Related Experiment Video
Updated: Feb 22, 2026
Antimicrobial Peptides Produced by Selective Pressure Incorporation of Non-canonical Amino Acids
Published on: May 4, 2018
Molecular cloning, recombinant expression and functional characterization of an antimicrobial peptide, Crustin from
K S Sruthy1, Aishwarya Nair1, J Puthumana2
1Department of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science and Technology, Fine Arts Avenue, Kochi 682016, Kerala, India.
Abstract:
Antimicrobial peptides (AMPs) comprise molecules that involve in the defense mechanism of various organisms towards pathogens such as bacteria, fungi, parasites and viruses. Crustins are generally defined as multi-domain cationic antimicrobial peptides containing one whey acidic protein (WAP) domain at the C-terminus as the functional unit. In this study, we identified and characterized a novel crustin homolog (Fi-Crustin2) with 354 bp fragment cDNA encoding 117 amino acids and an ORF of 100 amino acids with a net charge of +1 from the mRNA of F. indicus haemocytes. This study forms the second report of a crustin isoform from F. indicus. Blast analysis revealed that Fi-crustin2 exhibits similarity to shrimp crustins already reported. The active mature peptide has a molecular weight of 10.61 kDa and pI of 7.59 with a beta sheeted structure. The mature peptide was cloned into pET-32a(+) with a N-terminal hexa-histidine tag fused in-frame, and expressed in Escherichia coli, and the recombinant crustin, Fi-crustin2 inhibited the growth of Gram-negative bacteria with low MIC. All these features suggest that Fi-crustin2 is a potent antibacterial protein against Gram-negative bacteria and could play an important role in the innate immune mechanism of F. indicus.
Insights
Researchers identified a novel antimicrobial peptide, Fi-Crustin2, from the shrimp F. indicus. This potent antibacterial protein effectively inhibits Gram-negative bacteria, highlighting its role in the shrimp
Area of Science:
- Marine Biology
- Immunology
- Biochemistry
Background:
- Antimicrobial peptides (AMPs) are crucial for innate immunity in diverse organisms.
- Crustins are a class of AMPs characterized by a whey acidic protein (WAP) domain, important for antimicrobial activity.
- The immune mechanisms of the shrimp F. indicus are not fully understood.
Purpose of the Study:
- To identify and characterize a novel crustin homolog, Fi-Crustin2, from F. indicus haemocytes.
- To investigate the antibacterial activity of Fi-Crustin2 against various pathogens.
- To elucidate the potential role of Fi-Crustin2 in the innate immune system of F. indicus.
Main Methods:
- cDNA sequencing and analysis to identify Fi-Crustin2.
- Bioinformatic analysis including BLAST for sequence similarity.
- Recombinant expression of Fi-Crustin2 in Escherichia coli.
- Determination of antibacterial activity through Minimum Inhibitory Concentration (MIC) assays.
Main Results:
- A novel crustin homolog, Fi-Crustin2, was identified with a 354 bp cDNA fragment encoding 117 amino acids.
- Fi-Crustin2 shares similarity with known shrimp crustins.
- The recombinant Fi-Crustin2 exhibited potent inhibitory activity against Gram-negative bacteria with low MIC values.
- The mature peptide has a molecular weight of 10.61 kDa, a pI of 7.59, and a beta-sheet structure.
Conclusions:
- Fi-Crustin2 is a potent antibacterial protein effective against Gram-negative bacteria.
- This novel crustin plays a significant role in the innate immune defense of F. indicus.
- Further studies on Fi-Crustin2 could lead to new antimicrobial strategies.
More Related Videos
12:23Recombinant Protein Expression, Crystallization, and Biophysical Studies of a Bacillus-conserved Nucleotide Pyrophosphorylase, BcMazG
Published on: May 16, 2017
10:13Production and Visualization of Bacterial Spheroplasts and Protoplasts to Characterize Antimicrobial Peptide Localization
Published on: August 11, 2018
Related Concept Videos
01:09Recombinant DNA
01:23Antimicrobial Proteins
Interferons
Interferons (IFNs) are proteins produced by lymphocytes, macrophages, and fibroblasts infected with viruses. While IFNs cannot prevent viruses from entering and...