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B J Stevens

Publications and source records attributed to B J Stevens.

At least 19 recordsLinked to original sources

Mechanism of action of cyclic beta-1,2-glucan synthetase from Agrobacterium tumefaciens: competition between cyclization and elongation reactions.

We have examined some aspects of the mechanism of cyclic beta-1,2-glucan synthetase from Agrobacterium tumefaciens (235-kDa protein, gene product of the chvB region). The enzyme produces cyclic beta-1,2-glucans containing 17 to 23 glucose residues from UDP-glucose. In the presence of added cyclic beta-1,2-glucans (> 0.5 mg/ml) (containing 17 to 23 glucose residues), the enzyme instead synthesizes larger cyclic beta-1,2-glucans containing 24 to 30 glucose residues. This is achieved by de novo synthesis and not by disproportion reactions with the added product. This is interpreted as inhibition of the specific cyclization reaction for the synthesis of cyclic beta-1,2-glucans containing 17 to 23 glucose residues but with no concomitant effect on the elongation (polymerization) reaction. Temperature and detergents both affect the distribution of sizes of cyclic beta-1,2-glucans, but glucans containing 24 to 30 glucose residues are not produced. We suggest that the size distribution of cyclic beta-1,2-glucan products depends on competing elongation and cyclization reactions.

Agrobacterium tumefaciens

Use of oligonucleotide probes to identify members of two-component regulatory systems in Xanthomonas campestris pathovar campestris.

Two-component regulatory systems comprising a sensor and a regulator protein, both with highly conserved amino acid domains, and commonly genetically linked, have been described in a range of bacterial species and are involved in sensing environmental stimuli. We used two oligonucleotide probes matching the postulated coding regions for domains of sensor and regulator proteins respectively in Xanthomonas campestris pathovar campestris (Xcc) to identify possible two-component regulatory systems in Xcc. Two different fragments of Xcc DNA with homology to both of these probes were cloned. The DNA sequence of part of one of these fragments encompassed a potential open reading frame (ORF), the predicted amino acid sequence of which had extensive homology with regulator proteins of two-component regulatory systems. Analysis of the predicted amino acid sequence for the 3' end of an adjacent ORF revealed a very high level of homology with the C-terminal end of sensor proteins. Strains of Xcc with Tn5-induced mutations in the regulator gene were affected in extracellular polysaccharide production, and also in resistance to salt and chloramphenicol. No effects of mutation in the second clone were observed.

Amino Acid Sequence

Factors affecting the incidence and anti-salmonella activity of the anaerobic caecal flora of the young chick.

Thirty-two different types of anaerobic bacteria isolated from chickens have been tested for anti-salmonella activity in vitro. Under the conditions of the test only Bacteroides hypermegas and a Bifidobacterium sp. were shown to inhibit the salmonellas and this was attributed to the production of volatile fatty acids (VFA's) coupled with a low pH. When these organisms were tested in newly hatched chicks no inhibition of S. typhimurium occurred. Possible explanations for this observation are considered. The pH value and concentration of VFA's in the caecal material were determined in chicks from 0-84 days. In vitro tests with S. typhimurium indicated that, whilst the organism would be able to multiply at the pH and concentration of VFA's found during the first few days after hatching, the rapid increase in VFA concentration during the first 21 days would make this increasingly difficult. The significance of the developing caecal flora in relation to VFA production and pH is discussed. Because certain feed additives are known to influence the carriage of salmonellas, the sensitivity of various caecal anaerobes to these compounds was determined in vitro, generally at 1, 10 and 100 microgram/ml. The additives tested included flavomycin, furazolidone, nitrovin, tetracyline, tylosin, sulphaquinoxaline, virginiamycin and zinc bacitracin. All the organisms tested were inhibited by 100 microgram/ml furazolidone; none were inhibited by 500 microgram/ml sulphaquinoxaline. Changes occurring in the VFA concentration, pH value and microflora of the caeca of chicks fed for 49 days or longer on a normal starter diet or the same diet containing 10 or 100 mg/kg nitrovin have been compared. When the chicks were fed on the diet containing 100 mg/kg nitrovin, the Gram-negative non-sporing anaerobes were eliminated as a significant part of the caecal flora. However, the VFA concentration combined with a low pH in chicks from 2 weeks onwards was still sufficient to inhibit salmonella multiplication. Other possibly interrelated factors which might lead to an increased salmonella carrier rate in the nitrovin-treated chickens are discussed.

Anaerobiosis