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Cloning and characterization of sialidases with 2-6' and 2-3' sialyl lactose specificity from Pasteurella multocida.

ABSTRACT: Pasteurella multocida is a mucosal pathogen that colonizes the respiratory system of susceptible hosts. Most isolates of P. multocida produce sialidase activity, which may contribute to colonization of the respiratory tract or the production of lesions in an active infection. We have cloned and sequenced a sialidase gene, nanH, from a fowl cholera isolate of P. multocida. Sequence analysis of NanH revealed that it exhibited significant amino acid sequence homology with many microbial sialidases. Insertional inactivation of nanH resulted in a mutant strain that was not deficient in sialidase production. However, this mutant exhibited reduced enzyme activity and growth rate on 2-3' sialyl lactose compared to the wild type. Subsequently, we demonstrated the presence of two sialidases by cloning another sialidase gene that differed from nanH in DNA sequence and substrate specificity. NanB demonstrated activity on both 2-3' and 2-6' sialyl lactose, while NanH demonstrated activity only on 2-3' sialyl lactose. Neither enzyme liberated sialic acid from colominic acid (2-8' sialyl lactose). Recombinant E. coli containing the sialidase genes were able to utilize several sialoconjugants when they were provided as sole carbon sources in minimal medium. These data suggest that sialidases have a nutritional function and may contribute to the ability of P. multocida to colonize and persist on vertebrate mucosal surfaces.

SUBMITTER: Mizan S

PROVIDER: S-EPMC94810 | biostudies-literature | 2000 Dec

REPOSITORIES: biostudies-literature

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Cloning and characterization of sialidases with 2-6' and 2-3' sialyl lactose specificity from Pasteurella multocida.

Mizan S S Henk A A Stallings A A Maier M M Lee M D MD

Journal of bacteriology 20001201 24

Pasteurella multocida is a mucosal pathogen that colonizes the respiratory system of susceptible hosts. Most isolates of P. multocida produce sialidase activity, which may contribute to colonization of the respiratory tract or the production of lesions in an active infection. We have cloned and sequenced a sialidase gene, nanH, from a fowl cholera isolate of P. multocida. Sequence analysis of NanH revealed that it exhibited significant amino acid sequence homology with many microbial sialidases. ...[more]

PMID: 11092845

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Project description:BackgroundThis study provides biochemical and molecular genetic characteristics of P. multocida isolated from dead saigas in 1988, 2010-2015 on the territory of the Republic of Kazakhstan.ResultsBacteriological samples taken from carcasses of saiga antelope during mortality events recorded in West Kazakhstan in both 2010 and 2011 and in Kostanay in 2012 and 2015 confirmed the presence of P. multocida, according to morphological and biochemical characterisation. Only in the event of 2015 was the agent proven to be the causative agent of the disease observed, haemorrhagic septicaemia. In the other mortality events it is not certain if the organism was a primary aetiology or an incidental finding as confirmatory pathological investigation was not undertaken. The implemented phylogenetic analysis of ribosomal RNA 16S gene allowed us to identify Pasteurella strains isolated in 2010-2015 as P. multocida subspecies multocida. Capsular typing by PCR showed that the studied strains isolated from dead saiga in 2010, 2011, 2012 and 2015 belonged to serotype B. MLST analysis showed that these strains of P. multocida are of the capsule type B and form one clonal grouping with isolates ST64, ST44, ST45, ST46, ST44, ST47 which isolated from cases of hemorrhagic septicemia of animals in Hungary, Burma, Sri Lanka, Pakistan and Spain. Sixteen virulence genes of the five strains of P. multocida, isolated from saigas were studied using multiplex PCR. ptfA, ompA, ompH, oma87, plpB, fimA, hsf-2, pfhA, exbB, tonB, hgbA, fur, nanB, nanH and pmHAS genes were detected in all strains. The toxA gene was not identified in the studied strains. The phylogenies of these isolates is compared across saiga populations and years and the 2015 isolate was compared to that of an isolate from a disease outbreak in 1988 and the findings suggest that these isolated bacteria are stable commensals, opportunistically pathogenic, being phylogenetically uniform with very little genetic variation notable over the last 4 decades.ConclusionIsolation, phenotypic and genetic characterization of the P. multocida isolates inform understanding of the epidemiology of infection in saigas and predict virulent potential of these opportunistic bacteria.

Dataset Information

Cloning and characterization of sialidases with 2-6' and 2-3' sialyl lactose specificity from Pasteurella multocida.

Publications

Cloning and characterization of sialidases with 2-6' and 2-3' sialyl lactose specificity from Pasteurella multocida.

Similar Datasets

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