A5091878500- Bacteriophages and microbial interactions
- CRISPR and Genetic Engineering
- Genomics and Phylogenetic Studies
- Bacterial Genetics and Biotechnology
- RNA and protein synthesis mechanisms
- Advanced biosensing and bioanalysis techniques
- Biochemical and Molecular Research
- Alkaline Phosphatase Research Studies
- Protein purification and stability
- DNA and Nucleic Acid Chemistry
- RNA modifications and cancer
- RNA Interference and Gene Delivery
- Probiotics and Fermented Foods
- Biotin and Related Studies
- DNA Repair Mechanisms
- Microbial Inactivation Methods
- Epigenetics and DNA Methylation
- Metalloenzymes and iron-sulfur proteins
- Biofuel production and bioconversion
- Plant-Microbe Interactions and Immunity
- Electrocatalysts for Energy Conversion
- Enzyme Structure and Function
- Microbial Community Ecology and Physiology
- Fungal and yeast genetics research
- Photochemistry and Electron Transfer Studies
University of Gdańsk
2014-2024
Abstract Background One of the leading current trends in technology is miniaturization devices to microscale and nanoscale. The highly advanced approaches are based on biological systems, subjected bioengineering using chemical, enzymatic recombinant methods. Here we have utilised affinity towards cellulose binding domain (CBD) fused with proteins. Results focused fusions ‘artificial’, concatemeric proteins preprogrammed functions, constructed DNA FACE™ technology. Such CBD can be...
An industrial approach to protein production demands maximization of cloned gene expression, balanced with the recombinant host's viability. Expression toxic genes from thermophiles poses particular difficulties due high GC content, mRNA secondary structures, rare codon usage and impairing coding plasmid replication.TaqII belongs a family bifunctional enzymes, which are fusion restriction endonuclease (REase) methyltransferase (MTase) activities in single polypeptide. The contains...
Bacteriophage TP-84 is a well-characterized bacteriophage of historical interest. It member the Siphoviridae, and infects number thermophilic Geobacillus (Bacillus) stearothermophilus strains. Its' 47.7-kbp double-stranded DNA genome revealed presence 81 coding sequences (CDSs) for polypeptides 4 kDa or larger. Interestingly, all CDSs are oriented in same direction, pointing to dominant transcription direction one strand. Based on homology search, hypothetical function could be assigned 31...
Abstract Background Restriction-modification systems are a diverse class of enzymes. They classified into four major types: I, II, III and IV. We have previously proposed the existence Thermus sp. enzyme family, which belongs to type II restriction endonucleases (REases), however, it features also some characteristics types I III. Members include related thermophilic endonucleases: TspGWI, TaqII, TspDTI, Tth111II. Results Here we describe cloning, mutagenesis analysis prototype TspGWI that...
Abstract Background We previously defined a family of restriction endonucleases (REases) from Thermus sp., which share common biochemical and biophysical features, such as the fusion both nuclease methyltransferase (MTase) activities in single polypeptide, cleavage at distance recognition site, large molecular size, modulation activity by S-adenosylmethionine (SAM), incomplete substrate DNA. Members include related thermophilic REases with five distinct specificities: TspGWI, TaqII,...
Obtaining thermostable enzymes (thermozymes) is an important aspect of biotechnology. As thermophiles have adapted their genomes to high temperatures, cloned genes' expression in mesophiles problematic. This mainly due GC content, which leads the formation unfavorable secondary mRNA structures and codon usage Escherichia coli (E. coli). RM.TthHB27I a member family bifunctional thermozymes, containing restriction endonuclease (REase) methyltransferase (MTase) single polypeptide. Thermus...
The type IIS/IIC restriction endonuclease TspGWI recognizes the sequence 5′-ACGGA-3′, cleaving DNA 11/9 nucleotides downstream. Here we show that sinefungin, a cofactor analog of S-adenosyl methionine, induces unique relaxation in recognition specificity. In presence and cleaves at least 12 degenerate variants original vary by single base pair changes from 5-bp site with only degeneracy per variant appearing to be allowed. addition, sinefungin was found have stimulatory effect on cleavage...
5-Bromouracil (BrU) is photoreactive toward near UVB photons and can be introduced into genomic DNA during its biosynthesis in cells. However, PCR seems to a simpler approach, which used obtain labeled similar that synthesized within the cell. In current work, has been employed optimized order substitute all thymines (besides those present starters) with BrU dsDNA fragment of 80 base pairs (bp) length. The modified oligonucleotide was irradiated 300 nm buffered aqueous solution (pH = 7)...
