Stanislav Pepeliaev

Stanislav Pepeliaev
University of Oxford | OX · Jenner Institute

Doctor of Philosophy

About

17
Publications
1,372
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105
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Introduction
Upstream development of neurotoxins recombinant production. Strong interdisciplinary skills including metabolic and strain engineering, protein expression and purification, upstream and downstream processes development.

Publications

Publications (17)
Article
Human enterokinase (enteropeptidase, rhEP), a serine protease expressed in the proximal part of the small intestine, converts the inactive form of trypsinogen to active trypsin by endoproteolytic cleavage. The high specificity of the target site makes enterokinase an ideal tool for cleaving fusion proteins at defined cleavage sites. The mature acti...
Patent
Full-text available
The invention relates to a method of determination of hyaluronic acid in complex samples by cleavage with SpnHyl enzyme followed by the reaction with 3-methyl-2-benzothiazolinone hydrazone (MBTH) to form a coloured product analysed by spectrophotometry. In case of a high content of glycosaminoglycans, these can be separated by precipitation before...
Patent
Full-text available
There is provided a method of controlling operation of a fed batch process in a bioreactor vessel, comprising transitioning from a batch phase to a production phase in dependence on a relationship between an oxygen supply parameter O and a dissolved oxygen value DO.
Article
The physicochemical properties and biological functions of hyaluronan (HA) are closely related to its molecular weight (MW) and molecular weight distribution (MWD). Therefore, it is crucially important to provide a reliable characterization of these parameters for proper use of HA and its degradation products in both chemical and clinical fields. I...
Conference Paper
Full-text available
Demanding safety standards needed to protect personnel during botulinum neurotoxin (BoNT)-based pharmaceutical production can hinder development of these products, requiring extensive safety measures. However, speed-to-market is crucial to provide patients with new treatments without delay and reduce costs. We sought to accelerate BoNT drug develop...
Conference Paper
Hexosaminidase from Streptomyces plicatus is an enzyme that catalyzes hydrolytic cleavage of N-acetylglucosamine from oligosaccharides. Its activity on substrates such as chitin oligosaccharides and glycans from glycoproteins are known. Here we describe activity of this enzyme towards hyaluronic acid oligosaccharides. Hyaluronic acid is a linear po...
Conference Paper
The natural competence is the genetically specified ability of some bacteria to alter their genome by taking up foreign extracellular DNAfrom its environment. Initially it was found in Streptococcus pneumonia, later it was described for many species of Streptococci genus and for other groups of bacteria. The mechanism of natural competence is well...
Article
Hyaluronic acid (HA) is a component of the extracellular matrix with important potential for biotechnological and pharmaceutical applications. However, the immunological effects of HA are strongly molecular weight-dependent; therefore, production of HA for therapeutic purposes requires control over the molecular weight of the product, as well as mi...
Conference Paper
Streptococcus equi subsp. zooepidemicus is a gram-positive bacterium of the Lancefield group C streptococci. It is known to produce a hyaluronic acid capsule that resists the host’s immune defence. Hyaluronic acid is a linear polysaccharide of high molecular weight. It occurs naturally in the human body, where it is applied as a connective tissue o...
Presentation
Hyaluronan (HA) and its derivatives are clinically used to treat diseases or they are employed in cosmetic surgery procedures. To enhance, modulate or control the therapeutic action, the chemical modifications of HA have been previously described in literature. Peptides prepared and presented on hyaluronan scaffolds added significant and valuable p...
Article
Full-text available
Hyaluronic acid (HA), a high molecular glycosaminoglycan, is a chief component of the extracellular matrix in vertebrates. It fulfils several important biological functions, among which structural and regulatory are the most important. Hyaluronidases (HAases) are enzymes that degrade predominantly HA. HAases from vertebrate tissues, pathogenic bact...
Article
Hyaluronidases (HAases) from yeasts were characterized for the first time. The study elucidated that hyaluronate 4-glycanohydrolase and hyaluronan (HA) lyase can be produced by yeasts. Six yeasts producing HAases were found through express screening of activities. The extracellular HAases from two of the yeast isolates, Pseudozyma aphidis and Crypt...
Article
This study addresses the influence of upstream region sequence on the strength of has operon promoter in highly encapsulated S. equi subsp. zooepidemicus (SEZ). For this purpose, seven different strains were constructed. Each strain carries a point mutation in one of the following positions upstream of the has promoter: −43, −44, −49, and −50 bp. T...
Article
Full-text available
The light chain of human enteropeptidase (enterokinase) is one of the most frequently used proteases because of its high specificity and activity. It cleaves proteins after the sequence (Asp)4Lys and leaves N-terminus of the downstream fused protein intact, it is stable in a broad range of pH and denaturing agent concentration. This work presents s...
Article
The human antimicrobial peptide LL-37 is a cationic peptide with antimicrobial activity against both Gram-positive and Gram-negative microorganisms. This work describes the development of an expression system based on Escherichia coli capable of high production of the recombinant LL-37. The fusion protein Trx-LL-37 was expressed under control of T7...

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