An N-Terminal Protein Degradation Tag Enables Robust Selection of Highly Active Enzymes

Biochemistry ◽  
2011 ◽  
Vol 50 (40) ◽  
pp. 8594-8602 ◽  
Author(s):  
Maren Butz ◽  
Martin Neuenschwander ◽  
Peter Kast ◽  
Donald Hilvert
Keyword(s):  
Protein Degradation ◽  
Terminal Protein ◽  
Highly Active ◽  
Selection Of

THE DETERMINATION OF TERMINAL PROTEIN-BOUND FUCOSE

1965 ◽  
Vol 43 (11) ◽  
pp. 1807-1811 ◽  
Author(s):  
G. Gyorky ◽  
J. C. Houck
Keyword(s):  
Protein Degradation ◽  
Spectrophotometric Determination ◽  
Trichloroacetic Acid ◽  
Degradation Products ◽  
Dilute Acid ◽  
Terminal Protein ◽  
Residual Protein ◽  
Color Yield ◽  
Fucose Content

The spectrophotometric determination of protein-bound fucose is badly compromised by spurious chromagens developed from protein degradation products. To minimize the contribution of these spurious products to the color yield of fucose, the glycoprotein was partially hydrolyzed in dilute acid, thus releasing the terminal carbohydrate from the protein moieties, and the residual protein was removed with trichloroacetic acid. That the fucose content of this supernatant was real was confirmed by paper chromatography and spectral studies.The spurious chromagens were shown to result from the interaction of protein degradation products and galactose.

Does N‐Terminal Protein Acetylation Lead to Protein Degradation?

BioEssays ◽  
2019 ◽  
Vol 41 (11) ◽  
pp. 1800167 ◽  
Author(s):  
Mohamed A. Eldeeb ◽  
Richard P. Fahlman ◽  
Mohamed A. Ragheb ◽  
Mansoore Esmaili
Keyword(s):  
Protein Degradation ◽  
Protein Acetylation ◽  
Terminal Protein

scholarly journals Efficient CRISPR/Cas9-mediated biallelic gene disruption and site-specific knockin after rapid selection of highly active sgRNAs in pigs

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Xianlong Wang ◽  
Jinwei Zhou ◽  
Chunwei Cao ◽  
Jiaojiao Huang ◽  
Tang Hai ◽  
...  
Keyword(s):  
Gene Disruption ◽  
Site Specific ◽  
Highly Active ◽  
Selection Of

scholarly journals Biologically Active Aldehydes In Extracts Of Lactarius Pergamenus (Fr.) Fr Fresh Fruiting Bodies

2020 ◽  
Vol 15 (3) ◽  
pp. 125-131
Author(s):  
V.O. Antonyuk ◽  
L.V. Panchak ◽  
M.V. Tsivinska ◽  
R.S. Stoika
Keyword(s):  
Thin Layer ◽  
Aromatic Amines ◽  
Methylene Chloride ◽  
Biologically Active ◽  
Fruiting Bodies ◽  
Highly Active ◽  
Vegetable Kingdom ◽  
Methylene Chloride Extract ◽  
Colored Compound ◽  
Selection Of

The biologically active aldehydes in extracts of fungi of the genus Lactarius were identified. It’s established that these substances are unstable, they are found in fresh and frozen fungi, but are absent in dried mushrooms and interact with 1,4-phenylenediamine to form a colored compound. Methylene chloride is the best extragent for these substances. TLC on silufol plates showed that there were several substances in Lactarius pergamenus fruiting bodies and they had varying degrees of stability. For selection of these substances, methylene chloride extract was separated on a column of silica gel. Fraction, which gave the most expressive reaction with 1,4-phenylenediamine on thin-layer chromatograms were analyzed by GC-MS both in the absence and in the presence of 1,4-phenylenediamine. As a result, it was found that 1,4-phenylenediamine or other aromatic amines interacted with highly active aldehydes, that were present in fruiting bodies. Among them 2,2-dimethylocta- 3,4-dienal was the most stable and was present in the biggest quantity. This substance very rarely occurs in the vegetable kingdom and in fungi extracts of Lactarius genus wasn’t previously described. Its possible function in fungi is prevention of damage by parasites and eating by animals.

scholarly journals РАЗРАБОТКА ВЫСОКОАКТИВНОГО ШТАММА-ПРОДУЦЕНТА ВИРДЖИНИАМИЦИНА И ПОВЫШЕНИЕ ЕГО ПРОДУКТИВНОСТИ С ПОМОЩЬЮ СИНТЕТИЧЕСКИХ АБСОРБИРУЮЩИХ СМОЛ

2016 ◽  
Vol 6 (3) ◽  
pp. 195-208 ◽  
Author(s):  
V. A. Savushkin ◽  
V. V. Dzhavakhiya ◽  
E. V. Glagoleva ◽  
V. V. Savel'eva ◽  
E. D. Popova ◽  
...  
Keyword(s):  
Fermentation Medium ◽  
Culture Broth ◽  
Medium Composition ◽  
Basic Medium ◽  
High Productivity ◽  
Highly Active ◽  
Different Types ◽  
Total Productivity ◽  
Industrial Use ◽  
Selection Of

