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Construction, characterization and exemplificative application of bioluminescent Bifidobacterium longum biovar longum.
Int J Food Microbiol. 2008 Jun 10; 124(3):285-90.IJ

Abstract

The aim of this work was to construct a bifidobacterial biosensor that could be used to analyze the metabolic state of cells. We transformed by electroporation the human intestinal bacterium Bifidobacterium longum biovar longum with a vector (pGBL8b) containing the insect luciferase gene from a click beetle (Pyrophorus plagiophthalamus) and studied the basic parameters affecting light production in the bioluminescent phenotype. We detected a minimum of 4000 cells, which indicates that the insect luciferase expression in Bifidobacterium longum is extremely good, and a measurement requires only a few minutes of incubation in ambient oxygen conditions. A pH of 7.0 was optimal for incorporating the substrate d-luciferin, and the substrate saturation effect occurred at 125 microM. We employed bioluminescent B. longum for a quick test of the efficacy of different carbohydrates to preserve cell physiology under acidic conditions. The prebiotic compounds Actilight and lactulose were the most active in preventing loss of intracellular ATP during incubation at pH 3. Glucose and inulin were less active, though still effective. In sum, our results show that bioluminescent B. longum, transformed with the pGBL8b plasmid, is a valuable tool for rapidly studying the physiological state of anaerobic bacterial cells under different environmental conditions.

Authors+Show Affiliations

Department of Food Science and Microbiology, University of Milan, Italy. simone.guglielmetti@unimi.itNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18471916

Citation

Guglielmetti, Simone, et al. "Construction, Characterization and Exemplificative Application of Bioluminescent Bifidobacterium Longum Biovar Longum." International Journal of Food Microbiology, vol. 124, no. 3, 2008, pp. 285-90.
Guglielmetti S, Ciranna A, Mora D, et al. Construction, characterization and exemplificative application of bioluminescent Bifidobacterium longum biovar longum. Int J Food Microbiol. 2008;124(3):285-90.
Guglielmetti, S., Ciranna, A., Mora, D., Parini, C., & Karp, M. (2008). Construction, characterization and exemplificative application of bioluminescent Bifidobacterium longum biovar longum. International Journal of Food Microbiology, 124(3), 285-90. https://doi.org/10.1016/j.ijfoodmicro.2008.03.033
Guglielmetti S, et al. Construction, Characterization and Exemplificative Application of Bioluminescent Bifidobacterium Longum Biovar Longum. Int J Food Microbiol. 2008 Jun 10;124(3):285-90. PubMed PMID: 18471916.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Construction, characterization and exemplificative application of bioluminescent Bifidobacterium longum biovar longum. AU - Guglielmetti,Simone, AU - Ciranna,Alessandro, AU - Mora,Diego, AU - Parini,Carlo, AU - Karp,Matti, Y1 - 2008/04/03/ PY - 2008/02/27/received PY - 2008/03/25/revised PY - 2008/03/25/accepted PY - 2008/5/13/pubmed PY - 2008/8/19/medline PY - 2008/5/13/entrez SP - 285 EP - 90 JF - International journal of food microbiology JO - Int J Food Microbiol VL - 124 IS - 3 N2 - The aim of this work was to construct a bifidobacterial biosensor that could be used to analyze the metabolic state of cells. We transformed by electroporation the human intestinal bacterium Bifidobacterium longum biovar longum with a vector (pGBL8b) containing the insect luciferase gene from a click beetle (Pyrophorus plagiophthalamus) and studied the basic parameters affecting light production in the bioluminescent phenotype. We detected a minimum of 4000 cells, which indicates that the insect luciferase expression in Bifidobacterium longum is extremely good, and a measurement requires only a few minutes of incubation in ambient oxygen conditions. A pH of 7.0 was optimal for incorporating the substrate d-luciferin, and the substrate saturation effect occurred at 125 microM. We employed bioluminescent B. longum for a quick test of the efficacy of different carbohydrates to preserve cell physiology under acidic conditions. The prebiotic compounds Actilight and lactulose were the most active in preventing loss of intracellular ATP during incubation at pH 3. Glucose and inulin were less active, though still effective. In sum, our results show that bioluminescent B. longum, transformed with the pGBL8b plasmid, is a valuable tool for rapidly studying the physiological state of anaerobic bacterial cells under different environmental conditions. SN - 0168-1605 UR - https://www.unboundmedicine.com/medline/citation/18471916/Construction_characterization_and_exemplificative_application_of_bioluminescent_Bifidobacterium_longum_biovar_longum_ DB - PRIME DP - Unbound Medicine ER -