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LF f(17-42) [id=LFB0092]

Synonym: Peptide I

Producer Organism : Native Protein : Production Method :
Cow Lactoferrin (LF) Enzymatic hydrolysis and Purification with LC method
Activity : Antibacterial
Target Organisms :

Gram-positive: Micrococcus flavus DSM 1790 (MIC=8 然), Staphylococcus epidermidis CECT 231 (MIC=0.05 然), Bacillus cereus ACM 446 (MIC=8 然), Listeria monocytogenes NCTC 7973 (MIC=2 然), Staphylococcus aureus NCTC 6571 (MIC=16 然).

Gram-negative: Escherichia coli ATCC 25922 (MIC=16 然), Escherichia coli ATCC 25922 (MIC=0.0125 然), Escherichia coli strain 1 (MIC=4 然), Pseudomonas fluorescens wild strain (MIC=16 然), Salmonella salford IMVS 1710 (MIC=8 然).

Description :
Production method: Porcin pepsin hydrolysis and purification with LC method.

LFB0012 shares structural homology with human lactoferricin (LFH0009) both peptides contain two subfragments linked by a disulfide bridge, and both have similar origins in the native molecule (1 to 47 and 1 to 48 for human and bovine peptides, respectively). LFB0012 was four times less active than LFB0092, even though LFB0012 contained the complete sequence of LFB0092. Others studies found that the activity of LFH0009 was nine times lower than that of LFB0084. It may be speculated that, in the larger peptides (LFH0009 and LFB0012), sequences flanking the active loop (residues 20 to 37 and 19 to 36 in human and bovine peptides, respectively) hinder effective interaction with the bacterial surface. With LFB0092, a molecule with smaller flanking sequences, such interference would be minimized (Citation: 5).
Length : 26 Mass (Da): 3 197.33 Common Amino Acids : R
Isolectric Point : 12.35 Net Charge : 8 Absent Amino Acids : DEHNY
Basic Residues : 8 Acidic Residues : 0 Hydrophobic Residues : 10
Polar Residues : 5 Boman Index : -66.98 Hydropathy Index : -0.485
Aliphatic Index : 52.69 Instability Index : 0 Extinction Coefficient : 11125
Absorbance 280nm : 445

Wheel representation

Hydrophobicity plot

Red solid plot : values according to the hydrophobicity scale of Kyte and Doolittle (reference paper).
Yellow dashed plot : Experimentally determined hydrophobicity scale for proteins at membrane interfaces(reference paper).
Green dotted-dashed plot : prediction of transmembrane helices (reference paper). In this scale (unlike the others), more negative values reflect greater hydrophobicity.

Multiple Sequence Alignment (MSA)


