Beauveria Bassiana Amylase-Polygalacturonase Production Using Lignocellulosic Biomass and Application in Juice Processing.

Background: The search for sources of industrial biocatalysts, which are non-pathogenic and can utilise cheap nutrient sources, has been a continuous endeavour in the ~ 7 billion USD enzyme industry. Beauveria bassiana, an endophytic fungal entomopathogen, is non-pathogenic and possesses the potential to secrete various bioproducts while utilising readily available lignocellulosic biomass. Objective: This study investigated the optimised production of two glycosyl hydrolases, amylase and polygalacturonase, by B. bassiana while utilising readily available agricultural residues. Subsequently, the industrial potential of the enzymes in the clarification of fruit juice was evaluated. Materials and Methods: Initially, seven agro residues were screened for the concomitant production of amylase and polygalacturonase by B. bassiana SAN01. Subsequently, statistical optimisation tools, Plackett Burman Design (PBD) and Central Composite Design (CCD), were employed for the optimisation of enzyme production. The enzyme mixture was partially purified and applied in the clarification of pineapple juice. Result: The production of B. bassiana SAN01 amylase and polygalacturonase was found to be maximal while utilising wheat bran. Subsequent to PBD and CCD optimisation, the optimal conditions for enzyme production were identified to be at 30 °C, pH 6.0 and wheat bran concentration of ~40 g.L-1. Under these optimised conditions, heightened production levels of 34.82 and 51.05 U.mL-1 were recorded for amylase and polygalacturonase, respectively, which were 179% and 187% of the initial unoptimised levels. In addition, the most effective clarification of the juice (~90%) was observed at 35 °C after an incubation time of 120 min with no significant effect on the pH and total dissolved solids. Conclusion: B. bassiana, a well-known biocontrol agent, was shown to produce amylase and polygalacturonase using readily available agricultural residues for the first time. These enzyme production levels are the highest for these enzymes from any known endophytic fungal entomopathogen. This study further demonstrates the potential applicability of B. bassiana in other industrial processes besides its widespread use as a biopesticide.

Microbial ß-glucosidases: Recent advances and applications.

The global ß-glucosidase market is currently estimated at ∼400 million USD, and it is expected to double in the next six years; a trend that is mainly ascribed to the demand for the enzyme for biofuel processing. Microbial ß-glucosidase, particularly, has thus garnered significant attention due to its ease of production, catalytic efficiency, and versatility, which have all facilitated its biotechnological potential across different industries. Hence, there are continued efforts to screen, produce, purify, characterize and evaluate the industrial applicability of ß-glucosidase from actinomycetes, bacteria, fungi, and yeasts. With this rising demand for ß-glucosidase, various cost-effective and efficient approaches are being explored to discover, redesign, and enhance their production and functional properties. Thus, this present review provides an up-to-date overview of advancements in the utilization of microbial ß-glucosidases as "Emerging Green Tools" in 21st-century industries. In this regard, focus was placed on the use of recombinant technology, protein engineering, and immobilization techniques targeted at improving the industrial applicability of the enzyme. Furthermore, insights were given into the recent progress made in conventional ß-glucosidase production, their industrial applications, as well as the current commercial status-with a focus on the patents.

Biochemical and in silico structural properties of a thermo-acid stable ß-glucosidase from Beauveria bassiana.