<p>Virginiamycin, an antibiotic produced by some <em>Streptomyces</em> species, is widely used in veterinary and bioethanol production. It represents a natural mix of two different macrocyclic components, among which M1 and S1 factors are the main acting components. M1 and S1 act synergistically when present in the optimum ratio of 60-75% of M1 and 25-40% of S1. Due to a large number of genes involved into the virginiamycin biosynthesis, the development of overproducing strains able to synthesize M1 and S1 at a synergistic ratio with the total productivity exceeding 3-4 g/L still remains a relevant problem. Using a multi-step random UV mutagenesis of the <em>Streptomyces</em> sp. strain DSM40559, a highly active strain S 15-30 was obtained, which virginiamycin titer on a basic medium significantly increased that of the parental strain (2.6 and 0.35 g/L, respectively), and the M1:S1 ratio remained synergistic (72:28). Various sources of carbon, nitrogen, and macroelements were evaluated for medium improvement, and several different types of synthetic macroporous resins were tested to provide the highest virginiamycin titer in culture broth of the developed strain. The resulting improved fermentation medium supplemented with 20 g/L of Amberlite XAD-16 resin increased strain productivity up to 5.03 ± 0.12 g/L with the simultaneous maintenance of the M1:S1 ratio within the synergistic range (72:28) and highly selective level of adsorption of the antibiotic from culture broth (92-95%). The variability of the M1:S1 ratio in the total antibiotic titer depending on various medium composition and resin type was first demonstrated. The obtained strain is promising for the industrial use due to its high productivity and the optimal M1:S1 ratio. Results of the study will be used for the further selection of overproducing strains and the scaling up of the virginiamycin production. The obtained data can be interesting for other researchers working in the field of production of macrolide antibiotics.</p>

scholarly journals Effective polymer concrete on waste concrete production

2019 ◽  
Vol 97 ◽  
pp. 02032
Author(s):  
Evgeny Pyataev ◽  
Alexey Zhukov ◽  
Kirill Vako ◽  
Marina Burtseva ◽  
Elizaveta Mednikova ◽  
...  
Keyword(s):  
Average Density ◽  
Polymer Concrete ◽  
Polymer Additive ◽  
Highly Active ◽  
Active Experiment ◽  
High Durability ◽  
Concrete Production ◽  
Building Systems ◽  
Active Waste ◽  
Selection Of

Finish materials in facade systems are experiencing significant stress associated with atmospheric and mechanical influences. The use of tiles with high durability and the ability to relax deformations will allow to obtain efficient cladding for building systems. The article presents the results of the selection of compositions and technologies of a composite material based on finely ground highly active waste produced by portland cement, mineral binder, additives based on polycarboxylic ether MC-6955, and polymer additive MC-Adhesive. MC-Adhesive is a polymer additive used for: a significant increase in flexural strength; reduce the elastic modulus; increase water resistance; increase the concrete mix cohesion; making coatings with high requirements for abrasion, low dusting and high resistance to aggressive substances.The article presents the results of research methods of selection of compositions of composite polymer concrete. The strength characteristics of polymer concrete with an average density of 2370-2450 kg/m3 are most dependent on the consumption of fine waste and polymer additives. The nomogram obtained as a result of an active experiment makes it possible to evaluate the dependence of the strength on these factors and select their optimal ratio.

A Novel, Universally Active C-terminal Protein Degradation Signal Generated by Alternative Splicing

2021 ◽  
Vol 433 (8) ◽  
pp. 166890
Author(s):  
Stephen A. Peter ◽  
Jessica S. Isaac ◽  
Franz Narberhaus ◽  
Julia E. Weigand
Keyword(s):  
Alternative Splicing ◽  
Protein Degradation ◽  
Terminal Protein

Lanthanide-dependent RNA-cleaving DNAzymes as metal biosensors

2015 ◽  
Vol 93 (3) ◽  
pp. 273-278 ◽  
Author(s):  
Juewen Liu
Keyword(s):  
Sensor Array ◽  
Activity Patterns ◽  
Chemical Properties ◽  
Toxic Heavy Metals ◽  
Future Directions ◽  
Highly Active ◽  
Lanthanide Series ◽  
Rational Ligand Design ◽  
Heavy Lanthanides ◽  
Selection Of

Lanthanides represent a group of very important but challenging analytes for biosensor development. These 15 elements are very similar in their chemical properties. So far, limited success has been realized using the rational ligand design approach. My laboratory has successfully accomplished the task of carrying out combinatorial selection to isolate lanthanide-dependent RNA-cleaving DNAzymes. We report two new DNAzymes, each discovered in a different selection condition and both are highly specific to lanthanides. When both DNAzymes are used together, it is possible to identify the last few heavy lanthanides. Upon introducing a phosphorothioate modification, one of the abovementioned DNAzymes becomes highly active with many toxic heavy metals. With the selection of more DNAzymes with different activity patterns cross the lanthanide series, a sensor array might be produced for identifying each ion. This article is a minireview of the current developments on this topic and some of the historical aspects. It reflects the main content of the Fred Beamish Award presentation delivered at the 2014 Canadian Society for Chemistry Conference in Vancouver. Future directions in this area are also discussed.

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