scholarly journals From Data Mining of Chitinophaga sp. Genome to Enzyme Discovery of a Hyperthermophilic Metallocarboxypeptidase

2021 ◽  
Vol 9 (2) ◽  
pp. 393
Author(s):  
Gabriela Cabral Fernandes ◽  
Elwi Guillermo Machado Sierra ◽  
Paul Brear ◽  
Mariana Rangel Pereira ◽  
Eliana G. M. Lemos
Keyword(s):  
Data Mining ◽  
Bacterial Consortium ◽  
Industrial Applications ◽  
Biotechnological Applications ◽  
Negative Bacterium ◽  
Conserved Motif ◽  
E Coli ◽  
Enzyme Discovery ◽  
Ethanol Plant ◽  
Positive Effect

For several centuries, microorganisms and enzymes have been used for many different applications. Although many enzymes with industrial applications have already been reported, different screening technologies, methods and approaches are constantly being developed in order to allow the identification of enzymes with even more interesting applications. In our work, we have performed data mining on the Chitinophaga sp. genome, a gram-negative bacterium isolated from a bacterial consortium of sugarcane bagasse isolated from an ethanol plant. The analysis of 8 Mb allowed the identification of the chtcp gene, previously annotated as putative Cht4039. The corresponding codified enzyme, denominated as ChtCP, showed the HEXXH conserved motif of family M32 from thermostable carboxypeptidases. After expression in E. coli, the recombinant enzyme was characterized biochemically. ChtCP showed the highest activity versus benziloxicarbonil Ala-Trp at pH 7.5, suggesting a preference for hydrophobic substrates. Surprisingly, the highest activity of ChtCP observed was between 55 °C and 75 °C, and 62% activity was still displayed at 100 °C. We observed that Ca2+, Ba2+, Mn2+ and Mg2+ ions had a positive effect on the activity of ChtCP, and an increase of 30 °C in the melting temperature was observed in the presence of Co2+. These features together with the structure of ChtCP at 1.2 Å highlight the relevance of ChtCP for further biotechnological applications.

Enhanced Biocatalytic Activity of Recombinant Lipase Immobilized on Gold Nanoparticles

2019 ◽  
Vol 20 (6) ◽  
pp. 497-505 ◽  
Author(s):  
Abeer M. Abd El-Aziz ◽  
Mohamed A. Shaker ◽  
Mona I. Shaaban
Keyword(s):  
Gold Nanoparticles ◽  
Lipolytic Activity ◽  
Market Demand ◽  
Biotechnological Applications ◽  
Lipase Gene ◽  
Expression Plasmid ◽  
E Coli ◽  
Nickel Affinity Chromatography ◽  
Recombinant Production ◽  
Sds Page

Background: Bacterial lipases especially Pseudomonas lipases are extensively used for different biotechnological applications. Objectives: With the better understanding and progressive needs for improving its activity in accordance with the growing market demand, we aimed in this study to improve the recombinant production and biocatalytic activity of lipases via surface conjugation on gold nanoparticles. Methods: The full length coding sequences of lipase gene (lipA), lipase specific foldase gene (lipf) and dual cassette (lipAf) gene were amplified from the genomic DNA of Pseudomonas aeruginosa PA14 and cloned into the bacterial expression vector pRSET-B. Recombinant lipases were expressed in E. coli BL-21 (DE3) pLysS then purified using nickel affinity chromatography and the protein identity was confirmed using SDS-PAGE and Western blot analysis. The purified recombinant lipases were immobilized through surface conjugation with gold nanoparticles and enzymatic activity was colorimetrically quantified. Results: Here, two single expression plasmid systems pRSET-B-lipA and pRSET-B-lipf and one dual cassette expression plasmid system pRSET-B-lipAf were successfully constructed. The lipolytic activities of recombinant lipases LipA, Lipf and LipAf were 4870, 426 and 6740 IUmg-1, respectively. However, upon immobilization of these recombinant lipases on prepared gold nanoparticles (GNPs), the activities were 7417, 822 and 13035 IUmg-1, for LipA-GNPs, Lipf-GNPs and LipAf-GNPs, respectively. The activities after immobilization have been increased 1.52 and 1.93 -fold for LipA and LipAf, respectively. Conclusion: The lipolytic activity of recombinant lipases in the bioconjugate was significantly increased relative to the free recombinant enzyme where immobilization had made the enzyme attain its optimum performance.