                      1 [        .         .         . ] 32
  1 LFBNATIVE 100.0%    FKCRRWQWRMKKLGAPSITCVRRAFALECIRA   
  2 LFB0017   100.0%    FKCRRWQWR-----------------------   
  3 LFB0043    93.3%    FKCRRWQWRAKKLGA-----------------   
  4 LFB0052    86.7%    FKCWRWQWRWKKLGA-----------------   
  5 LFB0037    93.3%    FKCARWQWRMKKLGA-----------------   
  6 LFB0040    93.3%    FKCRRWAWRMKKLGA-----------------   
  7 LFB0038    93.3%    FKCRAWQWRMKKLGA-----------------   
  8 LFB0088    96.0%    FKCRRWQWRMKKLGAPSITCVRRAE-------   
  9 LFB0039    93.3%    FKCRRAQWRMKKLGA-----------------   
 10 LFB0062    93.3%    FKCRRAQWRMKKLGA-----------------   
 11 LFB0072    93.3%    FKCRRAQWRMKKLGA-----------------   
 12 LFB0075    93.3%    FKCRRAQWRMKKLGA-----------------   
 13 LFB0078    93.3%    FKCRRAQWRMKKLGA-----------------   
 14 LFB0064    86.7%    FKCRRAQARMKKLGA-----------------   
 15 LFB0117    54.5%    ---RRAAARAKKAG------------------   
 16 LFB0080    86.7%    FKCRRAQARMKKLGA-----------------   
 17 LFB0074    86.7%    FKCRRAQARMKKLGA-----------------   
 18 LFB0077    86.7%    FKCRRAQARMKKLGA-----------------   
 19 LFB0056    93.3%    FKCRRFQWRMKKLGA-----------------   
 20 LFB0059    93.3%    FKCRRFQWRMKKLGA-----------------   
 21 LFB0053    93.3%    FKCRRFQWRMKKLGA-----------------   
 22 LFB0081    93.3%    FKCRRFQWRMKKLGA-----------------   
 23 LFB0055    86.7%    FKCRRFQFRMKKLGA-----------------   
 24 LFB0058    86.7%    FKCRRFQFRMKKLGA-----------------   
 25 LFB0061    86.7%    FKCRRFQFRMKKLGA-----------------   
 26 LFB0083    86.7%    FKCRRFQFRMKKLGA-----------------   
 27 LFB0041    93.3%    FKCRRWQARMKKLGA-----------------   
 28 LFB0063    93.3%    FKCRRWQARMKKLGA-----------------   
 29 LFB0076    93.3%    FKCRRWQARMKKLGA-----------------   
 30 LFB0079    93.3%    FKCRRWQARMKKLGA-----------------   
 31 LFB0073    93.3%    FKCRRWQARMKKLGA-----------------   
 32 LFB0060    93.3%    FKCRRWQFRMKKLGA-----------------   
 33 LFB0082    93.3%    FKCRRWQFRMKKLGA-----------------   
 34 LFB0057    93.3%    FKCRRWQFRMKKLGA-----------------   
 35 LFB0054    93.3%    FKCRRWQFRMKKLGA-----------------   
 36 LFB0042    93.3%    FKCRRWQWAMKKLGA-----------------   
 37 LFB0115    81.8%    ---RRWQRWMKKLG------------------   
 38 LFB0033   100.0%    FKCRRWQWRMKKLGA-----------------   
 39 LFB0032   100.0%    FKCRRWQWRMKKLGA-----------------   
 40 LFB0031   100.0%    FKCRRWQWRMKKLGA-----------------   
 41 LFB0050    86.7%    FKWRRWQWRMKKLWA-----------------   
 42 LFB0051    80.0%    FKWRRWWWRMKKLWA-----------------   
 43 LFB0049    93.3%    FKCRRWQWRMKKLWA-----------------   
 44 LFB0018   100.0%    FKCRRWQWRM----------------------   
 45 LFB0020   100.0%    FKCRRWQWRMK---------------------   
 46 LFB0045    93.3%    FKCRRWQWRMKALGA-----------------   
 47 LFB0044    93.3%    FKCRRWQWRMAKLGA-----------------   
 48 LFB0097   100.0%    ---RRWQWR-----------------------   
 49 LFB0030   100.0%    FKCRRWQWRMKKLG------------------   
