Oligosaccharides production from coprophilous fungi: An emerging functional food with potential health-promoting properties.

Functional foods are essential food products that possess health-promoting properties for the treatment of infectious diseases. In addition, they provide energy and nutrients, which are required for growth and survival. They occur as prebiotics or dietary supplements, including oligosaccharides, processed foods, and herbal products. However, oligosaccharides are more efficiently recognized and utilized, as they play a fundamental role as functional ingredients with great potential to improve health in comparison to other dietary supplements. They are low molecular weight carbohydrates with a low degree of polymerization. They occur as fructooligosaccharide (FOS), inulooligosaccharadie (IOS), and xylooligosaccahride (XOS), depending on their monosaccharide units. Oligosaccharides are produced by acid or chemical hydrolysis. However, this technique is liable to several drawbacks, including inulin precipitation, high processing temperature, low yields, and high production costs. As a consequence, the application of microbial enzymes for oligosaccharide production is recognized as a promising strategy. Microbial enzymatic production of FOS and IOS occurs by submerged or solid-state fermentation in the presence of suitable substrates (sucrose, inulin) and catalyzed by fructosyltransferases and inulinases. Incorporation of FOS and IOS enriches the rheological and physiological characteristics of foods. They are used as low cariogenic sugar substitutes, suitable for diabetics, and as prebiotics, probiotics and nutraceutical compounds. In addition, these oligosaccharides are employed as anticancer, antioxidant agents and aid in mineral absorption, lipid metabolism, immune regulation etc. This review, therefore, focuses on the occurrence, physico-chemical characteristics, and microbial enzymatic synthesis of FOS and IOS from coprophilous fungi. In addition, the potential health benefits of these oligosaccharides were discussed in detail.

Structural Insight into the Binding of Cyanovirin-N with the Spike Glycoprotein, Mpro and PLpro of SARS-CoV-2: Protein-Protein Interactions, Dynamics Simulations and Free Energy Calculations.

The emergence of COVID-19 continues to pose severe threats to global public health. The pandemic has infected over 171 million people and claimed more than 3.5 million lives to date. We investigated the binding potential of antiviral cyanobacterial proteins including cyanovirin-N, scytovirin and phycocyanin with fundamental proteins involved in attachment and replication of SARS-CoV-2. Cyanovirin-N displayed the highest binding energy scores (-16.8 ± 0.02 kcal/mol, -12.3 ± 0.03 kcal/mol and -13.4 ± 0.02 kcal/mol, respectively) with the spike protein, the main protease (Mpro) and the papainlike protease (PLpro) of SARS-CoV-2. Cyanovirin-N was observed to interact with the crucial residues involved in the attachment of the human ACE2 receptor. Analysis of the binding affinities calculated employing the molecular mechanics-Poisson-Boltzmann surface area (MM-PBSA) approach revealed that all forms of energy, except the polar solvation energy, favourably contributed to the interactions of cyanovirin-N with the viral proteins. With particular emphasis on cyanovirin-N, the current work presents evidence for the potential inhibition of SARS-CoV-2 by cyanobacterial proteins, and offers the opportunity for in vitro and in vivo experiments to deploy the cyanobacterial proteins as valuable therapeutics against COVID-19.

Cyanobacterial metabolites as promising drug leads against the Mpro and PLpro of SARS-CoV-2: an in silico analysis.

