title: Preparation and antimicrobial action of three tryptic digested functional molecules of bovine lactoferrin.
abstract: Lactoferrin is an 80 kDa bilobal, iron binding glycoprotein which is primarily antimicrobial in nature. The hydrolysis of lactoferrin by various proteases in the gut produces several functional fragments of lactoferrin which have varying molecular sizes and properties. Here, bovine lactoferrin has been hydrolyzed by trypsin, the major enzyme present in the gut, to produce three functional molecules of sizes approximately 21 kDa, 38 kDa and 45 kDa. The molecules have been purified using ion exchange and gel filtration chromatography and identified using N-terminal sequencing, which reveals that while the 21 kDa molecule corresponds to the N2 domain (21LF), the 38 kDa represents the whole C-lobe (38LF) and the 45 kDa is a portion of N1 domain of N-lobe attached to the C-lobe (45LF). The iron binding and release properties of 21LF, 38LF and 45LF have been studied and compared. The sequence and structure analysis of the portions of the excision sites of LF from various species have been done. The antibacterial properties of these three molecules against bacterial strains, Streptococcus pyogenes, Escherichia coli, Yersinia enterocolitica and Listeria monocytogenes were investigated. The antifungal action of the molecules was also evaluated against Candida albicans. This is the first report on the antimicrobial actions of the trypsin cleaved functional molecules of lactoferrin from any species.
authors.affiliation: Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Nilisha
authors.initials: N
authors.lastname: Rastogi
authors.affiliation: Department of Microbiology, University of Delhi, South Campus, New Delhi, India.
authors.forename: Nitish
authors.initials: N
authors.lastname: Nagpal
authors.affiliation: Department of Biosciences, Jamia Milia Islamia, New Delhi, India.
authors.forename: Hammad
authors.initials: H
authors.lastname: Alam
authors.affiliation: Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Sadanand
authors.initials: S
authors.lastname: Pandey
authors.affiliation: Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Lovely
authors.initials: L
authors.lastname: Gautam
authors.affiliation: Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Mau
authors.initials: M
authors.lastname: Sinha
authors.affiliation: Food Science & Technology Institute, Morinaga Milk Industry Co. Ltd., Zama, Kanagawa, Japan.
authors.forename: Kouichirou
authors.initials: K
authors.lastname: Shin
authors.affiliation: Department of Biosciences, Jamia Milia Islamia, New Delhi, India.
authors.forename: Nikhat
authors.initials: N
authors.lastname: Manzoor
authors.affiliation: Department of Microbiology, University of Delhi, South Campus, New Delhi, India.
authors.forename: Jugsharan S
authors.initials: JS
authors.lastname: Virdi
authors.affiliation: Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Punit
authors.initials: P
authors.lastname: Kaur
authors.affiliation: Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Sujata
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Tej P
authors.initials: TP
authors.lastname: Singh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000378:metabolism / Q000494:pharmacology*; D002417:Cattle; D002852:Chromatography, Ion Exchange; D006863:Hydrogen-Ion Concentration; D007781:Lactoferrin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D010449:Peptide Mapping; D017386:Sequence Homology, Amino Acid; D014357:Trypsin / Q000378:metabolism*
chemical_list: D000890:Anti-Infective Agents; D010446:Peptide Fragments; D007781:Lactoferrin; D014357:Trypsin