 50 LFB0087   100.0%    FKCRRWQWRMKKLGAPSITCVRRAF-------   
 51 LFB0024    54.5%    YKAWRWAWRWK---------------------   
 52 LFB0025    54.5%    YKAWRWAWRWK---------------------   
 53 LFB0027    36.4%    YRMWRWAWRWR---------------------   
 54 LFB0028    36.4%    YRMWRWRWRWR---------------------   
 55 LFB0026    36.4%    YRAWRWAWRWR---------------------   
 56 LFB0023    63.6%    YKARRWAWRWK---------------------   
 57 LFB0022    72.7%    YKARRWAWRMK---------------------   
 58 LFB0021    81.8%    FKARRWAWRMK---------------------   
 59 LFB0019    90.0%    FKARRWQWRM----------------------   
 60 LFB0036    93.3%    FKARRWQWRMKKLGA-----------------   
 61 LFB0070    93.3%    FKARRWQWRMKKLGA-----------------   
 62 LFB0069    93.3%    FKARRWQWRMKKLGA-----------------   
 63 LFB0068    93.3%    FKARRWQWRMKKLGA-----------------   
 64 LFB0065    93.3%    FKARRWQWRMKKLGA-----------------   
 65 LFB0106   100.0%    ---RRWQWRMKK--------------------   
 66 LFB0029    45.5%    RRWYRWAWRMR---------------------   
 67 LFB0118   100.0%    ----RWQWRM----------------------   
 68 LFB0113    81.8%    ---RRWQWRMRRLG------------------   
 69 LFB0112    72.7%    ---KKWQWKMKKLG------------------   
 70 LFB0114    81.8%    ---KKWQWRMKKLG------------------   
 71 LFB0116    54.5%    ---EEWQWEMEELG------------------   
 72 LFB0048    93.3%    FKWRRWQWRMKKLGA-----------------   
 73 LFB0090    92.0%    FKSRRWQWRMKKLGAPSITSVRRAF-------   
 74 LFB0110    18.2%    ----RRWQWRMKKLG-----------------   
 75 LFB0067    93.3%    FKFRRWQWRMKKLGA-----------------   
 76 LFB0071    93.3%    FKFRRWQWRMKKLGA-----------------   
 77 LFB0066    93.3%    FKFRRWQWRMKKLGA-----------------   
 78 LFB0108   100.0%    ---RRWQWRMKKL-------------------   
 79 LFB0102   100.0%    ---RRWQWRMKK--------------------   
 80 LFB0034    93.3%    AKCRRWQWRMKKLGA-----------------   
 81 LFB0111   100.0%    ---RRWQWRMKKLG------------------   
 82 LFB0104   100.0%    ---RRWQWRMKK--------------------   
 83 LFB0035    93.3%    FACRRWQWRMKKLGA-----------------   
 84 LFB0109   100.0%    ---RRWQWRMKKLG------------------   
 85 LFB0101   100.0%    ---RRWQWRMKK--------------------   
 86 LFB0046    93.3%    FKCRRWQWRMKKAGA-----------------   
 87 LFB0107   100.0%    ---RRWQWRMKK--------------------   
 88 LFB0100   100.0%    ---RRWQWRMKK--------------------   
 89 LFB0047    93.3%    FKCRRWQWRMKKLAA-----------------   
 90 LFB0098   100.0%    ---RRWQWRMKK--------------------   
 91 LFB0105   100.0%    ---RRWQWRMKK--------------------   
 92 LFB0103   100.0%    ---RRWQWRMKK--------------------   
 93 LFB0086   100.0%    FKCRRWQWRMKKLGAPSITCVRRAF-------   
 94 LFB0089   100.0%    FKCRRWQWRMKKLGAPSITCVRRAF-------   
 95 LFB0085   100.0%    FKCRRWQWRMKKLGAPSITCVRRAF-------   
 96 LFB0084   100.0%    FKCRRWQWRMKKLGAPSITCVRRAF-------   
 97 LFB0092   100.0%    FKCRRWQWRMKKLGAPSITCVRRAFA------   
 98 LFB0095   100.0%    -KCRRWQWRMKKLGAPSITCV-----------   
 99 LFB0096   100.0%    --CRRWQWRMKKLGAPSITCV-----------   

