ABSTRACT
Recently, we reported the synthesis of five bacteriocin-like inhibitor substances (Bt-BLIS: morricin 269, kurstacin 287, kenyacin 404, entomocin 420, and tolworthcin 524) by Mexican strains of Bacillus thuringiensis. Here we show that, collectively, these Bt-BLIS have a moderate to broad spectrum of antibacterial activity, being toxic to clinically significant against Gram-positive and Gram-negative bacteria, including common etiological agents of human diseases, such as strep throat and scarlet fever, septicemia, pneumonia, urinary tract infection, and emetic and gastrointestinal syndromes. Although synthesis of the five Bt-BLIS was independent of the presence of a target inducing bacterium, we demonstrated for the first time that a proteinaceous component(s) secreted by, or liberated by proteolytic cleavage of Bacillus cereus 183 following treatment with proteinase K, enhanced Bt-BLIS synthesis.
Subject(s)
Anti-Bacterial Agents/biosynthesis , Anti-Bacterial Agents/pharmacology , Bacillus thuringiensis/metabolism , Bacteriocins/biosynthesis , Bacteriocins/pharmacology , Bacillus cereus/drug effects , Bacillus thuringiensis/classification , Food Microbiology , Fungi/drug effects , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Mexico , Microbial Sensitivity TestsABSTRACT
AIMS: To synthesize two heterologous endochitinases in Escherichia coli and demonstrate their potential for applied use in generating antibacterial chitin-derived oligosaccharides (OGS). METHODS AND RESULTS: Heterologous endochitinase genes, chiA Nima and chiA74, were expressed in E. coli. Endochitinases were secreted by the E. coli export machinery and by approximately 20 h maximal chitinolytic activity was observed. The highest chitinolytic activity was observed with ChiA Nima, which produced antibacterial OGS with activities against Enterobacter cloacae, Escherichia coli, Staphylococcus aureus and S. xylosus. CONCLUSIONS: It was shown that the export machinery of E. coli is well suited for the secretion of bioactive ChiA74 and ChiA Nima endochitinases, and that the latter can generate antibacterial OGS. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study suggests that it is feasible to synthesize endochitinases ChiA Nima and ChiA74 codified by E. coli and mass-produce these enzymes in culture supernatants. As signal peptides in native ChiA Nima and ChiA74 were recognized by the protein export molecular apparatus in E. coli, these short peptides could be included as signal sequences for transport in E. coli of other proteins with applied value. This is the first report suggesting that ChiA Nima can be used to produce OGS to control food-borne pathogenic bacteria.
Subject(s)
Anti-Bacterial Agents/metabolism , Chitinases/genetics , Chitinases/metabolism , Escherichia coli/enzymology , Oligosaccharides/metabolism , Amino Acid Sequence , Chitin/metabolism , DNA Primers , Electrophoresis, Polyacrylamide Gel/methods , Escherichia coli/genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The protein quality of fruits bodies of three Pleurotus ostreatus Mexican strains (INIREB-8, CDBB-H-896 and CDBB-H-897) was evaluated. The protein concentration (Nx4.38) ranged from 17.26 to 19.97 g/100 g dry weight; chemical scores were between 74 and 93% with available lysine as a first limiting amino acid in either INIREB-8 and CDBB-H-896 strains or leucine in CDBB-H-897 strain. The nutritional evaluation revealed 67.75 to 68.38% in vitro digestibility. Relative protein values were from 100.06-107.85%, which were lower than soybean meal and whole egg but larger than those of rice, maize, beans, fava beans and pasta, no differences were found between these values and those of skim milk powder, casein plus methionine and albumin. In accordance with the last results we concluded that due to their essential amino acids content, mushroom proteins are a good complement of cereals; furthermore, it is highly recommended to include Pleurotus in the daily diet.
Subject(s)
Plant Proteins/analysis , Plants, Edible/chemistry , Pleurotus/chemistry , Mexico , Nutritive ValueABSTRACT
The chemical composition of three Mexican strains of Pleurotus ostreatus (INIREB-8, CDBB-H-896 and CDBB-H-897), were determined. The mushrooms were cultivated on wheat straw in a greenhouse (22-28 degrees C temperature and 80 +/- 5% of relative humidity). Fruits bodies of P. ostreatus contained (all values are expressed in g/100 g dry wt.), protein (N x 6.25): 24.64 +/- 0.21-28.50 +/- 0.26; lipids: 1.10 +/- 0.16-1.85 +/- 0.22; mineral matter: 7.66 +/- 0.23-8.79 +/- 0.25; dietary fibre: 32.14 +/- 0.14-36.81 +/- 0.40; and available carbohydrates: 26.33 +/- 1.04-30.46 +/- 0.21. They contain vitamins (mg/100 g dry wt): riboflavin: 3.31-3.7, thiamin: 1.92-1.96, niacin: 35.98-36.56 and ascorbic acid: 28-35. The main fatty acid was linoleic (0.70-1.19 g/100 g dry wt), it was also reported a low calcium and phosphorus content. Concluding the Pleurotus ostreatus could be a source of some of the complex B vitamins, dietary fiber, protein and linoleic acid.
