Thermophilic ß-mannanases from bacteria: production, resources, structural features and bioengineering strategies.

ß-mannanases are pivotal enzymes that cleave the mannan backbone to release short chain mannooligosaccharides, which have tremendous biotechnological applications including food/feed, prebiotics and biofuel production. Due to the high temperature conditions in many industrial applications, thermophilic mannanases seem to have great potential to overcome the thermal impediments. Thus, structural analysis of thermostable ß-mannanases is extremely important, as it could open up new avenues for genetic engineering, and protein engineering of these enzymes with enhanced properties and catalytic efficiencies. Under this scope, the present review provides a state-of-the-art discussion on the thermophilic ß-mannanases from bacterial origin, their production, engineering and structural characterization. It covers broad insights into various molecular biology techniques such as gene mutagenesis, heterologous gene expression, and protein engineering, that are employed to improve the catalytic efficiency and thermostability of bacterial mannanases for potential industrial applications. Further, the bottlenecks associated with mannanase production and process optimization are also discussed. Finally, future research related to bioengineering of mannanases with novel protein expression systems for commercial applications are also elaborated.

Enhanced fermentable sugar production in lignocellulosic biorefinery by exploring a novel corn stover and configuring high-solid pretreatment conditions.

This study aimed to produce enhanced fermentable sugars from a novel stover system through the bioprocessing of its soluble sugars and insoluble carbohydrates. The pretreatment conditions were optimized for this high sugar-containing stover (HSS) to control inhibitor formation and obtain enhanced fermentable sugar concentrations. The optimum temperature, acid loading, and reaction time for the pretreatment were 155 °C, 0.5%, and 30 min, respectively, providing up to 97.15% sugar yield and 76.51 g/L total sugars at 10% solid-load. Sugar concentration further increased to 126.9 g/L at 20% solid-load, generating 3.89 g/L acetate, 0.92 g/L 5-hydroxymethyl furfural, 0.82 g/L furfural, and 3.75 g/L total phenolics as inhibitors. To determine the effects of soluble sugars in HSS on fermentable sugar yield and inhibitor formation, sugar-removed HSS was further studied under the optimum conditions. Although prior removal of sugars exhibited a reduction in inhibitor generation, it also decreased total fermentable sugar production to 115.45 g/L.

Host-Specific Diversity of Culturable Bacteria in the Gut Systems of Fungus-Growing Termites and Their Potential Functions towards Lignocellulose Bioconversion.

Fungus-growing termites are eusocial insects that represent one of the most efficient and unique systems for lignocellulose bioconversion, evolved from a sophisticated symbiosis with lignocellulolytic fungi and gut bacterial communities. Despite a plethora of information generated during the last century, some essential information on gut bacterial profiles and their unique contributions to wood digestion in some fungus-growing termites is still inadequate. Hence, using the culture-dependent approach, the present study aims to assess and compare the diversity of lignocellulose-degrading bacterial symbionts within the gut systems of three fungus-growing termites: Ancistrotermes pakistanicus, Odontotermes longignathus, and Macrotermes sp. A total of 32 bacterial species, belonging to 18 genera and 10 different families, were successfully isolated and identified from three fungus-growing termites using Avicel or xylan as the sole source of carbon. Enterobacteriaceae was the most dominant family represented by 68.1% of the total bacteria, followed by Yersiniaceae (10.6%) and Moraxellaceae (9%). Interestingly, five bacterial genera such as Enterobacter, Citrobacter, Acinetobacter, Trabulsiella, and Kluyvera were common among the tested termites, while the other bacteria demonstrated a termite-specific distribution. Further, the lignocellulolytic potential of selected bacterial strains was tested on agricultural waste to evaluate their capability for lignocellulose bioconversion. The highest substrate degradation was achieved with E. chengduensis MA11 which degraded 45.52% of rice straw. All of the potential strains showed endoglucanase, exoglucanase, and xylanase activities depicting a symbiotic role towards the lignocellulose digestion within the termite gut. The above results indicated that fungus-growing termites harbor a diverse array of bacterial symbionts that differ from species to species, which may play an inevitable role to enhance the degradation efficacy in lignocellulose decomposition. The present study further elaborates our knowledge about the termite-bacteria symbiosis for lignocellulose bioconversion which could be helpful to design a future biorefinery.

Nano-Agrochemicals as Substitutes for Pesticides: Prospects and Risks.