100 LFB0093   100.0%    FKCRRWQWRMKKLGAPSITCVRRAFAL-----   

101 LFB0091   100.0%    FKCRRWQWRMKKLGAPSITCVRRAFA------   

102 LFB0119   100.0%    ----------KKLGAPSITCVRRAFA------   
103 LFB0120   100.0%    -------------GAPSITCVRRAF-------   

104 LFB0121   100.0%    ----------------------------CIRA   
105 LFB0094   100.0%    FKCRRWQWRMKKLGAPSITCVRRAFALECIR-   

Citation: 1

Synergistic effect between different milk-derived peptides and proteins

Cited Entries: LFB0085, LFB0092, CAA0021

Authors:Lopez-Exposito, I., Pellegrini, A., Amigo, L., Recio, I.
Journal: Journal of Dairy Science 2008, 91(6).
CrossRef External Link
Abstract: Antimicrobial peptides derived from food proteins constitute a new field in the combined use of antimicrobial agents in food. The best examples of milk-derived peptides are those constituted by bovine lactoferricin [lactoferrin f(1741)] (LFcin-B) and bovine αs2-casein f(183207). The aim of this work was to study if the antimicrobial activity of a natural compound employed in food preservation, nisin, could be enhanced by combination with the aforementioned milk-derived peptides. Furthermore, the possibility of a synergistic effect between these peptides and bovine lactoferrin (LF) against Escherichia coli and Staphylococcus epidermidis was also studied. Finally, the most active combinations were assayed against the foodborne pathogens Listeria monocytogenes and Salmonella choleraesuis. Results showed a synergistic effect when LFcin-B was combined with bovine LF against E. coli. In the same way, the combination of LFcin-B with bovine LF was synergistic against Staph. epidermidis. Bovine LF and nisin increased their antimicrobial activity when they were assayed together with bovine αs2-casein f(183-207). It is important to note the synergistic effect among LFcin-B and bovine LF, because both compounds might be simultaneously in the suckling gastrointestinal tract and could, therefore, have a protective effect on it. The other synergistic effect high-lighted is that between αs2-casein f(183207) and nisin against L. monocytogenes because of the ability of L. monocytogenes to develop resistance to nisin.
Keywords: synergism; milk-derived antibacterial peptide; antibacterial milk protein
Citation: 2

Two ion-exchange chromatographic methods for the isolation of antibacterial peptides from lactoferrin: In situ enzymatic hydrolysis on an ion-exchange membrane

Cited Entries: LFB0006, LFB0010, LFB0085, LFB0092, LFB0147, LFB0148, LFB0151, CAA0013, CAA0019, CAA0020

Authors:Recio, I., Visser, S.
Journal: Journal of Chromatography A 1999, 831(2).
CrossRef External Link
Abstract: Two ion-exchange chromatographic methods are reported for the rapid isolation of antibacterial peptides from lactoferrin (LF). Using the first method, a pepsin hydrolysate of LF was fractionated by bead-based cation-exchange chromatography. After removal of weakly bound material by washing with ammonia, highly purified lactoferricin-B (LFcin-B) was obtained in a single step by elution with 2 M NaCl. Some other cationic peptides, copurified as minor components, were also characterised by N-terminal sequencing, mass spectrometry and antibacterial activity determination. With the second method, cheese whey was filtered through a cation-exchange membrane, and the selectively bound LF was directly hydrolysed in situ with pepsin. Inactive LF fragments were washed off the membrane with ammonia, and a fraction enriched in LFcin-B was obtained by further elution with 2 M NaCl. The membrane method is more rapid and offers several economic advantages.
Keywords: peptides; ANTIBACTERIAL agents; Lactoferrin
Citation: 3

Antibacterial and binding characteristics of bovine, ovine and caprine lactoferrins: a comparative study

