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Title: Photothermal spectroscopy of Bacillus anthracis and Bacillus cereus with microcantilevers

Abstract

Microcalorimetric optical and infrared spectroscopy is a method of determining the spectral absorption of small quantities of materials over a wide range of incident wavelengths. In this paper, the first spectroscopic results for microcantilevers coated with Bacillus anthracis (BA) are presented. These results, for B. anthracis from 2.5 to 14.5 {micro}m, are compared with results from microcantilevers coated with Bacillus cereus (BC) and standard spectroscopic absorption data. The results demonstrate strong correlation between the deflection measurements and the reference spectroscopic absorption peaks. An advantage of this microcantilever-based method over traditional spectroscopy is that much smaller amounts of material (nanogram quantities) can be detected in comparison with the milligram amounts needed for standard methods. Another advantage is that the complete system can be relatively small without sacrificing spectral resolution.

Authors:
 [1];  [1];  [1];  [1];  [1]
  1. ORNL
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
989539
DOE Contract Number:
DE-AC05-00OR22725
Resource Type:
Journal Article
Resource Relation:
Journal Name: Sensors and Actuators B: Chemical; Journal Volume: 114; Journal Issue: 1
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; ABSORPTION; BACILLUS; BACILLUS CEREUS; RESOLUTION; SPECTROSCOPY; WAVELENGTHS; microcantilever; sensor; photothermal

Citation Formats

Wig, Andrew G, Arakawa, Edward T, Passian, Ali, Ferrell, Thomas L, and Thundat, Thomas George. Photothermal spectroscopy of Bacillus anthracis and Bacillus cereus with microcantilevers. United States: N. p., 2006. Web. doi:10.1016/j.snb.2005.04.029.
Wig, Andrew G, Arakawa, Edward T, Passian, Ali, Ferrell, Thomas L, & Thundat, Thomas George. Photothermal spectroscopy of Bacillus anthracis and Bacillus cereus with microcantilevers. United States. doi:10.1016/j.snb.2005.04.029.
Wig, Andrew G, Arakawa, Edward T, Passian, Ali, Ferrell, Thomas L, and Thundat, Thomas George. Wed . "Photothermal spectroscopy of Bacillus anthracis and Bacillus cereus with microcantilevers". United States. doi:10.1016/j.snb.2005.04.029.
@article{osti_989539,
title = {Photothermal spectroscopy of Bacillus anthracis and Bacillus cereus with microcantilevers},
author = {Wig, Andrew G and Arakawa, Edward T and Passian, Ali and Ferrell, Thomas L and Thundat, Thomas George},
abstractNote = {Microcalorimetric optical and infrared spectroscopy is a method of determining the spectral absorption of small quantities of materials over a wide range of incident wavelengths. In this paper, the first spectroscopic results for microcantilevers coated with Bacillus anthracis (BA) are presented. These results, for B. anthracis from 2.5 to 14.5 {micro}m, are compared with results from microcantilevers coated with Bacillus cereus (BC) and standard spectroscopic absorption data. The results demonstrate strong correlation between the deflection measurements and the reference spectroscopic absorption peaks. An advantage of this microcantilever-based method over traditional spectroscopy is that much smaller amounts of material (nanogram quantities) can be detected in comparison with the milligram amounts needed for standard methods. Another advantage is that the complete system can be relatively small without sacrificing spectral resolution.},
doi = {10.1016/j.snb.2005.04.029},
journal = {Sensors and Actuators B: Chemical},
number = 1,
volume = 114,
place = {United States},
year = {Wed Mar 01 00:00:00 EST 2006},
month = {Wed Mar 01 00:00:00 EST 2006}
}
  • No abstract prepared.
  • No abstract prepared.
  • The genomes of Bacillus cereus and its closest relative Bacillus anthracis each contain two LmbE protein family homologs: BC1534 (BA1557) and BC3461 (BA3524). Only a few members of this family have been biochemically characterized including N-acetylglucosaminylphosphatidyl inositol (GlcNAc-PI), 1- D-myo-inosityl-2-acetamido-2-deoxy-α- D-glucopyranoside (GlcNAc-Ins), N,N'-diacetylchitobiose (GlcNAc 2) and lipoglycopeptide antibiotic de-N-acetylases. All these enzymes share a common feature in that they de-N-acetylate the N-acetyl- D-glucosamine (GlcNAc) moiety of their substrates. The bc1534 gene has previously been cloned and expressed in Escherichia coli. The recombinant enzyme was purified and its 3D structure determined. In this study, the bc3461 gene from B. cereus ATCC14579more » was cloned and expressed in E. coli. The recombinant enzymes BC1534 (EC 3.5.1.-) and BC3461 were biochemically characterized. The enzymes have different molecular masses, pH and temperature optima and broad substrate specificity, de-N-acetylating GlcNAc and N-acetylchito-oligomers (GlcNAc 2, GlcNAc 3 and GlcNAc 4), as well as GlcNAc-1P, N-acetyl-d-glucosamine-1 phosphate; GlcNAc-6P, N-acetyl-d-glucosamine-6 phosphate; GalNAc, N-acetyl-d-galactosamine; ManNAc, N-acetyl-d-mannosamine; UDP-GlcNAc, uridine 5'-diphosphate N-acetyl-d-glucosamine. However, the enzymes were not active on radiolabeled glycol chitin, peptidoglycan from B. cereus, N-acetyl-d-glucosaminyl-(β-1,4)-N-acetylmuramyl-l-alanyl-d-isoglutamine (GMDP) or N-acetyl-d-GlcN-Nα1-6-d-myo-inositol-1-HPO 4-octadecyl (GlcNAc-I-P-C 18). Kinetic analysis of the activity of BC1534 and BC3461 on GlcNAc and GlcNAc 2 revealed that GlcNAc 2 is the favored substrate for both native enzymes. Based on the recently determined crystal structure of BC1534, a mutational analysis identified functional key residues, highlighting their importance for the catalytic mechanism and the substrate specificity of the enzyme. Finally, the catalytic efficiencies of BC1534 variants were significantly decreased compared to the native enzyme. An alignment-based tree places both de-N-acetylases in functional categories that are different from those of other LmbE proteins.« less
  • The sequencing and analysis of two close relatives of Bacillus anthracis are reported. AFLP analysis of over 300 isolates of B.cereus, B. thuringiensis and B. anthracis identified two isolates as being very closely related to B. anthracis. One, a B. cereus, BcE33L, was isolated from a zebra carcass in Nambia; the second, a B. thuringiensis, 97-27, was isolated from a necrotic human wound. The B. cereus appears to be the closest anthracis relative sequenced to date. A core genome of over 3,900 genes was compiled for the Bacillus cereus group, including Banthracis. Comparative analysis of these two genomes with othermore » members of the B. cereus group provides insight into the evolutionary relationships among these organisms. Evidence is presented that differential regulation modulates virulence, rather than simple acquisition of virulence factors. These genome sequences provide insight into the molecular mechanisms contributing to the host range and virulence of this group of organisms.« less