Cost-effective fibrinolytic enzyme production by microalga Dunaliella tertiolecta using medium supplemented with corn steep liquor.

A fibrinolytic enzyme from the microalga Dunaliella tertiolecta was produced under mixotrophic conditions using different corn steep liquor (CSL) concentrations ( 0 ≤ CLS ≤ 0.75%), purified using a combination of salting out and ion-exchange chromatography, and then biochemical characterized. Cultivation of this microalga using 0.5% CSL led to the highest maximum cell concentration (1.960±0.010 mg L-1) and cell productivity (0.140g L-1 day-1), besides a high fibrinolytic activity of the extract obtained by the homogenization method (102 ±1 U mL-1). The enzyme extracted from the microalgal biomass was 5-fold purified with a 20% yield and was found to have a specific activity of 670 U mg-1. The enzyme, whose molecular weight determined by fibrin zymography was 10 kDa, was shown to be stable at pH 3.0-9.0 and up to 70°C with optimal pH and temperature values of 8.0 and 50°C, respectively. When compared to other fibrinolytic enzymes, this protease stood out for its high fibrinolytic activity, which was enhanced by Fe2+, inhibited by Zn2+, Cu2+, Mg2+, and Ca2+, and strongly inhibited by phenylmethylsulfonyl fluoride, suggesting that it belongs to the serine metalloprotease family. Moreover, thanks to its thermal stability, the enzyme may be easily preserved and activated under high-temperature conditions.

Comparative study of structures and functional motifs in lectins from the commercially important photosynthetic microorganisms.

Photosynthetic microorganisms, specifically cyanobacteria and microalgae, can synthesize a vast array of biologically active molecules, such as lectins, that have great potential for various biotechnological and biomedical applications. However, since the structures of these proteins are not well established, likely due to the presence of intrinsically disordered regions, our ability to better understand their functionality is hampered. We embarked on a study of the carbohydrate recognition domain (CRD), intrinsically disordered regions (IDRs), amino acidic composition, as well as and functional motifs in lectins from cyanobacteria of the genus Arthrospira and microalgae Chlorella and Dunaliella genus using a combination of bioinformatics techniques. This search revealed the presence of five distinctive CRD types differently distributed between the genera. Most CRDs displayed a group-specific distribution, except to C. sorokiniana possessing distinctive CRD probably due to its specific lifestyle. We also found that all CRDs contain short IDRs. Bacterial lectin of Arthrospira prokarionte showed lower intrinsic disorder and proline content when compared to the lectins from the eukaryotic microalgae (Chlorella and Dunaliella). Among the important functions predicted in all lectins were several specific motifs, which directly interacts with proteins involved in the cell-cycle control and which may be used for pharmaceutical purposes. Since the aforementioned properties of each type of lectin were investigated in silico, they need experimental confirmation. The results of our study provide an overview of the distribution of CRD, IDRs, and functional motifs within lectin from the commercially important microalgae.

Algae as a source of peptides inhibitors of the angiotensin-converting enzyme: a systematic review.

Hypertension is a factor that contributes to the risk of chronic diseases. The inhibition of angiotensin-I converting enzyme (ACE) is a useful therapeutic approach to the hypertension treatment. The algae have been an alternative for the production of ACE inhibitory (ACEi) peptides from enzymatic hydrolysis due to their protein-rich biomass. The aim of this study was to systematically review the literature regarding the production, composition and activity of ACEi peptides derived from algae proteins. Systematic database searches identified 648 related articles. Among these, only 14 were selected according to the eligibility criteria to this review. Macroalgae are more studied than microalgae as sources of ACEi peptides. Furthermore, hydrolysates by thermolysin or bromelain exhibited the highest ACEi activity compared to other enzymes. The main features of the peptides with high ACE inhibition are low molecular weight, short amino acids sequence and non-competitive inhibition pattern. In vivo studies using hydrolysates and peptides derived from algae proteins showed antihypertensive activity in spontaneously hypertensive rats (SHR). Thus, it is suggested that ACEi peptides derived from algae can be considered as potential antihypertensive.

Bioactive Compounds of Arthrospira spp. (Spirulina) with Potential Anticancer Activities: A Systematic Review.