P. vortex-mediated strategies for polysaccharides decomposition

TECHNOLOGY ◽  
2015 ◽  
Vol 03 (02n03) ◽  
pp. 80-83
Author(s):  
Mark Polikovsky ◽  
Eshel Ben-Jacob ◽  
Alin Finkelshtein
Keyword(s):  
Degradation Products ◽  
Cellulose Degradation ◽  
Cellulose Hydrolysis ◽  
Industrial Applications ◽  
Biofuel Production ◽  
E Coli ◽  
Novel Approach ◽  
Textile Manufacture ◽  
Hydrolysis Of Cellulose ◽  
The Relationship

Cellulose hydrolysis has many industrial applications such as biofuel production, food, paper and textile manufacture. Here, we present a novel approach to cellulose hydrolysis using a consortium of motile bacteria, Paenibacillus vortex, that can swarm on solid medium carrying a non-motile recombinant E. coli cargo strain expressing the β-glucosidase and cellulase genes that facilitate the hydrolysis of cellulose. These two species cooperate; the relationship is mutually beneficial: the E. coli is dispersed over long distances, while the P. vortex bacteria gain from the supply of cellulose degradation products. This enables the use of such consortia in this area of biotechnology.

scholarly journals The Response of Corn Acreage to Ethanol Plant Siting

2014 ◽  
Vol 46 (2) ◽  
pp. 157-171 ◽  
Author(s):  
Yehushua Shay Fatal ◽  
Walter N. Thurman
Keyword(s):  
Ethanol Production ◽  
Production Capacity ◽  
County Level ◽  
Corn Production ◽  
Ethanol Plant ◽  
Annual Changes ◽  
Positive Effect

U.S. ethanol production capacity increased more than threefold between 2002 and 2008. We study the effect of this growth on corn acreage. Connecting annual changes in county-level corn acreage to changes in ethanol plant capacities, we find a positive effect on planted corn. The building of a typical plant is estimated to increase corn in the county by over 500 acres and to increase acreage in surrounding counties up to almost 300 miles away. All ethanol plants are estimated to increase corn production by less than their annual requirements.

scholarly journals Recent Developments in Production and Biotechnological Applications of C-Phycocyanin

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
M. Kuddus ◽  
P. Singh ◽  
G. Thomas ◽  
Awdah Al-Hazimi
Keyword(s):  
Industrial Applications ◽  
Biotechnological Applications ◽  
Synthetic Compounds ◽  
Natural Pigments ◽  
Extraction And Purification ◽  
Natural Colorants ◽  
Extensive Range ◽  
Recent Developments ◽  
Pharmaceutical Industries ◽  
Harmful Effects

An extensive range of pigments including phycobiliproteins are present in algae. C-phycocyanin (C-PC), a phycobiliprotein, is one of the key pigments ofSpirulina, a microalgae used in many countries as a dietary supplement. Algal pigments have massive commercial value as natural colorants in nutraceutical, cosmetics, and pharmaceutical industries, besides their health benefits. At present, increasing awareness of harmful effects of synthetic compounds and inclination of community towards the usage of natural products have led to the exploitation of microalgae as a source of natural pigments/colors. This review describes recent findings about the sources and production of C-PC, with emphasis on specific techniques for extraction and purification, along with potential industrial applications in diagnostics, foods, cosmetics, and pharmaceutical industries.

