藉基因工程產製天然抗菌物質乳鐵蛋白素(2/2)
Date Issued
2004
Date
2004
Author(s)
林慶文
DOI
922313B002134
Abstract
Bovine lactoferrin is a transferrin protein found
in milk, which has many biological functions.
Lactoferricin is a short fragment from the
N-terminus of lactoferrin. This peptide is about
25 amino acids. Lactoferricin is generated upon
gastric pepsin cleavage of lactoferrin in
mammalian animals. This peptide is much
more effective in antibacterial properties than
lactoferrin. And we could use the peptide to
develop new style probiotic products. Our
study is focus on the generation of the
recombinant yeast, Pichia pastoris, containing
the bovine lactoferricin gene fragment and the
feasibility of large-scale production. First, we
got synthetic lactoferricin and it’s antibacterial
core peptide of different species by peptide
synthesis, and then tested the antimicrobial
activities against Escherichia coli, Staphylococcus
aureus and Candida albicans. Results
revealed that the bovine lactoferricin and its
antibacterial core peptide show the better
ability of antimicrobial than the others. Then
11-amino acids antibacterial core peptide of
bovine lactoferricin were selected, which has
the same antibacterial activity as the native
lactoferricin. Using oligonucleotide synthesis
to get oligonucleotides of desired peptides, and
cloned the target gene into yeast-expression
vectors. After the constructed bLFcin/pPICZα
B vectors were confirmed, we transformed the
recombinant vectors into Pichia pastoris by
electroporation and selected the transformants
to express the biological antibacterial peptides.
In protein analysis, an expected protein size of
recombinant bLFcin was detected by Tricine
SDS-PAGE and Western blot. The results
revealed that recombinant bLFcin was
successful produced by P. pastoris expression
system.
Subjects
Lactoferricin
antibacterial peptide
Pichia pastoris
yeast expression cassette
Publisher
臺北市：國立臺灣大學動物科學技術學系暨研究所
Type
report
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