Purification and stability analysis of antimicrobial proteins from Varuna litterata. (April 2021)

• Record Type: Journal Article
• Authors: Wang, Liming; Sathuvan, Malairaj; Chen, Shengqin; Cai, Kun; Chen, Shan; Liu, Yang
• Is Part Of: Microbial pathogenesis. Volume 153(2021)
• Publication Date: 2021-04
• Subjects: Marine species -- Varuna litterata -- Antimicrobial protein -- Hemocyanin -- Phenol oxidase activity; Hemocyanin HMC -- Staphylococcus aureus S, aureus -- Escherichia coli E coli -- ammonium sulfate AS -- positive control PC -- negative control NC -- flat crab antimicrobial protein FCAP -- flat crab antimicrobial protein crude extract FCAP1 -- flat crab antimicrobial protein salting-out and redissolved solution FCAP2 -- flat crab antimicrobial protein gel column chromatography purification solution FCAP3 -- flat crab antimicrobial protein ion column chromatography water elution peak FCAP4 -- flat crab antimicrobial protein ion column chromatography salt elution peak FCAP5 -- Antimicrobial protein-peptide of flat crab FCAP6 -- polyacrylamide gel electrophoresis PAGE -- levodopa l-DOPA -- mass spectra MS -- liquid chromatogram LC -- Dalton Da -- sodium dodecyl sulfate SDS -- rate of inhibition RI -- Antimicrobial rate ABR; Pathogenic microorganisms -- Periodicals; Pathology, Molecular -- Periodicals; Communicable Diseases -- microbiology -- Periodicals; Communicable Diseases -- parasitology -- Periodicals; Micro-organismes pathogènes -- Périodiques; Pathologie moléculaire -- Périodiques; Electronic journals; 616.9041
• DOI: 10.1016/j.micpath.2020.104637
• Languages: English
• ISSNs: 0882-4010

Abstract

Abstract: Ten marine species, including different crabs, bivalve molluscs, and fish intestines were selected to screen the natural antimicrobial protein or peptide as they are enriched with various microorganisms. The crude extract from Varuna litterata, a marine crab which is used as a raw material in the preparation of pickled crabs in Chaoshan area of China, was proved to have a potent bacteriostatic effect against gram-negative bacterium ( Escherichia coli ) and gram-positive bacterium( Staphylococcus aureus ) compared with other marine species. The crude proteins of Varunalitterata were salted-out for preliminary purification and further purified by gel filtration (Sephadex G-150) or anion exchange (DEAE-cellulose 52) chromatographic column. An increase in the antimicrobial activity was noted with the increase in the purity level of the protein. A relatively pure protein was eventually obtained, which was determined to be belonging to the hemocyanin family based on the mass spectrometric data analysis. The purified proteins solution (1 mg/ml) from Varuna litterata exhibited similar antimicrobial activity to that of gentamycin sulfate (0.2 mg/ml), which were relatively stable in a certain pH or temperature range. A structure-activity relationship of the purified hemocyanin was determined based on the interaction of hemocyanin and different chromatographic medium, which revealed that the integrated hexamers played a remarkable role in its bacteriostatic activity. Moreover, the phenoloxidase activity of hemocyanin from Varuna litterata was found as the underlying cause of its antimicrobial potential. Highlights: Proteins extracted from Varuna litterata has evident bacteriostasis. Hemocyanin was proved to have a strong bacteriostatic effect in Varuna litterata. Hexamers in hemocyanin play an important role in bacteriostasis.