Cited Entries: LFB0085, LFB0092, LAG0001, LAG0014

Authors:Recio, I., Visser, S.
Journal: International Dairy Journal 2000, 10(9).
CrossRef External Link
Abstract: A cation-exchange chromatographic membrane was used for the rapid isolation of caprine and ovine lactoferrin (LF) from cheese whey. Caprine LF was isolated by one-step cation-exchange chromatography, but the isolation of ovine LF needed a further reversed-phase HPLC purification step. The antibacterial activity of these LFs, determined towards Micrococcus flavus and Escherichia coli, was compared with that of bovine LF. Of the apo-LFs the caprine protein had the highest activity. In all cases the holo-LFs showed low or negligible activity. The antibacterial properties of cationic peptides obtained by peptic hydrolysis of caprine LF were studied against E. coli and M. flavus, and the activity of these peptides was compared with that of bovine lactoferricin. In addition, the binding characteristics of bovine, ovine and caprine LF to bacterial cells of E. coli were investigated with an enzyme-linked binding assay using horseradish peroxidase-bovine LF as a conjugate. Ovine and bovine LF strongly inhibited the binding of the LF complex to the bacterial surface. The iron-free forms of these LFs showed a greater ability for binding to the bacterial cells than the iron-saturated forms. However, several cationic peptides and caprine LF did not inhibit the binding of the LF conjugate although they exhibited a marked antibacterial effect. The results of binding of the LF complex in the presence of LFs from different species and of membrane-active peptides are discussed in relation to the antibacterial activity of these proteins and peptides.
Keywords: Lactoferrin; bovine; Ovine; Caprine; Antibacterial peptides; Antibacterial activity; Bacterial cell binding
Citation: 4

Identification of the initial binding sites of alphas2-casein f(183-207) and effect on bacterial membranes and cell morphology

Cited Entries: LFB0085, LFB0092, CAA0021

Authors:Lopez-Exposito, I., Amigo, L., Recio, I.
Journal: Biochimica et Biophysica Acta (BBA) - Biomembranes 2008, 1778(10).
CrossRef External Link
Abstract: The aim of this work was to identify the initial binding sites to the bacterial membranes of the antimicrobial peptide αs2-casein f(183207) and also to acquire further insight into membrane permeabilization of this peptide. Furthermore, cell morphology was studied by transmission electron microscopy. In all the experiments, bovine LFcin was employed as a comparison. Results showed that initial binding sites of αs2-casein f(183207) peptide were lipoteichoic acid in Gram-positive bacteria and lipopolysaccharide in Gram-negative. The peptide was able to permeabilize the outer and inner membranes. Moreover, the αs2-casein peptide f(183207) generated pores in the outer membrane of Gram-negative bacteria and in the cell wall of Gram-positive bacteria. In the Gram-negative bacteria, f(183207) originated cytoplasm condensation, and in the Gram-positive bacteria the cytoplasmic content leaked into the extracellular medium. Furthermore, the experiments of inner and outer membrane permeabilization performed with LFcin-B showed that this peptide also has the ability to permeabilize both the inner and outer membranes.
Keywords: Antimicrobial peptides; Binding sites; Permeabilization; [alpha]s2-Casein; Milk-derived bioactive peptides
Citation: 5

Antibacterial peptides of bovine lactoferrin: purification and characterization

Cited Entries: LFB0009, LFB0012, LFB0092

Authors:Dionysius, D.A., Milne, J.M.
Journal: Journal of Dairy Science 1997, 80(4).
CrossRef External Link
Abstract: Three peptides with antibacterial activity toward enterotoxigenic Escherichia coli have been purified from a pepsin digest of bovine lactoferrin. All peptides were cationic and originated from the N-terminus of the molecule in a region where a bactericidal peptide, lactoferricin B, had been previously identified. The most potent peptide, peptide I, was almost identical to lactoferricin B; the sequence corresponded to residues 17 to 42, and the molecular mass was 3195 as deter-mined by mass spectrometry. A second, less active peptide, peptide II, consisted of two sequences, residues 1 to 16 and 43 to 48 (molecular mass of 2673), linked by a single disulfide bond. The third peptide, peptide III, also a disulfide-linked heter-odimer, corresponded to residues 1 to 48 (molecular mass of 5851), cleaved between residues 42 and 43. Peptides I and II displayed antibacterial activity toward a number of pathogenic and food spoilage microorganisms, and peptide I inhibited the growth of Listeria monocytogenes at concentrations as low as 2 然. Bacterial growth curves showed that bactericidal effects of peptides I and II were observable within 30 min of exposure. The results confirmed and extended those of earlier studies suggesting that the bactericidal domain of lactoferrin was localized in the N-terminus and did not involve iron-binding sites.
Keywords: Lactoferrin; pepsin digest; Antibacterial peptides

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