scholarly journals Microalgal Enzymes with Biotechnological Applications

Marine Drugs ◽  
2019 ◽  
Vol 17 (8) ◽  
pp. 459 ◽  
Author(s):  
Giorgio Maria Vingiani ◽  
Pasquale De Luca ◽  
Adrianna Ianora ◽  
Alan D.W. Dobson ◽  
Chiara Lauritano
Keyword(s):  
State Of The Art ◽  
Industrial Applications ◽  
Biofuel Production ◽  
Biotechnological Applications ◽  
Efficient Manner ◽  
Environmentally Sustainable ◽  
Essential Components ◽  
Cost Efficient ◽  
Number Of Publications

Enzymes are essential components of biological reactions and play important roles in the scaling and optimization of many industrial processes. Due to the growing commercial demand for new and more efficient enzymes to help further optimize these processes, many studies are now focusing their attention on more renewable and environmentally sustainable sources for the production of these enzymes. Microalgae are very promising from this perspective since they can be cultivated in photobioreactors, allowing the production of high biomass levels in a cost-efficient manner. This is reflected in the increased number of publications in this area, especially in the use of microalgae as a source of novel enzymes. In particular, various microalgal enzymes with different industrial applications (e.g., lipids and biofuel production, healthcare, and bioremediation) have been studied to date, and the modification of enzymatic sequences involved in lipid and carotenoid production has resulted in promising results. However, the entire biosynthetic pathways/systems leading to synthesis of potentially important bioactive compounds have in many cases yet to be fully characterized (e.g., for the synthesis of polyketides). Nonetheless, with recent advances in microalgal genomics and transcriptomic approaches, it is becoming easier to identify sequences encoding targeted enzymes, increasing the likelihood of the identification, heterologous expression, and characterization of these enzymes of interest. This review provides an overview of the state of the art in marine and freshwater microalgal enzymes with potential biotechnological applications and provides future perspectives for this field.

scholarly journals Highly efficient novel recombinant L-asparaginase with no glutaminase activity from a new halo-thermotolerant Bacillus strain

Bioimpacts ◽  
2018 ◽  
Vol 9 (1) ◽  
pp. 15-23 ◽  
Author(s):  
Azam Safary ◽  
Rezvan Moniri ◽  
Maryam Hamzeh-Mivehroud ◽  
Siavoush Dastmalchi
Keyword(s):  
Lymphoblastic Leukemia ◽  
Industrial Applications ◽  
Biochemical Properties ◽  
Substrate Affinity ◽  
Bacillus Sp ◽  
Recombinant Enzymes ◽  
E Coli ◽  
Bacillus Strain ◽  
Antineoplastic Effect ◽  
Wide Range

Introduction: The bacterial enzyme has gained more attention in therapeutic application because of the higher substrate specificity and longer half-life. L-asparaginase is an important enzyme with known antineoplastic effect against acute lymphoblastic leukemia (ALL). Methods: Novel L-asparaginase genes were identified from a locally isolated halo-thermotolerant Bacillus strain and the recombinant enzymes were overexpressed in modified E. coli strains, OrigamiTM B and BL21. In addition, the biochemical properties of the purified enzymes were characterized, and the enzyme activity was evaluated at different temperatures, pH, and substrate concentrations. Results: The concentration of pure soluble enzyme obtained from Origami strain was ~30 mg/L of bacterial culture, which indicates the significant improvement compared to L-asparaginase produced by E. coli BL21 strain. The catalytic activity assay on the identified L-asparaginases (ansA1 and ansA3 genes) from Bacillus sp. SL-1 demonstrated that only ansA1 gene codes an active and stable homologue (ASPase A1) with high substrate affinity toward L-asparagine. The Kcat and Km values for the purified ASPase A1 enzyme were 23.96s-1 and 10.66 µM, respectively. In addition, the recombinant ASPase A1 enzyme from Bacillus sp. SL-1 possessed higher specificity to L-asparagine than L-glutamine. The ASPase A1 enzyme was highly thermostable and resistant to the wide range of pH 4.5–10. Conclusion: The biochemical properties of the novel ASPase A1 derived from Bacillus sp. SL-l indicated a great potential for the identified enzyme in pharmaceutical and industrial applications.