This review delves into the mesmerizing technology of nano-agrochemicals, specifically pesticides and herbicides, and their potential to aid in the achievement of UN SDG 17, which aims to reduce hunger and poverty globally. The global market for conventional pesticides and herbicides is expected to reach USD 82.9 billion by 2027, growing 2.7% annually, with North America, Europe, and the Asia-Pacific region being the biggest markets. However, the extensive use of chemical pesticides has proven adverse effects on human health as well as the ecosystem. Therefore, the efficacy, mechanisms, and environmental impacts of conventional pesticides require sustainable alternatives for effective pest management. Undoubtedly, nano-agrochemicals have the potential to completely transform agriculture by increasing crop yields with reduced environmental contamination. The present review discusses the effectiveness and environmental impact of nanopesticides as promising strategies for sustainable agriculture. It provides a concise overview of green nano-agrochemical synthesis and agricultural applications, and the efficacy of nano-agrochemicals against pests including insects and weeds. Nano-agrochemical pesticides are investigated due to their unique size and exceptional performance advantages over conventional ones. Here, we have focused on the environmental risks and current state of nano-agrochemicals, emphasizing the need for further investigations. The review also draws the attention of agriculturists and stakeholders to the current trends of nanomaterial use in agriculture especially for reducing plant diseases and pests. A discussion of the pros and cons of nano-agrochemicals is paramount for their application in sustainable agriculture.

Mining the diversity and functional profile of bacterial symbionts from the larvae of Chironomus circumdatus (bloodworms).

Chironomids are the most abundant aquatic insects in freshwater habitats that can survive in extreme conditions. In this study, as the microbiome provides extended genotype to the host to perform various functions, we explored the microbiota of the Chironomus circumdatus larvae to find out the putative role played by the symbiotic bacteria for the host. The metabarcoding analyses of the larvae revealed that the insect harbors 1771 phylotypes. Out of the various microbial communities found, the majority corresponded to the phyla Proteobacteria (52.59%) and Actinobacteria (20.56%), respectively. The midges also harbored Klebsiella (2.57%), Enterobacter (1.32%), Bacillus (2.29%), and Acinetobacter (2.13%) genera that are involved in detoxification of xenobiotics present in the water. The presence of radiation-resistant genera like Deinococcus, including bacterial species like radiodurans, a highly radiation-resistant bacterium, indicates its potential to support the host's ability to sustain in adverse environments. The functional profiling of the bacteria showed the relative abundance of many enzyme groups, such as transferases (40.62%), oxidoreductases (23.49%), and hydrolases (3.77%). The results indicate that the larvae harbor a considerable variety of bacteria that help the host adapt and survive in the polluted waters. The present study provides thorough insights into the microbiome of the C. circumdatus larvae that can be exploited for the bioremediation of certain pollutants through biomimetic strategies. It also gives us a wake-up call to take a good look at the guts of these disease-carrying insects' inabilities to spread deadly human diseases.

Exploring the region-wise diversity and functions of symbiotic bacteria in the gut system of wood-feeding termite, Coptotermes formosanus, toward the degradation of cellulose, hemicellulose, and organic dyes.

The wood-feeding termite Coptotermes formosanus represents a unique and impressive system for lignocellulose degradation. The highly efficient digestion of lignocellulose is achieved through symbiosis with gut symbionts like bacteria. Despite extensive research during the last three decades, diversity of bacterial symbionts residing in individual gut regions of the termite and their associated functions is still lacking. To this end, cellulose, xylan, and dye-decolorization bacteria residing in foregut, midgut, and hindgut regions of C. formosanus were enlisted by using enrichment and culture-dependent molecular methods. A total of 87 bacterial strains were successfully isolated from different gut regions of C. formosanus which belonged to 27 different species of 10 genera, majorly affiliated with Proteobacteria (80%) and Firmicutes (18.3%). Among the gut regions, 37.9% of the total bacterial isolates were observed in the hindgut that demonstrated predominance of cellulolytic bacteria (47.6%). The majority of the xylanolytic and dye-decolorization bacteria (50%) were obtained from the foregut and midgut, respectively. Actinobacteria represented by Dietza sp. was observed in the hindgut only. Based on species richness, the highest diversity was observed in midgut and hindgut regions each of which harbored seven unique bacterial species. The members of Enterobacter, Klebsiella, and Pseudomonas were common among the gut regions. The lignocellulolytic activities of the selected potential bacteria signpost their assistance to the host for lignocellulose digestion. The overall results indicate that C. formosanus harbors diverse communities of lignocellulolytic bacteria in different regions of the gut system. These observations will significantly advance our understanding of the termite-bacteria symbiosis and their microbial ecology uniquely existed in different gut regions of C. formosanus, which may further shed a light on its potential values at termite-modeled biotechnology.

Detection and molecular characterization of picobirnaviruses in the wild birds: Identification of a novel picobirnavirus possessing yeast mitochondrial genetic code.

