Specialists in the art will also understand that having common way to create mutations in any of the ORF, designated SEQ ID NOS: 1, these methods can be used to alter levels of expression of the same ORF. Examples of how this can be achieved include, but are not limited to, integration of multiple copies of the specified ORF in any place in the genome of Microbispora sp. MOUTH 5024, change promoters controlling the expression of these ORFS, destruction of antisense RNA or transcription terminators, affecting their expression.
SUBSTANCE: invention describes nucleotide sequence (dwg.2), coding immunogenic polypeptide LcrV(G113), serving the base for construction of recombinant plasmid DNA pETV-I-3455, with the size 6538 bp, which codes immunogenic polypeptide LcrV(G113). Plasmid consists of plasmid pBR322 replicon, β-lactamase gene, determining resistance to ampicillin, T7-promoter, 1ac-operator, f1-replicon and DNA fragment, flanked by the sites for restrictases Ndel and Hindlll, coding synthesis of protein LcrV(G113), which starts from initiating codon ATG. Described is recombinant strain of bacteria E. coli BL21 (DE3)/pETV-I-3455 - producer of immunogenic polypeptide LcrV(G113) with amino acid sequence, represented on dwg.3, where tryptophan in position 113 (W113) is substituted with glycin. Described is method of obtaining said polypeptide by cultivation of strain E. coli BL21(DE3)/pETV-I-3455. Cells are destroyed in buffer solution by ultrasound and polypeptide is isolated successively by gel-permeation chromatography with application of carrier TSK HW-40, anion-exchanging and hydrophobic chromatography.
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SUBSTANCE: construction of recombinant plasmid DNAs containing a structured gene of human interferon alpha-2b under control of a sensitive promoter enables producing Escherichia coli strain to enable synthesis in the form of an insoluble precursor protein of mature methionine-free human interferon alpha-2b. In the precursor protein, a sequence of mature methionine-free human interferon alpha-2b is fused with a sequence of SUMO (SMT3) Saccharomyces cerevisiae yeast protein. What is produced is the Escherichia coli strain providing biosynthesis of ULP275 proteinase for enzymatic processing of the precursor protein of methionine-free interferon. What is presented is a method for producing methionine-free interferon which provides carrying out the coupled processes of renaturation, disulphide link formation and enzymatic segregation of methionine-free human interferon alpha-2b from the precursor protein under ULP275 proteinase.
We review the regulation of antibiotic biosynthesis in ..
Antibiotic 107891 demonstrates antibacterial activity against gram-positive bacteria, including methicillin - and vancomycin-resistant strains, but shows limited activity against gram-negative bacteria (for example, some Moraxella, Neisseria, and Haemophilus spp.). 107891 was isolated after fermentation Microbispora sp. PTA-5024 (WO 2005/04628 A1). It consists of a complex of closely related factors A1 and A2, the structure of which can be converted into the peptide skeleton with a length of 24 amino acids containing lanthionine and metallothionein as components. In addition, the molecule contains an atom of chlorine and one or two of rest-HE. The structure of the components of 107891 complex represented by formula 1 in figure 3, where R is a [IT] with the actor A1(R=OH), factor2(R=-(OH)2). 107891, apparently, combines elements of lantibiotics type a and b: rings a and b in 107891 closely associated with equivalent rings in compounds of the type A; however, as in antibiotico type, 107891 is rather globular, it does not contain a flexible C-terminal sequences and deprived of charged amino acid residues. Consequently, it is impossible to predict encodes whether the cluster lan, involved in education 107891, the only LanM enzyme or individual proteins LanB and LanC. In addition, there is no prerequisite for chlorinated lantibiotics thus, the genes responsible for this post-translational modification, cannot be predicted according to the available data.
Molecular Cloning and Characterization of a 2-Deoxystreptamine ..
Actually only unusual amino acids in antibiotico contribute to their biological activity, as well as increase their structural stability. The enzymes involved in the biosynthesis of lantibiotics represent a high potential for the design of peptides by introducing unusual amino acids in the desired peptides.
BioMed Research International - Hindawi Publishing …
The constant increase of pathogenic bacteria resistant to existing antibiotics is a global health risk, and thus, there is an urgent need to discover and develop new compounds that are active against resistant bacteria. The latter leads to renewed interest in natural products, which in the past predstavlyalosb a rich source of antibiotics, such as penicillins, macrolides and glycopeptides. Attractive candidates are also antimicrobial peptides and the peptides, referred to as lantibiotic, i.e. lanthionine-containing antibiotics. Lantibiotic form a special group among the antimicrobial peptides and differ in several characteristics, such as characteristics of the primary and spatial patterns, unique biosynthesis pathway and reaction modification of peptides and strong antibacterial activity. They are a group of peptide antimicrobial compounds secreted by gram-positive bacteria, and primarily act on gram-positive bacteria. Lantibiotic are synthesized on ribosomes in the form of reprobation that excision of modified prior to their biologically active forms. Propeptide consists of N-terminal leader sequence, which is not subjected to any post-translational modification and is cleaved during or after secretion from the cell, and the C-terminal region (propeptide), which is modified excision. Lantibiotic are produced by different bacteria: a common property of these compounds is the presence of one or more than one residue of lanthionine, which consists of two alanine residues, covalently crosslinked by thioester bond. jefery bridge is formed then when the residue is cysteine reacts with dehydroalanine or dehydroretinol group education respectively mentionedabove or metallothionine balance. The remains of dehydrolinalool, in turn, are formed by dehydration, respectively, serine and threonine. On the basis of structural and functional properties of lantibiotic are usually divided into two groups, type a and type B. Lantibiotic type And are extra long, cationic peptides with length varying from 20 to 34 amino acid residues: lowlands, subtilin, epidermin and RER are members of this group. The type represents a globular peptides with the General negative charge: examples of this group of lantibiotics are mersacidin, cinnarizin, lacticin 481 and actigard. These structural differences affect the mechanism of action. Connection type And carry out their antimicrobial activity by blocking the biosynthesis of the cell wall and the formation of pores in cell membranes by a mechanism, which can contribute or not to contribute to the pre-consolidation at the cellular lipid target II. Lantibiotic type also show their antimicrobial activity by inhibiting the biosynthesis of composition, but these compounds do not form pores after binding to lipid II.