ß-glucosidase hydrolyses the glycosidic bonds in cellobiose and cello-oligosaccharides, a critical step in the saccharification for biofuel production. Hence, the aim of this study was to gain insights into the biochemical and structural properties of a ß-glucosidase from Beauveria bassiana, an entomopathogenic fungus. The ß-glucosidase was purified to homogeneity using salt precipitation, ultrafiltration, and chromatographic techniques, attaining a specific activity of 496 U/mg. The molecular mass of the enzyme was then estimated via SDS-PAGE to be 116 kDa, while its activity pattern was confirmed by zymography using 4-methylumbelliferyl-ß-d-glucopyranoside. Furthermore, the pH optima and temperature of the enzyme were found to be pH 5.0 and 60 °C respectively; its activity was significantly enhanced by Mg2+ and Na+ and was found to be relatively moderate in the presence of ethanol and dichloromethane. Molecular docking of the modelled B. bassiana ß-glucosidase structure with the substrates, viz., 4-nitrophenyl ß-d-glucopyranoside and cellobiose, revealed the binding affinity energies of -7.2 and -6.2 (kcal mol-1), respectively. Furthermore, the computational study predicted Lys-657, Asp-658, and Arg-1000 as the core amino acid residues in the catalytic site of the enzyme. This is the first investigation into a purified ß-glucosidase from B. bassiana, providing valuable insights into the functional properties of carbohydrases from entomopathogenic fungal endophytes.

Biochemical engineering for elemental sulfur from flue gases through multi-enzymatic based approaches - A review.

Flue gases are the gases which are produced from industries related to chemical manufacturing, petrol refineries, power plants and ore processing plants. Along with other pollutants, sulfur present in the flue gas is detrimental to the environment. Therefore, environmentalists are concerned about its removal and recovery of resources from flue gases due to its activation ability in the atmosphere to transform into toxic substances. This review is aimed at a critical assessment of the techniques developed for resource recovery from flue gases. The manuscript discusses various bioreactors used in resource recovery such as hollow fibre membrane reactor, rotating biological contractor, sequential batch reactor, fluidized bed reactor, entrapped cell bioreactor and hybrid reactors. In conclusion, this manuscript provides a comprehensive analysis of the potential of thermotolerant and thermophilic microbes in sulfur removal. Additionally, it evaluates the efficacy of a multi-enzyme engineered bioreactor in this process. Furthermore, the study introduces a groundbreaking sustainable model for elemental sulfur recovery, offering promising prospects for environmentally-friendly and economically viable sulfur removal techniques in various industrial applications.

Writing a Scientific Review Article: Comprehensive Insights for Beginners.

Review articles present comprehensive overview of relevant literature on specific themes and synthesise the studies related to these themes, with the aim of strengthening the foundation of knowledge and facilitating theory development. The significance of review articles in science is immeasurable as both students and researchers rely on these articles as the starting point for their research. Interestingly, many postgraduate students are expected to write review articles for journal publications as a way of demonstrating their ability to contribute to new knowledge in their respective fields. However, there is no comprehensive instructional framework to guide them on how to analyse and synthesise the literature in their niches into publishable review articles. The dearth of ample guidance or explicit training results in students having to learn all by themselves, usually by trial and error, which often leads to high rejection rates from publishing houses. Therefore, this article seeks to identify these challenges from a beginner's perspective and strives to plug the identified gaps and discrepancies. Thus, the purpose of this paper is to serve as a systematic guide for emerging scientists and to summarise the most important information on how to write and structure a publishable review article.

The potential of fungi in the bioremediation of pharmaceutically active compounds: a comprehensive review.

The ability of fungal species to produce a wide range of enzymes and metabolites, which act synergistically, makes them valuable tools in bioremediation, especially in the removal of pharmaceutically active compounds (PhACs) from contaminated environments. PhACs are compounds that have been specifically designed to treat or alter animal physiological conditions and they include antibiotics, analgesics, hormones, and steroids. Their detrimental effects on all life forms have become a source of public outcry due their persistent nature and their uncontrolled discharge into various wastewater effluents, hospital effluents, and surface waters. Studies have however shown that fungi have the necessary metabolic machinery to degrade PhACs in complex environments, such as soil and water, in addition they can be utilized in bioreactor systems to remove PhACs. In this regard, this review highlights fungal species with immense potential in the biodegradation of PhACs, their enzymatic arsenal as well as the probable mechanism of biodegradation. The challenges encumbering the real-time application of this promising bioremediative approach are also highlighted, as well as the areas of improvement and future perspective. In all, this paper points researchers to the fact that fungal bioremediation is a promising strategy for addressing the growing issue of pharmaceutical contamination in the environment and can help to mitigate the negative impacts on ecosystems and human health.