A novel severe acute respiratory syndrome coronavirus (SARS-CoV-2) has emerged as the causative agent behind the coronavirus disease 2019 (COVID-19) pandemic. Treatment efforts have been severely impeded due to the lack of specific effective antiviral drugs for the treatment of COVID-associated pathologies. In the present research endeavour the inhibitory prospects of cyanobacterial metabolites were assessed at the active binding pockets of the two vital SARS-CoV-2 proteases namely, main protease (Mpro) and the papain-like protease (PLpro) that proteolytically process viral polyproteins and facilitate viral replication, employing an in silico molecular interaction-based approach. It was evident from our analysis based on the binding energy scores that the metabolites cylindrospermopsin, deoxycylindrospermopsin, carrageenan, cryptophycin 52, eucapsitrione, tjipanazole, tolyporphin and apratoxin A exhibited promising inhibitory potential against the SARS-CoV-2 Mpro. The compounds cryptophycin 1, cryptophycin 52 and deoxycylindrospermopsin were observed to display encouraging binding energy scores with the PLpro of SARS-CoV-2. Subsequent estimation of physicochemical properties and potential toxicity of the metabolites followed by robust molecular dynamics simulations and analysis of MM-PBSA energy scoring function established deoxycylindrospermopsin as the most promising inhibitory candidate against both SARS-CoV-2 proteases. Present research findings bestow ample scopes to further exploit the potential of deoxycylindrospermopsin as a successful inhibitor of SARS-CoV-2 in vitro and in vivo and pave the foundation for the development of novel effective therapeutics against COVID-19.Communicated by Ramaswamy H. Sarma.

Purification and biochemical characterization of an extracellular fructosyltransferase enzyme from Aspergillus niger sp. XOBP48: implication in fructooligosaccharide production.

An extracellular fructosyltransferase (Ftase) enzyme with a molar mass of ≈70 kDa from a newly isolated indigenous coprophilous fungus Aspergillus niger sp. XOBP48 is purified to homogeneity and characterized in this study. The enzyme was purified to 4.66-fold with a total yield of 15.53% and specific activity of 1219.17 U mg-1 of protein after a three-step procedure involving (NH4)2SO4 fractionation, dialysis and anion exchange chromatography. Ftase showed optimum activity at pH 6.0 and temperature 50 °C. Ftase exhibited over 80% residual activity at pH range of 4.0-10.0 and ≈90% residual activity at temperature range of 40-60 °C for 6 h. Metal ion inhibitors Hg2+ and Ag+ significantly inhibited Ftase activity at 1 mmol concentration. Ftase showed K m, v max and k cat values of 79.51 mmol, 45.04 µmol min-1 and 31.5 min-1, respectively, with a catalytic efficiency (k cat/K m) of 396 µmol-1 min-1 for the substrate sucrose. HPLC-RI experiments identified the end products of fructosyltransferase activity as monomeric glucose, 1-kestose (GF2), and 1,1-kestotetraose (GF3). This study evaluates the feasibility of using this purified extracellular Ftase for the enzymatic synthesis of biofunctional fructooligosaccharides.

Monitoring the acclimatization of a Chlorella sp. From freshwater to hypersalinity using photosynthetic parameters of pulse amplitude modulated fluorometry.

Contamination of freshwater raceway ponds impedes the commercial cultivation of microalgae. Acclimatization of freshwater microalgae to hypersaline conditions offers a means to reduce contamination. A freshwater Chlorella species was cultured in a gradient of salinities ranging from 5 to 40‰ and pulse amplitude modulated fluorometry recorded photosynthetic functioning. While the average salinity of seawater is 35‰, optimum acclimatization occurred at 20‰, at which point the growth rate (1.6 µg chl a L-1d-1) was not significantly different from the control (1.8 µg chl a L-1d-1). The maximum relative electron transfer rate was lower (9 to 12 µmol m-2s-1) at 5 to 20‰ as compared to 40‰ (28 µmol m-2s-1) where no algal growth was recorded. ATP and NADPH were thus shunted towards synthesis of molecules that offset cytoplasmic osmotic stress. Culturing this Chlorella strain in raceway ponds under saline conditions may reduce contamination and improve productivity.

Photosystem I fluorescence as a physiological indicator of hydrogen production in Chlamydomonas reinhardtii.

