review

2018 ◽  
pp. 54-76
Keyword(s):  
Recombinant Dna ◽  
Industrial Applications ◽  
Recombinant Dna Technology ◽  
Fungal Enzymes ◽  
First Choice ◽  
Medical Textile ◽  
Microbial Lipases ◽  
Increasing Demand ◽  
Key Aspects ◽  
Secretory System

This review focuses on the key aspects of lipases. Lipases (EC 3.1.1.3) are triacylglycerol acylhydrolases that act on carboxylic ester bonds. They breakdown triacylglycerides into glycerides (diglycerides or monoglycerides), fatty acids and glycerol. Their mass ranges from 19 kDa for B. stratosphericus to 92 kDa for P. gessardii. Their optimum temperature and pH ranges from 15 °C to 80 °C for Acinetobacter sp. and Janibacter sp. and 5 to 11 for P. gessardii and E. faecium respectively. Lipases chemo-, regio-, and enantio- specific features make them first choice of enzymes in research. Their kinetics for substrate hydrolysis depends on different esters. Mostly lipases are extracellular. Type 1 secretory system (T1SS) and Type 2 secretory system (T2SS) are involved in secreting lipases to external medium. They are found in eukaryotes and prokaryotes including animals, plants and microorganisms. Moreover, bacterial and fungal enzymes have diverse industrial applications in food, health, pharmaceutical, medical, textile, detergent, cosmetic and paper industries. Genetic engineering is employed to improve the properties of lipases. Their increasing demand in market has made them a hot topic in scientific research. Scientists are trying to discover novel lipase producing microorganisms due to their expanding commercial value. Keywords: Lipases, esterification, transesterification, biochemical and physicochemical properties, recombinant DNA technology

Immunogold labeling of CMP-KDO synthetase produced by recombinant E. Coli cells

1987 ◽  
Vol 45 ◽  
pp. 924-925
Author(s):  
F. A. Durum ◽  
R. G. Goldman ◽  
T. J. Bolling ◽  
M. F. Miller
Keyword(s):  
Inclusion Bodies ◽  
Large Scale ◽  
Recombinant Dna ◽  
Test System ◽  
Cell Protein ◽  
Gram Negative Bacteria ◽  
Cytoplasmic Inclusions ◽  
Isolation And Purification ◽  
E Coli ◽  
Low Concentrations

CMP-KDO synthetase (CKS) is an enzyme which plays a key role in the synthesis of LPS, an outer membrane component unique to gram negative bacteria. CKS activates KDO to CMP-KDO for incorporation into LPS. The enzyme is normally present in low concentrations (0.02% of total cell protein) which makes it difficult to perform large scale isolation and purification. Recently, the gene for CKS from E. coli was cloned and various recombinant DNA constructs overproducing CKS several thousandfold (unpublished data) were derived. Interestingly, no cytoplasmic inclusions of overproduced CKS were observed by EM (Fig. 1) which is in contrast to other reports of large proteinaceous inclusion bodies in various overproducing recombinant strains. The present immunocytochemical study was undertaken to localize CKS in these cells.Immune labeling conditions were first optimized using a previously described cell-free test system. Briefly, this involves soaking small blocks of polymerized bovine serum albumin in purified CKS antigen and subjecting them to various fixation, embedding and immunochemical conditions.

scholarly journals Dissecting cellobiose metabolic pathway and its application in biorefinery through consolidated bioprocessing in Myceliophthora thermophila

2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Jingen Li ◽  
Shuying Gu ◽  
Zhen Zhao ◽  
Bingchen Chen ◽  
Qian Liu ◽  
...  
Keyword(s):  
Malic Acid ◽  
Plant Biomass ◽  
Consolidated Bioprocessing ◽  
Cellulase Activity ◽  
Industrial Applications ◽  
Efficient Production ◽  
Myceliophthora Thermophila ◽  
Final Strain ◽  
Cellobiose Metabolism ◽  
Cost Efficient

Abstract Background Lignocellulosic biomass has long been recognized as a potential sustainable source for industrial applications. The costs associated with conversion of plant biomass to fermentable sugar represent a significant barrier to the production of cost-competitive biochemicals. Consolidated bioprocessing (CBP) is considered a potential breakthrough for achieving cost-efficient production of biomass-based fuels and commodity chemicals. During the degradation of cellulose, cellobiose (major end-product of cellulase activity) is catabolized by hydrolytic and phosphorolytic pathways in cellulolytic organisms. However, the details of the two intracellular cellobiose metabolism pathways in cellulolytic fungi remain to be uncovered. Results Using the engineered malic acid production fungal strain JG207, we demonstrated that the hydrolytic pathway by β-glucosidase and the phosphorolytic pathway by phosphorylase are both used for intracellular cellobiose metabolism in Myceliophthora thermophila, and the yield of malic acid can benefit from the energy advantages of phosphorolytic cleavage. There were obvious differences in regulation of the two cellobiose catabolic pathways depending on whether M. thermophila JG207 was grown on cellobiose or Avicel. Disruption of Mtcpp in strain JG207 led to decreased production of malic acid under cellobiose conditions, while expression levels of all three intracellular β-glucosidase genes were significantly up-regulated to rescue the impairment of the phosphorolytic pathway under Avicel conditions. When the flux of the hydrolytic pathway was reduced, we found that β-glucosidase encoded by bgl1 was the dominant enzyme in the hydrolytic pathway and deletion of bgl1 resulted in significant enhancement of protein secretion but reduction of malate production. Combining comprehensive manipulation of both cellobiose utilization pathways and enhancement of cellobiose uptake by overexpression of a cellobiose transporter, the final strain JG412Δbgl2Δbgl3 produced up to 101.2 g/L and 77.4 g/L malic acid from cellobiose and Avicel, respectively, which corresponded to respective yields of 1.35 g/g and 1.03 g/g, representing significant improvement over the starting strain JG207. Conclusions This is the first report of detailed investigation of intracellular cellobiose catabolism in cellulolytic fungus M. thermophila. These results provide insights that can be applied to industrial fungi for production of biofuels and biochemicals from cellobiose and cellulose.

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