A novel lectin from mushroom Phellodon melaleucus displays hemagglutination activity,and antitumor activity in a B16 melanoma mouse model

A novel lectin(termed PML)was purified from fruiting bodies of the edible mushroom Phellodon melaleucus(division Basidiomycota)by ion exchange,hydrophobic interaction,and gel filtration chromatographies,with overall titer recovery~60%and 20-fold purification.PML displayed hemagglutination activity 13319 units/mg toward rabbit erythrocytes.SDS-PAGE and gel filtration analyses revealed that PML is a homodimeric lectin with a molecular weight of 28.8 kDa.PML hemagglutination activity was not inhibited by various simple sugars or their derivatives,but was enhanced by cations Ca^(2+),Mg^(2+),Zn^(2+),and Cu^(2+).The activity was stable in pH range 6–9 and in the temperature range 20–60°C.Circular dichroism(CD)spectroscopic analysis showed that PML was composed primarily ofβ-sheets with lowα-helix content.In a B16 melanoma mouse model,PML treatment significantly inhibited tumor growth,and increased cytokine IL-10 content.Our findings suggest that PML is a potential anticancer therapeutic agent.

Geographically Distinct Expression Profile of Host Defense Peptides in the Skin of the Chinese Odorous Frog, Odorrana margaretae

Odorrana margaretae （Anura： Ranidae） is widely distributed in the southern provinces of China. Previously, 72 antimicrobial peptides （AMPs） belonging to 21 families were identified from the skin of O. margaretae, which were captured in the Hunan province. In the present study, five O. margaretae frogs were captured from the Guizhou province and a total of 28 cDNAs encoding 17 host defense peptides （HDPs） belonging to 14 families were cloned from the skin cDNA library of O. margaretae. Among the 17 HDPs, only one （brevinin-1-Omar5） had been characterized. The distinct HDP expression profiles for O. margaretae in the previous and present study may be attributed to the environmental differences between the sampling locations and the genetic divergence among O. margaretae populations. Besides, 11 of the 17 HDPs identified in the present study were novel for ranids. In order to understand their roles in host defense reactions, three HDPs （odorranain-H-OM1, odorranain-M-OM and ranatuerin-2-OM）, which possess low sequence similarity with the known amphibian HDPs, were selected for further chemical synthesis and functional analysis. Odorranain-H-OM1 showed direct antimicrobial activity against bacteria and fungi. Odorranain-M-OM exhibited concentration-dependent anti-oxidant activity. Ranatuerin-2-OM showed lectin-like activity and could strongly hemagglu -tinate human intact erythrocytes with or without the presence of Ca2＋. The diverse activities of HDPs implied that they may play different roles in host defense reactions of O. margaretae.

In Vitro Evaluation of Gamma Irradiation on a Gel Formulation of Cratylia Mollis: Rheological Proporties and Microbiological Control

Lectin Cramoll-1,4, obtained from Cratylia mollis seeds (beans camaratu) was structurally characterized, biologically and pharmacologically, but its use as a biopharmaceutical is not well documented. The objective of this study is to propose a biopharmaceutical formulation lectin Cramoll-1,4, test their hemagglutinating properties in vitro as well as the use of gamma radiation as a continuous process of decontamination formulation. It was made of the extraction and purification Cramoll-1,4, was developed a gel formulation using Carbopol? as a vehicle, at concentrations of 50, 100, 200, 300 and 600 μg was irradiated with 60Co gamma rays in a dose of 7.549 kGy·h–1. The proposed formulation at a concentration of 300 μg produced an increase in the hemagglutinating units Cramoll-1,4 due to the synergistic effect caused by gamma radiation. Considering the diverse use of lectins, specific molecular and structural factors, as well as changes resulting from its formulation, concentration, irradiation and route of administration is of utmost importance to continue the studies in vitro, for subsequent application in vivo to characterize the physiological and molecular processes involved in the response and cellular effects.