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Fish Shellfish Immunol. 2016 Apr;51:9-16. doi: 10.1016/j.fsi.2016.02.004. Epub 2016 Feb 12.

A preliminary study on the antibacterial mechanism of Tegillarca granosa hemoglobin by derived peptides and peroxidase activity.

Author information

1
Zhejiang Key Laboratory of Aquatic Germplasm Resources, College of Biological & Environmental Sciences, Zhejiang Wanli University, Zhejiang 315100, PR China.
2
Zhejiang Key Laboratory of Aquatic Germplasm Resources, College of Biological & Environmental Sciences, Zhejiang Wanli University, Zhejiang 315100, PR China; School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, PR China.
3
School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, PR China.
4
Zhejiang Key Laboratory of Aquatic Germplasm Resources, College of Biological & Environmental Sciences, Zhejiang Wanli University, Zhejiang 315100, PR China. Electronic address: sufang@gmail.com.
5
Zhejiang Key Laboratory of Aquatic Germplasm Resources, College of Biological & Environmental Sciences, Zhejiang Wanli University, Zhejiang 315100, PR China. Electronic address: zhihua9988@126.com.

Abstract

The blood clam, Tegillarca granosa, is one of the few bivalve molluscs containing hemoglobin (Hb). In the present study, we purified two types of T. granosa hemoglobin, Tg-HbI and Tg-HbII, using size exclusion chromatography and measured their antibacterial and peroxidase activities. We also tested antibacterial activities of peptides prepared by trypsin digestion of purified Tg-Hb and reversed-phase high-performance liquid chromatography purification. Purified Tg-HbI and Tg-HbII showed antibacterial activity against Escherichia coli, Pseudomonas putida, Bacillus subtilis, and Bacillus firmus, with differences in minimal inhibitory concentrations (MICs), but lacked antibacterial activity against Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio harveyi and Staphylococcus aureus. In contrast, 7 Tg-Hb derived peptides exhibited varying degrees of antibacterial activity against V. alginolyticus (MICs: 12-200 μg/ml), V. parahaemolyticus (11-100 μg/ml) and V. harveyi (1-200 μg/ml). The antibacterial activity of Hb derived peptides was confirmed by fluorescence microscopy. In addition, peroxidase activity was detected in Tg-HbI and Tg-HbII. The results indicated that in addition to functioning as a respiratory protein T. granosa hemoglobins likely play a role in host antibacterial defense probably via a peroxidase activity of native molecules and some internal peptides released from the proteins.

KEYWORDS:

Antibacterial activity; Hemoglobin; MIC; Peptides; Peroxidase activity; Tegillarca granosa

PMID:
26876330
DOI:
10.1016/j.fsi.2016.02.004
[Indexed for MEDLINE]

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