Current research trends on cosmetic microplastic pollution and its impacts on the ecosystem: A review.

Since the advent of microplastics, it has become a vital component, directly or indirectly, in our daily lives. With advancements in their use, microplastics have become an integral part of personal care, cosmetics, and cleaning products (PCCPs) and emerged as a domestic source of environmental pollution. Over the years, researchers have ascertained the harmful effects of microplastics on the environment. In this context, the assessment and monitoring of microplastics in PCCPs require considerable attention. In addition, it raises concern regarding the need to develop innovative, sustainable, and environmentally safe technologies to combat microplastic pollution. Therefore, this review is an endeavor to uncover the fate, route and degradation mechanism of cosmetic microplastics. In addition, the major technological advancement in cosmetic microplastic removal and the steps directed toward mitigating cosmetic microplastic pollution are also discussed.

Beauveria bassiana Xylanase: Characterization and Wastepaper Deinking Potential of a Novel Glycosyl Hydrolase from an Endophytic Fungal Entomopathogen.

Beauveria bassiana is an entomopathogenic fungus widely used as a biopesticide for insect control; it has also been shown to exist as an endophyte, promoting plant growth in many instances. This study highlights an alternative potential of the fungus; in the production of an industrially important biocatalyst, xylanase. In this regard, Beauveria bassiana SAN01 xylanase was purified to homogeneity and subsequently characterized. The purified xylanase was found to have a specific activity of 324.2 U·mg-1 and an estimated molecular mass of ~37 kDa. In addition, it demonstrated optimal activity at pH 6.0 and 45 °C while obeying Michaelis-Menton kinetics towards beechwood xylan with apparent Km, Vmax and kcat of 1.98 mg·mL-1, 6.65 µM·min-1 and 0.62 s-1 respectively. The enzyme activity was strongly inhibited by Ag2+ and Fe3+ while it was significantly enhanced by Co2+ and Mg2+. Furthermore, the xylanase was shown to effectively deink wastepaper at an optimal rate of 106.72% through its enzymatic disassociation of the fiber-ink bonds as demonstrated by scanning electron microscopy and infrared spectroscopy. This is the first study to demonstrate the biotechnological application of a homogeneously purified glycosyl hydrolase from B. bassiana.

Characterisation, pathogenicity and hydrolytic enzyme profiling of selected Fusarium species and their inhibition by novel coumarins.

Three Fusarium species isolated locally were characterised by the amplification of their rDNA ITS region, host specificity, and hydrolytic enzyme production. The strains were identified as Fusarium pseudoanthophilum, which is being reported for the first time in South Africa, as well as F. foetens and F. fujikuroi. All the three strains were capable of infecting vegetables such as tomatoes, bell and cayenne peppers, belonging to the Solanaceae family. The Fusarium strains also showed significant production of cell wall degrading enzymes in vitro, such as amylase, cellulase, xylanase, and polygalacturonase, thus highlighting the possibilities of these enzymes as pathogenic factors. Subsequently, the strains were discovered to be susceptible to three halogenated coumarins. The most effective of the tested coumarins, 6-bromo3-2,2-dibromoacetyl-2H-chromen-2-one, showed MIC values of 0.125, 0.0625 and 0.125 mg/ml against F. foetens, F. pseudoanthophilum and F. fujikuroi, respectively. The antifungal potentials of the halogenated coumarins were confirmed in silico through PASS analysis, toxicity prediction and docking studies. Findings from this study demonstrate the use of these coumarins as potential control agents against the Fusarium species and other pathogenic fungi in general.

Enhanced xylanase and endoglucanase production from Beauveria bassiana SAN01, an entomopathogenic fungal endophyte.

