Low molecular weight chitosans—Preparation with the aid of pronase, characterization and their bactericidal activity towards Bacillus cereus and Escherichia coli
The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5–8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5, 37 °C) of chitosan showed lyses of Bacillus cereus and Escherichia coli more efficiently (100%) than native chitosan (< 50%). IR and 1H-NM...
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| Published in | Biochimica et biophysica acta Vol. 1770; no. 4; pp. 495 - 505 |
|---|---|
| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
Netherlands
Elsevier B.V
01.04.2007
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| Abstract | The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5–8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5, 37 °C) of chitosan showed lyses of
Bacillus cereus and
Escherichia coli more efficiently (100%) than native chitosan (<
50%). IR and
1H-NMR data showed decrease in the degree of acetylation (14–19%) in LMWC compared to native chitosan (∼
26%). Minimum inhibitory concentration of LMWC towards 10
6 CFU ml
−
1
of
B. cereus was 0.01% (w/v) compared to 0.03% for 10
4 CFU ml
−
1
of
E. coli. SEM revealed pore formation as well as permeabilization of the bacterial cells, as also evidenced by increased carbohydrate and protein contents as well as the cytoplasmic enzymes in the cell-free supernatants. N-terminal sequence analyses of the released proteins revealed them to be cytoplasmic/membrane proteins. Upon GLC, the supernatant showed characteristic fatty acid profiles in
E. coli, thus subscribing to detachment of lipopolysaccharides into the medium, whereas that of
B. cereus indicated release of surface lipids. The mechanism for the observed bactericidal activity of LMWC towards both Gram-positive and Gram-negative bacteria has been discussed. |
|---|---|
| AbstractList | The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5-8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5, 37 degrees C) of chitosan showed lyses of Bacillus cereus and Escherichia coli more efficiently (100%) than native chitosan (<50%). IR and (1)H-NMR data showed decrease in the degree of acetylation (14-19%) in LMWC compared to native chitosan ( approximately 26%). Minimum inhibitory concentration of LMWC towards 10(6) CFU ml(-1) of B. cereus was 0.01% (w/v) compared to 0.03% for 10(4) CFU ml(-1) of E. coli. SEM revealed pore formation as well as permeabilization of the bacterial cells, as also evidenced by increased carbohydrate and protein contents as well as the cytoplasmic enzymes in the cell-free supernatants. N-terminal sequence analyses of the released proteins revealed them to be cytoplasmic/membrane proteins. Upon GLC, the supernatant showed characteristic fatty acid profiles in E. coli, thus subscribing to detachment of lipopolysaccharides into the medium, whereas that of B. cereus indicated release of surface lipids. The mechanism for the observed bactericidal activity of LMWC towards both Gram-positive and Gram-negative bacteria has been discussed.The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5-8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5, 37 degrees C) of chitosan showed lyses of Bacillus cereus and Escherichia coli more efficiently (100%) than native chitosan (<50%). IR and (1)H-NMR data showed decrease in the degree of acetylation (14-19%) in LMWC compared to native chitosan ( approximately 26%). Minimum inhibitory concentration of LMWC towards 10(6) CFU ml(-1) of B. cereus was 0.01% (w/v) compared to 0.03% for 10(4) CFU ml(-1) of E. coli. SEM revealed pore formation as well as permeabilization of the bacterial cells, as also evidenced by increased carbohydrate and protein contents as well as the cytoplasmic enzymes in the cell-free supernatants. N-terminal sequence analyses of the released proteins revealed them to be cytoplasmic/membrane proteins. Upon GLC, the supernatant showed characteristic fatty acid profiles in E. coli, thus subscribing to detachment of lipopolysaccharides into the medium, whereas that of B. cereus indicated release of surface lipids. The mechanism for the observed bactericidal activity of LMWC towards both Gram-positive and Gram-negative bacteria has been discussed. The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5-8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5, 37 degrees C) of chitosan showed lyses of Bacillus cereus and Escherichia coli more efficiently (100%) than native chitosan (<50%). IR and (1)H-NMR data showed decrease in the degree of acetylation (14-19%) in LMWC compared to native chitosan ( approximately 26%). Minimum