This study investigated the interrelations between hydrogen synthesis and Photosystem I electron transport rate in Chlamydomonas reinhardtii. The fluorescence of both photosystems (PS I and PS II) was monitored using a Dual Pulse Amplitude Modulated (PAM) Fluorometer. Hydrogen synthesis was induced by eliminating sulphur from the growth media (TAP-S). Multiple physiological parameters [rETR, Y (I), Y (II), NPQ, α, Fv/Fm and YI:YII] were recorded using the Dual PAM and correlated to hydrogen produced. There was a 66% increase in Photosystem I rETRmax during hydrogen production. A significant direct correlation existed between PS 1 rETRmax and hydrogen evolution values over the ten-day period (r = 0.895, p < 0.01) indicating that PS I can be considered as a driver of H2 production. Significant correlations between rETRmax of PS I and H2 evolution suggest a novel physiological indicator to monitor H2 production during the three critical phases identified in this study.

Potential biotechnological application of microalgae: a critical review.

Microalgae are diverse microorganisms inhabiting a wide range of habitats with only a small fraction being cultivated for human use. Recently, interest in microalgal research has increased in the quest for alternative renewable fuels due to possible depletion of fossil fuels in the near future. However, costly downstream processing has hampered the commercialization of biofuels derived from microalgae. Several value added products of industrial, pharmaceutical and agricultural relevance could be simultaneously derived from microalgae during bioenergy production. Despite these value-added products having the potential to offset the high cost of downstream processing of renewable fuels, their production has not been explored in-depth. This review presents a critical overview of the current state of biotechnological applications of microalgae for human benefit and highlights possible areas for further research and development.

Perspectives on the probiotic potential of lactic acid bacteria from African traditional fermented foods and beverages.

Diverse African traditional fermented foods and beverages, produced using different types of fermentation, have been used since antiquity because of their numerous nutritional values. Lactic acid bacteria (LAB) isolated from these products have emerged as a welcome source of antimicrobials and therapeutics, and are accepted as probiotics. Probiotics are defined as live microbial food supplements which beneficially affect the host by improving the intestinal microbial balance. Currently, popular probiotics are derived from fermented milk products. However, with the growing number of consumers with lactose intolerance that are affected by dietary cholesterol from milk products, there is a growing global interest in probiotics from other food sources. The focus of this review is to provide an overview of recent developments on the applications of probiotic LAB globally, and to specifically highlight the suitability of African fermented foods and beverages as a viable source of novel probiotics.

The utilization of post-chlorinated municipal domestic wastewater for biomass and lipid production by Chlorella spp. under batch conditions.

South Africa has a rich microalgal biodiversity which has the potential to be used for renewable bio-fuel production in the region. Bioprospecting for oleaginous microalgae in KwaZulu Natal Province, South Africa, resulted in the establishment of a microalgal culture collection system for alternative energy research in the country. A potential hyper-lipid-producing Chlorella spp. strain was isolated, purified, and cultured in supplemented post-chlorinated wastewater for biomass and lipid production at the laboratory scale under batch mode. The microalgal strain was cultivated in different strengths of BG-11 media supplemented with wastewater from a local municipal domestic wastewater treatment plant. The Chlorella spp. was grown using ambient dissolved carbon dioxide in shake flasks under photosynthetically active radiation (±120 µmol m⁻² s⁻¹). Microalgal biomass and lipid productivity were monitored at 24-h intervals in the batch mode. The microalgal biomass was analyzed by direct light microscopy and indirectly by spectrophotometry at 600 nm, and the lipids were extracted and quantified. The growth rate of the Chlorella spp. was enhanced in post-chlorinated wastewater supplemented with 5 mM NaNO3 with maximal biomass productivity. A dramatic increase in lipid yield was achieved with the post-chlorinated wastewater supplemented with 25 mM NaNO3. Low dosages of free chlorine were found to enhance microalgal growth. These findings serve as a basis for further scale-up trials using municipal wastewater as a medium for microalgal biomass and lipid production.