This study was undertaken to explore alternative applications of the widely known entomopathogenic/endophytic fungus, Beauveria bassiana, besides its sole use as a biocontrol agent. B. bassiana SAN01, was investigated for the production of two glycoside hydrolases, xylanase and endoglucanase under submerged conditions. Among the different biomass tested, wheat bran provided the best results for both xylanase and endoglucanase, and their production levels were further enhanced using response surface methodology. Under optimised conditions, heightened yields of 1061 U/ml and 23.03 U/ml were observed for xylanase and endoglucanase, respectively, which were 3.44 and 1.35 folds higher than their initial yields. These are the highest ever production levels reported for xylanase and endoglucanase from any B. bassiana strain or any known entomopathogenic fungi. Furthermore, the efficacy of xylanase/endoglucanase cocktail in the saccharification of sugarcane bagasse was evaluated. The highest amount of reducing sugar released from the pretreated biomass by the action of the crude Beauveria enzyme cocktail was recorded at 30°C after 8 h incubation. The significant activities of the hydrolytic enzymes recorded with B. bassiana in this study thus present promising avenues for the use of the entomopathogen as a new source of industrial enzymes and by extension, other biotechnological applications.

Environmental Impacts of Microplastics and Nanoplastics: A Current Overview.

The increasing distribution of miniaturized plastic particles, viz. microplastics (100 nm-5 mm) and nanoplastics (less than 100 nm), across the various ecosystems is currently a subject of major environmental concern. Exacerbating these concerns is the fact that microplastics and nanoplastics (MNPs) display different properties from their corresponding bulk materials; thus, not much is understood about their full biological and ecological implications. Currently, there is evidence to prove that these miniaturized plastic particles release toxic plastic additives and can adsorb various chemicals, thereby serving as sinks for various poisonous compounds, enhancing their bioavailability, toxicity, and transportation. Furthermore, there is a potential danger for the trophic transfer of MNPs to humans and other higher animals, after being ingested by lower organisms. Thus, this paper critically analyzes our current knowledge with regard to the environmental impacts of MNPs. In this regard, the properties, sources, and damaging effects of MNPs on different habitats, particularly on the biotic components, were elucidated. Similarly, the consequent detrimental effects of these particles on humans as well as the current and future efforts at mitigating these detrimental effects were discussed. Finally, the self-cleaning efforts of the planet via a range of saprophytic organisms on these synthetic particles were also highlighted.

Plastic biodegradation: Frontline microbes and their enzymes.

Plastic polymers with different properties have been developed in the last 150 years to replace materials such as wood, glass and metals across various applications. Nevertheless, the distinct properties which make plastic desirable for our daily use also threaten our planet's sustainability. Plastics are resilient, non-reactive and most importantly, non-biodegradable. Hence, there has been an exponential increase in plastic waste generation, which has since been recognised as a global environmental threat. Plastic wastes have adversely affected life on earth, primarily through their undesirable accumulation in landfills, leaching into the soil, increased greenhouse gas emission, etc. Even more damaging is their impact on the aquatic ecosystems as they cause entanglement, ingestion and intestinal blockage in aquatic animals. Furthermore, plastics, especially in the microplastic form, have also been found to interfere with chemical interaction between marine organisms, to cause intrinsic toxicity by leaching, and by absorbing persistent organic contaminants as well as pathogens. The current methods for eliminating these wastes (incineration, landfilling, and recycling) come at massive costs, are unsustainable, and put more burden on our environment. Thus, recent focus has been placed more on the potential of biological systems to degrade synthetic plastics. In this regard, some insects, bacteria and fungi have been shown to ingest these polymers and convert them into environmentally friendly carbon compounds. Hence, in the light of recent literature, this review emphasises the multifaceted roles played by microorganisms in this process. The current understanding of the roles played by actinomycetes, algae, bacteria, fungi and their enzymes in enhancing the degradation of synthetic plastics are reviewed, with special focus on their modes of action and probable enzymatic mechanisms. Besides, key areas for further exploration, such as the manipulation of microorganisms through molecular cloning, modification of enzymatic characteristics and metabolic pathway design, are also highlighted.