inhibitory concentration of LMWC towards 10(6) CFU ml(-1) of B. cereus was 0.01% (w/v) compared to 0.03% for 10(4) CFU ml(-1) of E. coli. SEM revealed pore formation as well as permeabilization of the bacterial cells, as also evidenced by increased carbohydrate and protein contents as well as the cytoplasmic enzymes in the cell-free supernatants. N-terminal sequence analyses of the released proteins revealed them to be cytoplasmic/membrane proteins. Upon GLC, the supernatant showed characteristic fatty acid profiles in E. coli, thus subscribing to detachment of lipopolysaccharides into the medium, whereas that of B. cereus indicated release of surface lipids. The mechanism for the observed bactericidal activity of LMWC towards both Gram-positive and Gram-negative bacteria has been discussed. The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5–8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5, 37 °C) of chitosan showed lyses of Bacillus cereus and Escherichia coli more efficiently (100%) than native chitosan (< 50%). IR and 1H-NMR data showed decrease in the degree of acetylation (14–19%) in LMWC compared to native chitosan (∼ 26%). Minimum inhibitory concentration of LMWC towards 10 6 CFU ml − 1 of B. cereus was 0.01% (w/v) compared to 0.03% for 10 4 CFU ml − 1 of E. coli. SEM revealed pore formation as well as permeabilization of the bacterial cells, as also evidenced by increased carbohydrate and protein contents as well as the cytoplasmic enzymes in the cell-free supernatants. N-terminal sequence analyses of the released proteins revealed them to be cytoplasmic/membrane proteins. Upon GLC, the supernatant showed characteristic fatty acid profiles in E. coli, thus subscribing to detachment of lipopolysaccharides into the medium, whereas that of B. cereus indicated release of surface lipids. The mechanism for the observed bactericidal activity of LMWC towards both Gram-positive and Gram-negative bacteria has been discussed. |
| Author | Varadaraj, Mandyam C. Vishu Kumar, Acharya B. Gowda, Lalitha R. Tharanathan, Rudrapatnam N. |
| Author_xml | – sequence: 1 givenname: Acharya B. surname: Vishu Kumar fullname: Vishu Kumar, Acharya B. organization: Department of Biochemistry and Nutrition, Central Food Technological Research Institute, Mysore-570020, India – sequence: 2 givenname: Mandyam C. surname: Varadaraj fullname: Varadaraj, Mandyam C. organization: Human Resource Development, Central Food Technological Research Institute, Mysore-570020, India – sequence: 3 givenname: Lalitha R. surname: Gowda fullname: Gowda, Lalitha R. organization: Protein Chemistry and Technology, Central Food Technological Research Institute, Mysore-570020, India – sequence: 4 givenname: Rudrapatnam N. surname: Tharanathan fullname: Tharanathan, Rudrapatnam N. email: tharanathan@yahoo.co.uk organization: Department of Biochemistry and Nutrition, Central Food Technological Research Institute, Mysore-570020, India |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/17240531$$D View this record in MEDLINE/PubMed |
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| Keywords | MIC DP LPS Bactericidal activity Mechanism CFU HPSEC Low molecular weight chitosan Pronase SEM GPC Structure DA M r GLC |
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| Snippet | The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5–8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5,... The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5-8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5,... |
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| SubjectTerms | Acetylation Amino Acid Sequence Anti-Bacterial Agents - chemistry Anti-Bacterial Agents - metabolism Anti-Bacterial Agents - pharmacology Bacillus cereus - drug effects Bacillus cereus - growth & development Bacillus cereus - ultrastructure Bactericidal activity Cell Membrane - drug effects Cell Membrane Permeability - drug effects Chitosan - chemistry Chitosan - metabolism Chitosan - pharmacology Chromatography, Gas Escherichia coli - drug effects Escherichia coli - growth & development Escherichia coli - ultrastructure Low molecular weight chitosan Magnetic Resonance Spectroscopy Mechanism Microbial Sensitivity Tests Microscopy, Electron, Scanning Models, Molecular Molecular Sequence Data Molecular Weight Pronase Pronase - chemistry Pronase - metabolism Protein Conformation Solubility Spectrophotometry, Infrared Structure |
| Title | Low molecular weight chitosans—Preparation with the aid of pronase, characterization and their bactericidal activity towards Bacillus cereus and Escherichia coli |
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