Arthrospira, a genus of blue-green cyanobacteria, is known for its great biological activity due to the presence of a large number of substances that are potentially active against tumor cells. This review aimed to evaluate the potential of Arthrospira spp. for the treatment or reduction of several types of cancer, in addition to elucidating the mechanism of action by which their compounds act on tumor cells. A systematic review was carried out in PubMed, Science Direct, LILACS, and SciELO databases, including original studies from 2009 to 2020. A total of 1306 articles were independently assessed according to the eligibility criteria, of which 20 articles were selected and assessed for the risk of bias using seven criteria developed by the authors. Arthrospira spp. of cyanobacteria have been evaluated against eight different types of cancer, mainly colon cancer. Among all the compounds, phycocyanin was the most used, followed by peptides and photosensitizers. In general, compounds from Arthrospira spp. act as anticancer agents by inhibiting the proliferation of tumor cells, triggering cell cycle arrest, and inducing apoptosis via different signaling pathways. In addition, these compounds also exhibited antioxidant, antiangiogenic, and antimetastatic activities. Phycocyanin demonstrated better efficacy against several types of cancer via different activities and therapeutic targets. Furthermore, it was the only molecule that functioned in synergy with other drugs that are already well established for the treatment of cancer.

Looking for alternative treatments for bovine and caprine mastitis: Evaluation of the potential of Calliandra surinamensis leaf pinnulae lectin (CasuL), both alone and in combination with antibiotics.

This work aimed to evaluate the effects of CasuL on growth and viability of 15 mastitis isolates from cows and goats, to determine the synergistic potential between CasuL and antibiotics, and to investigate the effects on bacterial ultrastructure and antibiofilm activity. The lectin inhibited the growth of Staphylococcus isolates from either bovine (Ssp6PD and Sa) or caprine (Ssp5D and Ssp01) mastitis. The minimal inhibitory concentrations were ranged from 3.75 to 15 µg/ml. Synergistic effect was observed for CasuL-tetracycline against Sa and Ssp6PD and CasuL-ampicillin against Ssp01. No structural damage was observed under the scanning electron microscope in CasuL treatments. Flow cytometry analysis using thiazol orange and propidium iodide demonstrated that CasuL was unable to reduce the cell viability of the isolates tested. At sub-inhibitory concentrations, CasuL reduced biofilm formation by the isolates Sa and Ssp5D. However, CasuL-tetracycline and CasuL-ampicillin combinations inhibited biofilm formation by Ssp6PD and Ssp01, respectively. In conclusion, CasuL is a bacteriostatic and antibiofilm agent against some mastitis isolates and displayed a synergistic potential when used in combination with either ampicillin (against one isolate) or tetracycline (against two isolates). The results stimulate the evaluation of CasuL for the treatment of mastitis, particularly when used in conjunction with antibiotics.

Podem as bactérias ácido lácticas probióticas apresentarem efeito antitumoral em modelo animal de câncer de cólon? Uma revisão da literatura / Can probiotic acid lactic bacteria submit antitumor effect in animal model of colon cancer? A review of the literature

O Câncer de cólon é um dos tipos mais comuns de câncer no mundo e a segunda principal causa de morte relacionada a esta doença em países desenvolvidos. Até 75% dos casos estão associados com a alimentação, indicando que uma pessoa pode reduzir o seu risco simplesmente através de modificação na dieta. Estudos em animais demonstram que várias cepas de bactérias ácido lácticas protegem contra o câncer de cólon em roedores, embora os dados em humanos sejam limitados e conflitantes. O objetivo deste estudo foi investigar a eficácia das bactérias ácido lácticas no tratamento e redução do câncer de cólon em modelo animal. Foram realizadas buscas sistemáticas em bases de dados eletrônicas alcançando 1079 artigos relacionados, entretanto apenas 6 artigos foram eleitos de acordo com os critérios de elegibilidade para análise. Todos os artigos avaliados apresentaram resultados satisfatórios quanto à inibição do câncer de cólon em ratos e camundongos ao utilizarem cepas predominantemente do gênero Lactobacillus. Este estudo pode responder a hipótese de que as bactérias ácido lácticas apresentam efeito preventivo e antitumoral contra o câncer de cólon.(AU)