Biotechnological potential of Beauveria bassiana as a source of novel biocatalysts and metabolites.

Beauveria bassiana though widely perceived as an entomopathogenic fungus has also been found in nature to be endophytic. As entomopathogens, the life cycle of different B. bassiana strains are organized and adapted as pathogens to their invertebrate hosts while as endophytes they maintain a symbiotic relationship with their plant hosts. To fulfill these aforementioned ecological roles, this fungus secretes an array of enzymes as well as secondary metabolites, which all have significant biological roles. Basically, chitinases, lipases and proteases are considered to be the most important of all the enzymes produced by B. bassiana. However, studies have also shown their ability to produce other vital enzymes which include amylase, asparaginase, cellulase, galactosidase etc. Previous reports on this filamentous fungus have laid more emphasis on its entomopathogenicity, its endophytism and its highly acclaimed application in the biological control of pests. This review, however, is the first to fully assess the enzyme-secreting potential of this entomopathogenic fungus and its use as a novel source of several industrial biocatalysts and other important biochemicals. This article highlights the inherent properties of the fungus to degrade various biopolymers as well as its relative safety for human use. Some of the important factors have raised the possibilities of exploitation for industrial production and as safe hosts for gene expression.

Recent advances in microbial glutaminase production and applications-a concise review.

This article focuses on significant advances in the production and applications of microbial glutaminases and provides insight into the structures of different glutaminases. Glutaminases catalyze the deamidation of glutamine to glutamic acid, and this unique ability forms the basis of their applications in various industries such as pharmaceutical and food organizations. Microbial glutaminases from bacteria, actinomycetes, yeast, and fungi are of greater significance than animal glutaminases because of their stability, affordability, and ease of production. Owing to these notable benefits, they are considered to possess considerable potential in anticancer and antiviral therapy, flavor enhancers in oriental foods, biosensors and in the production of a nutraceutical theanine. This review also aims to fully explore the potential of microbial glutaminases and to set the pace for future prospects.

Protective effects of Parinari curatellifolia flavonoids against acetaminophen-induced hepatic necrosis in rats.

In the present study, we investigated the hepatoprotective potential of Parinari curatellifolia Planch (Chrysobalanaceae) in experimental rats in order to ascertain the validity of folkloric claims of its effectiveness in the treatment of hepatic-related disorders. Flavonoid extract of P. curatellifolia seed, PCF (10-, 20- or 30 mg/kg body weight) or silymarin (25 mg/kg), dissolved in corn oil, was administered by gavage to experimental animals once daily for 14 consecutive days before liver damage was chemically induced through the administration of acetaminophen (2 g/kg p.o.) on the 14th day. Hepatoprotection was assessed by analyzing liver homogenate and serum for markers of hepatotoxicity - alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transferase (GGT) and lactate dehydrogenase (LDH) activities as well as prothrombin time (PT). Evaluation of biochemical indices of oxidative stress - level of lipid peroxides (LPO), activities of superoxide dismutase (SOD) and catalase, along with histological assessment of hepatic tissue sections were also carried out. Results revealed that all doses of PCF significantly (P < 0.001) and dose dependently prevented acetaminophen-induced increase in serum activities of hepatic enzymes (ALT, AST, GGT, LDH) and PT. Furthermore, PCF (10- and 20 mg/kg) significantly (P < 0.001) reduced lipid peroxidation in liver tissue and restored the activities of the antioxidant enzymes SOD and catalase toward normal levels. Histopathology of the liver tissue showed that PCF mitigated the toxicant-induced hepatocellular necrosis, reduced inflammatory cell infiltration and enhanced hepatocyte regeneration. The results indicated that P. curatellifolia flavonoids demonstrated remarkable hepatoprotective activity in acute liver injury caused by acetaminophen.