Applicatiions and Research of Data Mining in Teaching

2011 ◽  
Vol 58-60 ◽  
pp. 2659-2663
Author(s):  
Jiang Yi Du ◽  
Yi Meng Chen
Keyword(s):  
Data Mining ◽  
Mining Technology ◽  
Network Education ◽  
Useful Knowledge ◽  
Education Data ◽  
Positive Effect ◽  
Retail Finance

Data mining technology has been widely used in the retail, finance, telecommunications and many other industries. With the promotion of education informationiation, useing the data mining technology in network education , finding useful knowledge in large education data to guide education and develop education become a necessary research. Based on the descriptionof the concept，characteristics, methods, and implement process of data mining, this paper introduces its several applications in teaching and the positive effect of teaching.

Biosynthesis of Ethyl (S)-4-Chloro-3-Hydroxybutanoate by NADH-Dependent Reductase from E. coli CCZU-Y10 Discovered by Genome Data Mining Using Mannitol as Cosubstrate

2014 ◽  
Vol 173 (8) ◽  
pp. 2042-2053 ◽  
Author(s):  
Yu-Cai He ◽  
Zhen-Xing Yang ◽  
Dan-Ping Zhang ◽  
Zhi-Cheng Tao ◽  
Chao Chen ◽  
...  
Keyword(s):  
Data Mining ◽  
E Coli ◽  
Genome Data

scholarly journals Identification and Biochemical Characterization of a Novel Protein Phosphatase 2C-Like Ser/Thr Phosphatase in Escherichia coli

2018 ◽  
Vol 200 (18) ◽  
Author(s):  
Krithika Rajagopalan ◽  
Elizabeth Nagle ◽  
Jonathan Dworkin
Keyword(s):  
Escherichia Coli ◽  
Protein Phosphatase ◽  
Biochemical Characterization ◽  
Regulatory Protein ◽  
Negative Bacterium ◽  
Gram Negative ◽  
Content Type ◽  
E Coli ◽  
Novel Protein

Regulatory protein phosphorylation is a conserved mechanism of signaling in all biological systems. Recent phosphoproteomic analyses of phylogenetically diverse bacteria, including the model Gram-negative bacteriumEscherichia coli, demonstrate that many proteins are phosphorylated on serine or threonine residues. In contrast to phosphorylation on histidine or aspartate residues, phosphorylation of serine and threonine residues is stable and requires the action of a partner Ser/Thr phosphatase to remove the modification. Although a number of Ser/Thr kinases have been reported inE. coli, no partner Ser/Thr phosphatases have been identified. Here, we biochemically characterize a novel Ser/Thr phosphatase that acts to dephosphorylate a Ser/Thr kinase that is encoded in the same operon.

REGULATION OF ACETOHYDROXY ACID SYNTHETASE IN STREPTOMYCIN-DEPENDENT ESCHERICHIA COLI K-12

1966 ◽  
Vol 44 (5) ◽  
pp. 607-612 ◽  
Author(s):  
W. J. Polglase
Keyword(s):  
Escherichia Coli ◽  
Optical Density ◽  
Density Increase ◽  
Regulatory Effect ◽  
Michaelis Constant ◽  
E Coli ◽  
K 12 ◽  
Positive Effect

The formation of acetohydroxy acid (AHA) synthetase in streptomycin-dependent Escherichia coli K-12 as a function of antibiotic (dihydrostreptomycin) concentration can be expressed by a Michaelis constant (KM = 2 × 10−4M). Valine exerts end-product repression (partial) of AHA synthetase in competition with the positive regulatory effect of dihydrostreptomycin. In dependent E. coli K-12, growth (as measured by optical density increase) correlated closely with the formation of AHA synthetase. The observed positive effect of dihydrostreptomycin on AHA synthetase formation suggests that derepression of this enzyme may be obligatory in antibiotic-dependent cells.

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