A new milk-clotting enzyme from noni seeds (Morinda citrifolia L) / Uma nova enzima coagulante de leite de sementes de noni (Morinda citrifolia L.)

They are found in all living organisms and in plants has received more attention because of their potential involvement in various industrial processes. Noni plant (Morinda citrifolia L.), belongs to the family Rubiaceae, of southwestern Asia origin. The juice made with the fruit is widely used as phytotherapic agent and in combating a range of diseases, while seeds are usually discarded. The objective of this work was to seek active milk clotting proteases in noni seeds. The crude extract (CE) protein was obtained from seeds sprayed and mixed with tris-HCl 50mM buffer (pH 6.0). The results demonstrated the presence of proteases with milk clotting properties in noni seeds, which a high milk clotting activity from 3,891 U/mL to 65°C.  This value indicates that 1mL of extract is able to coagulate 3.8 liters of milk in 40 minutes under specific conditions. This property indicates the potential of noni seeds as a natural source of proteolytic enzymes with biotechnological potential for application in the cheese making industry.

As proteases são um grupo de enzimas que catalisam várias reações essenciais. Eles são encontrados em todos os organismos vivos e nas plantas, tem recebido mais atenção devido ao seu potencial envolvimento em vários processos industriais. A planta noni (Morinda citrifolia L.), pertence à família Rubiaceae, de origem no sudoeste da Ásia. O suco feito com a fruta é amplamente utilizado como agente fitoterápico e no combate a uma série de doenças, enquanto as sementes costumam ser descartadas. O objetivo deste trabalho foi buscar proteases ativas da coagulação do leite em sementes de noni. O extrato bruto proteico (CE) foi obtido de sementes pulverizadas e misturadas com tampão tris-HCl 50mM (pH 6,0). Os resultados demonstraram a presença de proteases com propriedades de coagulação do leite em sementes de noni, que apresentam alta atividade de coagulação do leite de 3.891 U / mL a 65 ° C. Esse valor indica que 1mL de extrato é capaz de coagular 3,8 litros de leite em 40 minutos em condições específicas. Esta propriedade indica o potencial das sementes de noni como fonte natural de enzimas proteolíticas com potencial biotecnológico para aplicação na indústria de produção de queijo.

Biochemical characterization and phytotoxic activity of protein extract from Euphorbia tirucalli L.

ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia tirucalli L., a tropical and subtropical plant, also known by the popular name avelós, has been used in folk medicine against many diseases as rheumatism, asthma, toothache, and cancer. Studies have shown that natural compounds contained in this plant species may be associated with these functions. However, little is known about its potential toxicity. AIM OF THE STUDY: Several proteins conduct biological functions, in particular, proteinases, play a crucial role in many mechanisms of living beings, including plants, animals and microorganisms. However, when poorly regulated, they can generate consequences, such as the non-production of certain substances, or even the abnormal multiplication of cells, which leads to tumors. On the other hand, by regulating these enzymes, proteinase inhibitors act by reducing the activity of proteinases, thus preventing their malfunction. The objective of this work was to evaluate the toxicity of the protein extract of E. tirucalli and to purify a protease inhibitor that may be associated with the biological medicinal functions of the plant. MATERIALS AND METHODS: The cytotoxic and mutagenic properties of the protein extract produced from the stem of avelós was investigated using the Ames test. The protein extract was also submitted to a protease inhibitor purification process using the gel filtration chromatography technique and the purified protein was biochemically characterized. RESULTS: A protease inhibitor, called tirustatin, was isolated 1.84-fold by Biogel P100. The calculated molecular mass of the isolated protein is 25.97 kDa. The inhibitor was stable at pH 3-10, with pronounced activity at pH 6. Thermostability was observed even at elevated temperature (100 °C) with inhibitory activity increased by 1.14-fold compared to inhibitor activity at room temperature. Incubation at basic pH values for up to 60 min caused little reduction (0.25-fold) in the papain inhibitory activity of tirustatin. The stoichiometry of the papain-tirustatin interaction was 1.5: 1 and 28.8 pM of the inhibitor effected 50% inhibition. With an equilibrium dissociation constant of 8.74 x 10-8M for the papain enzyme, it is possible to evaluate the isolated protein as a non-competitive inhibitor. In addition, the protein extract of E. tirucalli even at the maximum concentration used (20 µg/mL), did not show a cytotoxic and mutagenic profile in a bacterial model. CONCLUSION: The results presented in this work provide data that reinforce the idea of the potential use of proteins produced in E. tirucalli as pharmacological and biotechnological agents that can be exploited for the development of efficient drugs.

In vitro biochemical characterization and genotoxicity assessment of Sapindus saponaria seed extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Sapindus saponaria, also popularly known as soapberry, has been used in folk medicinal values because of its therapeutic properties and several compounds in its composition, which represent a target in potential for drug discovery. However, few data about its potential toxicity has been reported. AIM OF THE STUDY: Plant proteins can perform essential roles in survival, acting as defense mechanism, as well functioning as important molecular reserves for its natural metabolism. The aim of the current study was to investigate the in vitro toxicity profile of protein extract of S. saponaria and detect protein potentially involved in biological effects such as collagen hydrolysis and inhibition of viral proteases. MATERIALS AND METHODS: Protein extract of soapberry seeds was investigated for its cytotoxic and genotoxic action using the Ames test. The protein extract was also subjected to a partial purification process of a protease and a protease inhibitor by gel chromatography filtration techniques and the partially isolated proteins were characterized biochemically. RESULTS: Seed proteins extract of S. saponaria was evaluated until 100 µg/mL concentration, presenting cytotoxicity and mutagenicity in bacterial model mostly when exposed to exogenous metabolic system and causing cytotoxic and genotoxic effects in HepG2 cells. The purification and partial characterization of a serine protease (43 kDa) and a cysteine protease inhibitor (32.8 kDa) from protein extract of S. Saponaria, corroborate the idea of ​​the biological use of the plant as an insecticide and larvicide. Although it shows cytotoxic, mutagenic and genotoxic effects. CONCLUSION: The overall results of the present study provide supportive data on the potential use of proteins produced in S. saponaria seeds as pharmacological and biotechnological agents that can be further explored for the development of new drugs.

Control of papaya fruits anthracnose by essential oil of Ricinus communis

The aim of this work was to investigate the potential of castor oil for the control of papaya diseases caused by the fungus Colletotrichum gloeosporioides and the bacterium Pseudomonas caricapapayae. The treatment with 1% castor oil did not significantly affect the fungal growth. The effectiveness of castor oil for the control of anthracnose was shown when 5% and 10% (v/v) were used in the assays resulting in reduced mycelial growth. Fungal sporulation was strongly inhibited at 10% (v/v) concentration of essential oil. The studies with the fresh fruits treated with 5% (v/v) castor oil in aqueous emulsions resulted in effective reduction of pathogen spread in these fruits. No lesion was found in the fruits treated with oil, when compared to the control fruits. Castor oil showed no effect against the P. caricapapayae when tested in vitro. These results suggested the potential use of the castor bean essential oil and its fatty acids constituents for the control of anthracnose in papaya fruits.