In spite of the fact that recombinant enzymes are preferably biotechnologically obtained using clones, purification proteins from native microorganisms, including those encoded by bacteriophages, continues. The bacteriophage protein isolation is often troubled large volumes infected bacterial cell lysates needed to be processed, which highly undesired in scaled-up industrial processing. A well-known ammonium sulphate fractionation a method choice during protein. However, this time-consuming...
Abstract Background The TaqII enzyme is a member of the Thermus sp. family that we propounded previously within Type IIS restriction endonucleases, containing related thermophilic bifunctional endonucleases-methyltransferases from various sp.: TaqII, Tth111II, TthHB27I, TspGWI, TspDTI and TsoI. These enzymes show significant nucleotide amino acid sequence similarities, rare phenomenon among along with similarities in biochemical properties, molecular size, DNA recognition sequences cleavage...
In continuing our research into the new family of bifunctional restriction endonucleases (REases), we describe cloning tsoIRM gene. Currently, includes six thermostable enzymes: TaqII, Tth111II, TthHB27I, TspGWI, TspDTI, TsoI, isolated from various Thermus sp. and two thermolabile RpaI CchII, mesophilic bacteria Rhodopseudomonas palustris Chlorobium chlorochromatii, respectively. The enzymes have several properties in common. They are large proteins (molecular size app. 120 kDa), coded by...
The TP-84 bacteriophage, which infects Geobacillus stearothermophilus strain 10 (G. stearothermophilus), has a genome size of 47.7 kilobase pairs (kbps) and contains 81 predicted protein-coding ORFs. One these, TP84_26 encodes putative tail fiber protein possessing capsule depolymerase activity. In this study, we cloned the gene into high-expression Escherichia coli (E. coli) system, modified its N-terminus with His-tag, expressed both wild type His-tagged variant, purified recombinant...
Bacteriophages associated with thermophiles are gaining increased attention due to their pivotal roles in various biogeochemical and ecological processes, as well applications biotechnology bionanotechnology. Although thermophages not suitable for controlling bacterial infections humans or animals, individual components, such enzymes capsid proteins, can be employed molecular biology significantly contribute the enhancement of human animal health. Despite significance, still remain...
Interpretation :The main objective of the study was to characterize a mixture bacterial species, found in commercial probiotic preparation and originally designed for cleaning, biodegradation wastewater treatment.Lyophilized environmental strains microbiologically characterized determine growth temperature range, pH resistance boiling survivability.Gram staining Wirtz's spores were performed microscopic estimation cell morphology sporulation.The MALDI-TOF mass spectrometry method used...
A novel prototype class-IIS restriction endonuclease, Tsp GWI, was isolated from the thermophilic bacterium Thermus sp. GW. The recognition sequence and cleavage positions have been established: GWI recognizes non-palindromic 5-bp 5′‐ACGGA-3′ cleaves DNA 11 9 nt downstream in top bottom strand, respectively. In addition, an accompanying GWII, isoschizomer of Pst I, found GW cells.
Abstract Background Genomics and metagenomics are currently leading research areas, with DNA sequences accumulating at an exponential rate. Although enormous advances in sequencing technologies taking place, progress is frequently limited by factors such as genomic contig assembly generation of representative libraries. A number fragmentation methods, hydrodynamic sharing, sonication or DNase I fragmentation, have various drawbacks, including damage, poor control, irreproducibility...
Abstract Background The biotechnology production of enzymes is often troubled by the toxicity recombinant products cloned and expressed genes, which interferes with hosts’ metabolism. Various approaches have been taken to overcome these limitations, exemplified tight control genes or secretion proteins. An industrial approach protein demands maximum possible yields biosynthesized proteins, balanced host’s viability. Bacterial alkaline phosphatase (BAP) from Escherichia coli ( E. ) a key...
De novo designed bioactive molecules, such as DNA, RNA and peptides, are utilized in increasingly diverse scientific, industrial biomedical applications. Concatemerization of peptides may improve their stability, bioactivity allow for gradual release the molecule at intended destination. In this context, we developed a new method enabling formation DNA concatemers production artificial, repetitive genes, encoding concatemeric RNAs proteins any nucleotide amino-acid sequence. The technology...
Two restriction-modification systems have been previously discovered in Thermus aquaticus YT-1. TaqI is a 263-amino acid (aa) Type IIP restriction enzyme that recognizes and cleaves within the symmetric sequence 5'-TCGA-3'. TaqII, contrast, 1105-aa IIC restriction-and-modification enzyme, one of family homologs. TaqII was originally reported to recognize two different asymmetric sequences: 5'-GACCGA-3' 5'-CACCCA-3'. We cloned taqIIRM gene, purified recombinant protein from Escherichia coli,...