Picobirnaviruses (PBVs) are bi-segmented dsRNA viruses that have been detected in various animal species including vertebrates and invertebrates. In this study, 17 complete or incomplete PBV segment-2 and one unsegmented PBV-like virus sequence were identified in fecal samples from different bird species using viral metagenomic approach. The bird PBV and PBV-like virus retained the conservative motifs that are conserved in dsRNA2 of common PBVs. The RdRp of these 17 PBVs shared the highest Amino acid (aa) identity of 45.90%∼94.19% with previous animal and human PBVs, while the RdRp of the unsegment PBV-like virus shared the highest aa sequence identity of 31.93% with one chicken PBV (GenBank No. MW837829). The unsegmented PBV-like virus unexpectedly used the yeast mitochondrial genetic code (transl_table=3) for all ORFs translation. In addition, the prokaryotic RBS sequence was not only detected upstream to ORF2 at position 360AGGAGG365 of this unsegmented PBV-like virus, but also found upstream to ORF of bird PBV dsRNA2. The presence of the prokaryotic ribosomal binding site in the bird PBV genomes, and the finding of one novel unsegmented PBV-like virus using the yeast mitochondrial genetic code for translation supported recent speculations that PBVs may actually infect prokaryotic or fungal host cells. This study enhanced our understanding of PBVs and provided data support for exploring the real host of PBVs.

Valorization Potential of a Novel Bacterial Strain, Bacillus altitudinis RSP75, towards Lignocellulose Bioconversion: An Assessment of Symbiotic Bacteria from the Stored Grain Pest, Tribolium castaneum.

Bioconversion of lignocellulose into renewable energy and commodity products faces a major obstacle of inefficient saccharification due to its recalcitrant structure. In nature, lignocellulose is efficiently degraded by some insects, including termites and beetles, potentially due to the contribution from symbiotic gut bacteria. To this end, the presented investigation reports the isolation and characterization of cellulolytic bacteria from the gut system of red flour beetle, Tribolium castaneum. Out of the 15 isolated bacteria, strain RSP75 showed the highest cellulolytic activities by forming a clearance zone of 28 mm in diameter with a hydrolytic capacity of ~4.7. The MALDI-TOF biotyping and 16S rRNA gene sequencing revealed that the strain RSP75 belongs to Bacillus altitudinis. Among the tested enzymes, B. altitudinis RSP75 showed maximum activity of 63.2 IU/mL extract for xylanase followed by ß-glucosidase (47.1 ± 3 IU/mL extract) which were manifold higher than previously reported activities. The highest substrate degradation was achieved with wheat husk and corn cob powder which accounted for 69.2% and 54.5%, respectively. The scanning electron microscopy showed adhesion of the bacterial cells with the substrate which was further substantiated by FTIR analysis that depicted the absence of the characteristic cellulose bands at wave numbers 1247, 1375, and 1735 cm-1 due to hydrolysis by the bacterium. Furthermore, B. altitudinis RSP75 showed co-culturing competence with Saccharomyces cerevisiae for bioethanol production from lignocellulose as revealed by GC-MS analysis. The overall observations signify the gut of T. castaneum as a unique and impressive reservoir to prospect for lignocellulose-degrading bacteria that can have many biotechnological applications, including biofuels and biorefinery.

Recent advances in the bioprospection and applications of chitinolytic bacteria for valorization of waste chitin.

One of the most abundant natural polymers on earth, chitin is a fibrous and structural polysaccharide, composed of N-acetyl-D-glucosamine. The biopolymer is the major structural constituent of fungi, arthropods, mollusks, nematodes, and some algae. The biodegradation of chitin is largely manifested by chitinolytic enzyme secreting organisms including bacteria, insects, and plants. Among them, bacterial chitinases represent the most promising, inexpensive, and sustainable source of proteins that can be employed for industrial-scale applications. To this end, the presented review comes at a timely moment to highlight the major sources of chitinolytic bacteria. It also discusses the potential pros and cons of prospecting bacterial chitinases that can be easily manipulated through genetic engineering. Additionally, we have elaborated the recent applications of the chitin thereby branding chitinases as potential candidates for biorefinery and biomedical research for eco-friendly and sustainable management of chitin waste in the environment.

Next Generation High Throughput Sequencing to Assess Microbial Communities: An Application Based on Water Quality.

