Botulinum neurotoxins are diverse proteins. They are currently represented by at least seven serotypes and more than 40 subtypes. New clostridial strains that produce novel neurotoxin variants are being identified with increasing frequency, which presents challenges when organizing the nomenclature surrounding these neurotoxins. Worldwide, researchers are faced with the possibility that toxins having identical sequences may be given different designations or novel toxins having unique sequences may be given the same designations on publication. In order to minimize these problems, an ad hoc committee consisting of over 20 researchers in the field of botulinum neurotoxin research was convened to discuss the clarification of the issues involved in botulinum neurotoxin nomenclature. This publication presents a historical overview of the issues and provides guidelines for botulinum neurotoxin subtype nomenclature in the future.

https://www.mdpi.com/2072-6651/9/1/38/pdf

Historical perspectives and guidelines for botulinum neurotoxin subtype nomenclature

We sequenced the 2 botulinum toxin gene clusters of Clostridium botulinum strain IBCA10-7060 type Bh. The sequence of bont/H differed substantially from the sequences of the 7 known bont genes for toxin types A-G. The 5' one-third terminus of bont/H that codes for the botulinum toxin light chain differed markedly from the light chain coding sequences of toxin types A-G. The 3' two-thirds terminus of bont/H that codes for the botulinum toxin heavy chain contained a novel Hn translocation domain coding sequence and a nonneutralizing type A-like Hc binding domain coding sequence. bont/H was part of an orfX toxin gene cluster that was located at a unique chromosomal site distant from those used by other botulinum toxin gene clusters. The bont/B sequence was similar to that of subtype bont/B2 and was located within its ha toxin gene cluster at the oppA/brnQ site. Our findings further establish that C. botulinum IBCA10-7060 produces novel BoNT/H.

/pdf/molecular-characterization-of-a-novel-botulinum-neurotoxin-2fazql5k3i.pdf

Molecular Characterization of a Novel Botulinum Neurotoxin Type H Gene

The ability to form botulinum neurotoxin is restricted to six phylogenetically and physiologically distinct bacteria (Clostridium botulinum Groups I-IV and some strains of C. baratii and C. butyricum). The botulinum neurotoxin is the most potent toxin known, with as little as 30-100 ng potentially fatal, and is responsible for botulism, a severe neuroparalytic disease that affects humans, animals, and birds. In order to minimize the hazards presented by the botulinum neurotoxin-forming clostridia, it is necessary to extend understanding of the biology of these bacteria. Analyses of recently available genome sequences in conjunction with studies of bacterial physiology are beginning to reveal new and exciting information on the biology of these dangerous bacteria. At the whole organism level, substantial differences between the six botulinum neurotoxin-forming clostridia have been reported. For example, the genomes of proteolytic C. botulinum (C. botulinum Group I) and non-proteolytic C. botulinum (C. botulinum Group II) are highly diverged and show neither synteny nor homology. It has also emerged that the botulinum neurotoxin-forming clostridia are not overtly pathogenic (unlike C. difficile), but saprophytic bacteria that use the neurotoxin to kill a host and create a source of nutrients. One important feature that has contributed to the success of botulinum neurotoxin-forming clostridia is their ability to form highly resistant endospores. The spores, however, also present an opportunity to control these bacteria if escape from lag phase (and hence growth) can be prevented. This is dependent on extending understanding of the biology of these processes. Differences in the genetics and physiology of spore germination in proteolytic C. botulinum and non-proteolytic C. botulinum have been identified. The biological variability in lag phase and its stages has been described for individual spores, and it has been shown that various adverse treatments extend different stages of lag phase. For example, heat treatment primarily extended germination, while incubation at a chilled temperature primarily extended outgrowth. The neurotoxin gene is present within a cluster of associated genes, and can be located on the chromosome, a plasmid or a bacteriophage. Two basic types of neurotoxin cluster have been identified. Evolution of the neurotoxin gene and cluster has occurred independently of the organism, and involved a series of recombination events but is still poorly understood. Factors affecting the regulation of neurotoxin formation also remain poorly understood, and will be the focus of much future research.

Biology and Genomic Analysis of Clostridium botulinum

Botulinum neurotoxins are known to have seven serotypes (BoNT/A-G). Here we report a new BoNT serotype, tentatively named BoNT/X, which has the lowest sequence identity with other BoNTs and is not ...

/pdf/identification-and-characterization-of-a-novel-botulinum-1hlo0jss1r.pdf

Identification and characterization of a novel botulinum neurotoxin.

Clostridium botulinum is a species of spore-forming anaerobic bacteria defined by the expression of any one or two of seven serologically distinct botulinum neurotoxins (BoNTs) designated BoNT/A-G. This Gram-positive bacterium was first identified in 1897 and since then the paralyzing and lethal effects of its toxin have resulted in the recognition of different forms of the intoxication known as food-borne, infant, or wound botulism. Early microbiological and biochemical characterization of C. botulinum isolates revealed that the bacteria within the species had different characteristics and expressed different toxin types. To organize the variable bacterial traits within the species, Group I–IV designations were created. Interestingly, it was observed that isolates within different Groups could express the same toxin type and conversely a single Group could express different toxin types. This discordant phylogeny between the toxin and the host bacteria indicated that horizontal gene transfer of the toxin was responsible for the variation observed within the species. The recent availability of multiple C. botulinum genomic sequences has offered the ability to bioinformatically analyze the locations of the bont genes, the composition of their toxin gene clusters, and the genes flanking these regions to understand their variation. Comparison of the genomic sequences representing multiple serotypes indicates that the bont genes are not in random locations. Instead the analyses revealed specific regions where the toxin genes occur within the genomes representing serotype A, B, C, E, and F C. botulinum strains and C. butyricum type E strains. The genomic analyses have provided evidence of horizontal gene transfer, site-specific insertion, and recombination events. These events have contributed to the variation observed among the neurotoxins, the toxin gene clusters and the bacteria that contain them, and has supported the historical microbiological, and biochemical characterization of the Group classification within the species.

Genetic Diversity Within Clostridium botulinum Serotypes, Botulinum Neurotoxin Gene Clusters and Toxin Subtypes

Plasmid-encoded neurotoxin genes in clostridium botulinum serotype a subtypes

The present invention relates to compositions and methods for testing agents (e.g., Clostridium botulinum neurotoxin (BoNT) detection and analysis). In particular, the present invention relates to the use of Human induced pluripotent stem (hiPS) derived cells for agent detection and analysis.

Compositions and methods for toxigenicity testing

The present invention provides a universal delivery platform of functional, heterologous compounds to specific cells using toxins modified to include a heterologous compound. In one embodiment, the toxin is an AB5 toxin. In one embodiment, the AB5 toxin is a heat-labile enterotoxin from E. coli (LT), including LTI, LTII, LTIIa, LTIIb, LTIIc and other recombinant forms of LT. Methods of use are also provided.

Universal platform for targeting therapies to treat neurological diseases

Covid-19 vaccine

The present disclosure relates to neurotoxins and uses thereof. In particular, provided herein are botulinum neurotoxins with altered properties and uses thereof (e.g., research, screening, and therapeutic uses).

Neurotoxins and uses thereof

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