Characterization, functional analysis and antibacterial activities of theromacin from the Akoya pearl oyster Pinctada fucata

Date
2021
Authors
Guo, Hua-Yang
Liu, Bo
Yang, Jing-Wen
Jiang, Shi-Gui
Zhang, Dian-Chang
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73
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Abstract
The Akoya Pearl oyster Pinctada fucata is distributed over the coastal area of South China and is the most popular farming shellfish for seawater pearl production. Antimicrobial peptides (AMPs) could inhibit or kill pathogenic microorganisms. Theromacin has been proven to be an antimicrobial peptide, which plays an essential role in the body's immune system. The AMP gene to be identified was encoding theromacin in the pearl oyster Pinctada fucata (PoAP). The full-length PoAP cDNA contains 522 bp and consists of a 5’-UTR of 6 bp, an unusually long 3’-UTR of 749 bp, and an open reading frame (ORF) of 375 bp that encodes a 124 amino acid peptide with a molecular weight of 13.67 kDa and the theoretical isoelectric point of 9.25. Homology analysis of the deduced amino acid sequence of the PoAP with other known theromacin sequences by MatGAT software revealed that the PoAP shared 29.0%-46.8% similarity to the other known theromacin sequences. Signal P-N program showed that PoAP contains 33 amino acid signal peptides and a mature peptide located at amino acids 34-124; the mature peptide contains 12 cysteine residues and 13 alkaline amino acid residues with a molecular weight of 13.67 kDa and the theoretical isoelectric point of 9.25. The tempered program suggests that PoAP is a membrane protein with one transmembrane helix between amino acids 13 and 34. The PoAP gene was linked into prokaryotic vector pET-32α, and the PoAP fusion protein with 31 kDa molecular mass was successfully expressed in Escherichia coli BL21. Using His-Bind Purification Kit Protocol purifies the antimicrobial peptides recombinant protein and compares the bacteriolysis effect on Gram-positive and Gram-negative bacteria using the purified protein. The results show that PoAP proteins had a bacteriolysis effect on the Gram-negative bacteria but on Gram-positive organisms. The antibacterial peptides theromacin seem to play a crucial antibacterial function in the immune responses of pearl oysters.
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Pinctada fucata, pearl oyster, antimicrobial peptides, theromacin, antibacterial activities
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11 pages
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The Israeli Journal of Aquaculture - Bamidgeh
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