Traditional techniques to identify different contaminants (biological or chemical) in the waters are slow, laborious, and can require specialized expertise. Hence, the rapid determination of water quality using more sensitive and reliable metagenomic based approaches attains special importance. Metagenomics deals with the study of genetic material that is recovered from microbial communities present in environmental samples. In traditional techniques cultivation-based methodologies were used to describe the diversity of microorganisms in environmental samples. It has failed to function as a robust marker because of limited taxonomic and phylogenetic implications. In this backdrop, high-throughput DNA sequencing approaches have proven very powerful in microbial source tracking because of investigating the full variety of genome-based analysis such as microbial genetic diversity and population structure played by them. Next generation sequencing technologies can reveal a greater proportion of microbial communities that have not been reported earlier by traditional techniques. The present review highlights the shift from traditional techniques for the basic study of community composition to next-generation sequencing (NGS) platforms and their potential applications to the biomonitoring of water quality in relation to human health.

Bioefficacy of Lemongrass and Tea Tree Essential Oils Against House Fly, Musca domestica.

ABSTRACT: The management of house fly, Musca domestica has become immensely important to prevent epidemics of many detrimental diseases. In view of this objective, the present study demonstrates the efficacy of lemongrass (LG) and tea tree essential oils (TTEOs) against M. domestica. The TTEO proved to be more lethal against larvae and adults of M. domestica depicting an LC50 at 14.88 mg/ dm3 which was 17.19 mg/ dm3 for LGEO. In contrast, pupicidal effect of LGEO was much higher (LD50, 14.49 µl/0.25L) as compared to TTEO. The LGEO drastically reduced the total body sugar, glycogen and protein contents by 3.29, 2.95 and 7.56 fold, respectively, contrasting with high influence of TTEO on lipid content of the late 3rd instar larvae. A considerable reduction in gut enzymes secretion was observed due to treatment of EOs thereby altering gut physiology of the insect. Moreover, significant inhibition of acetylcholine esterase (AchE) was also observed with LGEO at LC50 concentration (5.33 mg/ml) inhibiting insect neurotransmission. The gas chromatography-mass spectrometric (GC-MS) analysis of the LGEO showed 12 major compounds dominated by Citral whereas TTEO contained only 5 major compounds. Further analysis by field emission scanning electron microscopy (FESEM) revealed distortion and shrinkage of larval bodies caused by the treatment of EOs. These overall observations brand LG and TT-EOs as potential organic-insecticides against M. domestica. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s40011-020-01220-z) contains supplementary material, which is available to authorized users.

Enrichment and identification of cellulolytic bacteria from the gastrointestinal tract of Giant African snail, Achatina fulica.

The cellulolytic bacterial community structure in gastrointestinal (GI) tract of Achatina fulica was studied using culture-independent and -dependent methods by enrichment in carboxymethyl cellulose (CMC). Culture-dependent method indicated that GI tract of snail was dominated by Enterobacteriaceae members. When tested for cellulase activities, all isolates obtained by culture-dependent method showed both or either of CMCase or avicelase activity. Isolate identified as Citrobacter freundii showed highest CMCase and medium avicelase activity. Sequencing of clones from the 16S rRNA gene clone library identified ten operational taxonomic units (OTUs), which were affiliated to Enterobacteriaceae of phylum Gammaproteobacteria. Of these ten OTUs, eight OTUs closely matched with Enterobacter and Klebsiella genera. The most abundant OTU allied to Klebsiella oxytoca accounted for 70 % of the total sequences. The members of Klebsiella and Enterobacter were observed by both methods indicating their dominance among the cellulolytic bacterial community in the GI tract of the snail.

Enhanced antimicrobial activity of silver nanoparticles synthesized by Cryphonectria sp. evaluated singly and in combination with antibiotics.

The newly identified pathogenic species of the genus Cryphonectria was exploited for the synthesis of silver nanoparticles (AgNPs). The extracellular synthesis of nanoparticles was fast and eco-friendly. These nanoparticles were analyzed and determined by ultraviolet-visible spectrophotometry, which detected AgNPs in the solution, and by NanoSight LM20, which determined their average size and concentration. The synthesized AgNPs showed a little monodispersity in the range of 30-70 nm with a concentration of 6.82 × 10(8) particles per milliliter of solution. The AgNPs demonstrated antibacterial activity against Staphylococcus aureus, Escherichia coli, Salmonella typhi, and Candida albicans, with comparatively higher activity against both S. aureus and E. coli than against S. typhi and C. albicans, which showed the lowest activity. The present study demonstrates the possible use of biologically synthesized AgNPs in the field of medicine. Thus, Cryphonectria sp. could be used for simple, nonhazardous, and efficient synthesis of AgNPs. FROM THE CLINICAL EDITOR: In this study Cryphonectria sp. was exploited for simple, nonhazardous, and efficient biosynthesis of silver nanopartricles, which had antibacterial activity against a variety of human pathogenic bacteria.