Colon cancer is one of the most common types of cancer in the world and the second leading cause of death related to the disease in developed countries. Up to 75% of cases are associated with eating, indicating that a person can reduce their risk simply through dietary modification. Studies in animals show that various strains of lactic acid bacteria protect against colon cancer in rodents although data in humans are limited and conflicting. The aim of this study was to investigate the efficacy of acid lactic bacteria in the treatment and reduction of colon cancer in animal models. Systematic searches were conducted in electronic databases reaching 1079 related articles, only six articles were elected according instead of to the eligibility criteria for analysis. All reviewed articles showed satisfactory results on the inhibition of colon cancer in rats and mice when using predominantly Lactobacillus strains. This study can answer hypothesis that acid lactic bacteria has antitumor effect against colon cancer.(AU)

Fibrinolytic protease production by new Streptomyces sp: DPUA 1576 from Amazon lichens

Background Streptomyces sp. DPUA 1576 from Amazon lichens was studied to protease and fibrinolytic production. A 2² factorial experimental design was applied to optimize its protease enzyme production using two independent variables, namely soybean flour and glucose concentrations. Results The optimal conditions to obtain high protease production (83.42 U/mL) were 1.26% soybean flour and 1.23% glucose concentration. A polynomial model was fitted to correlate the relationship between the two variables and protease activity. In relation to fibrinolytic activity, the highest activity of 706.5 mm² was obtained at 1.7% soybean flour and 1.0% glucose concentration, which was 33% higher than plasmin. Fibrinolytic production was not optimized in the studied conditions. Conclusions These results show that the optimization of the culture medium can enhance protease production, thus becoming a good process for further research. In addition, Streptomyces sp. DPUA 1576, isolated from Amazon lichens, might be a potential strain for fibrinolytic protease production.

Ethanol effect on batch and fed-batch Arthrospira platensis growth.

The ability of Arthrospira platensis to use ethanol as a carbon and energy source was investigated by batch process and fed-batch process. A. platensis was cultivated under the effect of a single addition (batch process) and a daily pulse feeding (fed-batch process) of pure ethanol, at different concentrations, to evaluate cell concentration (X) and specific growth rate (µ). A marked increase was observed in the cell concentration of A. platensis in runs with ethanol addition when compared to control cultures without ethanol addition. The fed-batch process using an ethanol concentration of 38 mg L(-1) days(-1) reached the maximum cell concentration of 2,393 ± 241 mg L(-1), about 1.5-fold that obtained in the control culture. In all experiments, the maximum specific growth rate was observed in the early exponential phase of cell growth. In the fed-batch process, µ decreased more slowly than in the batch process and control culture, resulting in the highest final cell concentration. Ethanol can be used as a feasible carbon and energy source for A. platensis growth via a fed-batch process.

CO2 from alcoholic fermentation for continuous cultivation of Arthrospira (Spirulina) platensis in tubular photobioreactor using urea as nitrogen source.

Carbon dioxide released from alcoholic fermentation accounts for 33% of the whole CO(2) involved in the use of ethanol as fuel derived from glucose. As Arthrospira platensis can uptake this greenhouse gas, this study evaluates the use of the CO(2) released from alcoholic fermentation for the production of Arthrospira platensis. For this purpose, this cyanobacterium was cultivated in continuous process using urea as nitrogen source, either using CO(2) from alcoholic fermentation, without any treatment, or using pure CO(2) from cylinder. The experiments were carried out at 120 µmol photons m(-2) s(-1) in tubular photobioreactor at different dilution rates (0.2 ≤ D ≤ 0.8 d(-1) ). Using CO(2) from alcoholic fermentation, maximum steady-state cell concentration (2661 ± 71 mg L(-1) ) was achieved at D = 0.2 d(-1) , whereas higher dilution rate (0.6 d(-1) ) was needed to maximize cell productivity (839 mg L(-1) d(-1) ). This value was 10% lower than the one obtained with pure CO(2) , and there was no significant difference in the biomass protein content. With D = 0.8 d(-1) , it was possible to obtain 56% ± 1.5% and 50% ± 1.2% of protein in the dry biomass, using pure CO(2) and CO(2) from alcoholic fermentation, respectively. These results demonstrate that the use of such cost free CO(2) from alcoholic fermentation as carbon source, associated with low cost nitrogen source, may be a promising way to reduce costs of continuous cultivation of photosynthetic microorganisms, contributing at the same time to mitigate the greenhouse effect.