Antifungal activity of Aplysianin E, A cytotoxic protein of sea hare (Aplysia kurodai) eggs

Lijima, Ryosuke; Kisugi, Jun; Yamazaki, Masatoshi

doi:10.1016/0145-305x(94)00055-k

Cited by 36 publications

(17 citation statements)

References 25 publications

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“…However, there are implications for how escapin, as the dominant protein in the defensive ink secretion of Aplysia californica, may also function as an antipredatory or antiseptic compound. L-amino acid oxidase (LAAO) activity of antimicrobial and antineoplastic agents from a variety of species has been reported as a major (Iijima et al, 1995;Suhr and Kim, 1996;Torii et al, 1997;Ehara et al, 2002;Lu et al, 2002;Wei et al, 2003;Butzke et al, 2004) or partial (Jimbo et al, 2003;Kanzawa et al, 2004) mechanism for their effects. Escapin also possesses LAAO activity: it shows strong and rapid activity under our assay conditions when using arginine or lysine as a substrate, with reactions being completed within 30·s at room temperature (Fig.·4).…”

Section: Discussionmentioning

confidence: 99%

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Yang

Johnson

et al. 2005

Journal of Experimental Biology

108

122

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SUMMARY A 60 kDa monomeric protein isolated from the defensive purple ink secretion of the sea hare Aplysia californica was cloned and sequenced, and is the first sea hare antimicrobial protein to be functionally expressed in E. coli. Sequence analysis suggested that this protein is a flavin-containing l-amino acid oxidase (LAAO), with one predicted potential glycosylation site, although the glycosylation could not be experimentally confirmed. This protein, which we call `escapin', has high sequence similarity to several other gastropod proteins. Escapin was verified by NMR, mass spectroscopy and HPLC to have FAD as its flavin cofactor. Escapin's antimicrobial effects, bacteriostasis and bactericidal, were determined using a combination of two assays: (1) incubation of bacteria on solid media followed by assessment of inhibition by direct observation of zones of inhibition or by turbidity measurements; and (2) incubation of bacteria in liquid media followed by counting viable colonies after growing on agar plates. Native escapin inhibited the growth of Gram-positive and Gram-negative bacteria, including marine bacteria (Vibrio harveyiiand Staphylococcus aureus) and pathogenic bacteria(Staphylococcus aureus, Streptococcus pyogenes and Pseudomonas aeruginosa). Escapin also inhibited the growth of yeast and fungi, with different efficacies. Escapin's antimicrobial activity was concentration dependent and did not decrease when stored for more than 5 months at room temperature. Escapin was bacteriostatic and not bactericidal in minimal media (e.g. salt media) with glucose, yeast extract, and a mixture of 20 amino acids each at 50 μmol l-1, but was bactericidal in media enriched with Tryptone Peptone. Escapin was also strongly bactericidal in media with l-lysine at concentrations as low as 3 mmol l-1 and slightly bactericidal in 50 mmol l-1l-arginine, but not in most other amino acids even at 50 mmol l-1. Escapin had high oxidase activity (producing hydrogen peroxide) with either l-arginine or l-lysine as a substrate and little to no oxidase activity with other l-amino acids. Hydrogen peroxide alone (without escapin or amino acids) was strongly bacteriostatic but poorly bactericidal, similar in this respect to l-arginine but different from l-lysine in the presence of escapin. Together these results suggest that there are multiple mechanisms to escapin's antimicrobial effects, with bacteriostasis resulting largely or entirely from the effects of hydrogen peroxide produced by escapin's LAAO activity, but bactericidal effects resulting from lysine-dependent mechanisms not directly involving hydrogen peroxide. Recombinant escapin expressed in bacteria was also active against Gram-positive and Gram-negative bacteria,suggesting that glycosylation is not essential for antimicrobial activity.

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Section: Discussionmentioning

confidence: 99%

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Yang

Johnson

et al. 2005

Journal of Experimental Biology

108

122

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“…Beckwith et al (8) reported that the small peptides, dolastatins, isolated from Dollabella auricularia, demonstrate an antineoplastic effect and induce apoptosis in some human lymphoma cell lines. Several researchers (9)(10)(11)(12)(13)(14) have isolated aplysianin A, E and P from the albumen gland, the egg and the purple fluid, respectively, of Aplysia kurodai. Aplysianins are high molecular weight glycoproteins which have antibacterial and antineoplastic effects the same as dolastatins and inhibit the synthesis of deoxyribonucleic acid and ribonucleic acid, just as an effect of a chemotherapeutic drug.…”

Section: Discussionmentioning

confidence: 99%

Acute Liver Damage with Characteristic Apoptotic Hepatocytes by Ingestion of Aplysia kurodai, a Sea Hare.

et al. 1998

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A case of acute liver damage by ingestion ofAplysia kurodai, a sea hare is reported. A 40-yearold man, complaining of vomiting and pyrexia after eating a sea hare, was admitted. Laboratory data showed mild liver damage with sustained elevations of aminotransferases. Microscopic findings in the liver biopsy specimen revealed characteristic apoptotic hepatocytes accompanied by mitotic hepatocytes. It is suggested that bioactive substances in the sea hare might induce such apoptosis of hepatocytes in the liver. (Internal Medicine 37: 927-929, 1998)

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“…This is bacteriostatic against various Gram-positive and Gram-negative bacteria at 0.2–5.8 μg mL −1 and seems to function by completely inhibiting the syntheses of bacterial DNA and RNA; a property that also lends it cytotoxic activity against eukaryotic tumour cells [184]. Related aplysianins are present in the albumin gland (aplysianin A) and eggs (aplysianin E) of A. kurodai [221,222]. The latter must certainly help the eggs from being colonized by bacterial and fungal epibionts.…”

Section: Unconventional Anti-infectivesmentioning

confidence: 99%

Conventional and Unconventional Antimicrobials from Fish, Marine Invertebrates and Micro-algae

2010

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All eukaryotic organisms, single-celled or multi-cellular, produce a diverse array of natural anti-infective agents that, in addition to conventional antimicrobial peptides, also include proteins and other molecules often not regarded as part of the innate defences. Examples range from histones, fatty acids, and other structural components of cells to pigments and regulatory proteins. These probably represent very ancient defence factors that have been re-used in new ways during evolution. This review discusses the nature, biological role in host protection and potential biotechnological uses of some of these compounds, focusing on those from fish, marine invertebrates and marine micro-algae.

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Antifungal activity of Aplysianin E, A cytotoxic protein of sea hare (Aplysia kurodai) eggs

Cited by 36 publications

References 25 publications

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Acute Liver Damage with Characteristic Apoptotic Hepatocytes by Ingestion of Aplysia kurodai, a Sea Hare.

Conventional and Unconventional Antimicrobials from Fish, Marine Invertebrates and Micro-algae

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