Antimicrobial Activity of the Secondary Metabolites Isolated from a South African Red Seaweed, Laurencia corymbosa.

South Africa's highly diverse marine biota includes several endemic marine red algae of the Laurencia genus. Cryptic species and morphological variability make the taxonomy of Laurencia plant challenging, and a record of the secondary metabolites isolated from South African Laurencia spp. can be used to assess their chemotaxonomic significance. In addition, the rapid development of resistance against antibiotics, coupled with the inherent ability of seaweeds to resist pathogenic infection, supported this first phycochemical investigation of Laurencia corymbosa J. Agardh. A new tricyclic keto-cuparane (7) and two new cuparanes (4, 5) were obtained alongside known acetogenins, halo-chamigranes, and additional cuparanes. These compounds were screened against Acinetobacter baumannii, Enterococcus faecalis, Escherichia coli, Staphylococcus aureus, and Candida albicans, with 4 exhibiting excellent activity against the Gram-negative A. baumanii (minimum inhibitory concentration (MIC) 1 µg/mL) strain.

Peroxidase-producing actinobacteria from Algerian environments and insights from the genome sequence of peroxidase-producing Streptomyces sp. S19.

Unique environments often serve as a source of novel microorganisms with novel chemistries. In this study, telluric samples collected from different regions of Algeria were processed for the isolation of novel peroxidase-producing actinobacterial strains. An agar-based screening identified 45 isolates with the ability to produce peroxidase. The 16S rRNA gene sequencing showed that most of the strains belong to the genus Streptomyces. Optimization of cultivation conditions was performed for the top five peroxidase-producing strains. Apart from strain 36 (optimal growth temperature of 30 °C) and strain 45 (optimal medium pH of 6.0), the strains exhibited optimal peroxidase production when cultivated for 5 days at 37 °C and in a medium at pH 7.0. Extracellular peroxidase production was induced by ferulic acid in three of the five strains, while the presence of canola lignocellulosic waste (CLW) induced peroxidase production in all strains. Strain 19 (S19) was selected for further optimization and the extracellular peroxidase purified using acid and acetone precipitation, followed by size exclusion chromatography. The purified fraction showed a single band on the polyacrylamide gel with an estimated molecular weight of 21.45 kDa. Genome analysis confirmed the assignment of S19 to the genus Streptomyces, the presence of genes encoding for peroxidases, and the presence of genes encoding for carbohydrate-active enzymes. The presence of biosynthetic gene clusters potentially encoding for biosurfactants further highlighted the great biotechnological potential of the strain.

Flavonoid and Phenolic Acid Profiles of Dehulled and Whole Vigna subterranea (L.) Verdc Seeds Commonly Consumed in South Africa.

Bambara groundnut (BGN) is an underexploited crop with a rich nutrient content and is used in traditional medicine, but limited information is available on the quantitative characterization of its flavonoids and phenolic acids. We investigated the phenolic profile of whole seeds and cotyledons of five BGN varieties consumed in South Africa using UPLC-qTOF-MS and GC-MS. Twenty-six phenolic compounds were detected/quantified in whole seeds and twenty-four in cotyledon, with six unidentified compounds. Flavonoids include flavan-3-ol (catechin, catechin hexoside-A, catechin hexoside-B), flavonol (quercetin, quercetin-3-O-glucoside, rutin, myricetin, kaempherol), hydroxybenzoic acid (4-Hydroxybenzoic, 2,6 Dimethoxybenzoic, protocatechuic, vanillic, syringic, syringaldehyde, gallic acids), hydroxycinnamic acid (trans-cinnamic, p-coumaric, caffeic, ferulic acids) and lignan (medioresinol). The predominant flavonoids were catechin/derivatives, with the highest content (78.56 mg/g) found in brown BGN. Trans-cinnamic and ferulic acids were dominant phenolic acid. Cotyledons of brown and brown-eyed BGN (317.71 and 378.59 µg/g) had the highest trans-cinnamic acid content, while red seeds had the highest ferulic acid (314.76 µg/g) content. Colored BGN had a significantly (p < 0.05) higher content of these components. Whole BGN contained significantly (p < 0.05) higher amount of flavonoids and phenolic acids, except for the trans-cinnamic acid. The rich flavonoid and phenolic acid content of BGN seeds highlights the fact that it is a good source of dietary phenolics with potential health-promoting properties.

Expansin Engineering Database: A navigation and classification tool for expansins and homologues.

Expansins have the remarkable ability to loosen plant cell walls and cellulose material without showing catalytic activity and therefore have potential applications in biomass degradation. To support the study of sequence-structure-function relationships and the search for novel expansins, the Expansin Engineering Database (ExED, https://exed.biocatnet.de) collected sequence and structure data on expansins from Bacteria, Fungi, and Viridiplantae, and expansin-like homologues such as carbohydrate binding modules, glycoside hydrolases, loosenins, swollenins, cerato-platanins, and EXPNs. Based on global sequence alignment and protein sequence network analysis, the sequences are highly diverse. However, many similarities were found between the expansin domains. Newly created profile hidden Markov models of the two expansin domains enable standard numbering schemes, comprehensive conservation analyses, and genome annotation. Conserved key amino acids in the expansin domains were identified, a refined classification of expansins and carbohydrate binding modules was proposed, and new sequence motifs facilitate the search of novel candidate genes and the engineering of expansins.

Application of bacterial tyrosinases in organic synthesis.

Bacterial tyrosinases, as in the case of other bacterial oxidative enzymes, have been found to possess biochemical characteristics that typically make them more suited to applications requiring special operational conditions such as alkaline pH, high or low temperature, the presence of organic solvents, and the presence of inhibitors. Even though a great deal is known about fungal tyrosinases, bacterial tyrosinases still vastly remain underexplored for their potential application in organic synthesis. A literature survey in particular highlights the gaps in our knowledge pertaining to their biochemical properties. Bacterial tyrosinases have not only shown promise in the synthesis of medically important compounds such as L-3,4-dihydroxyphenylalanine (L-DOPA) and melanin but have also seen application in cross-linking reactions of proteins and the polymerization of environmental pollutants. Their ability to catalyse o-hydroxylation reactions have shown some degree of promise in the biocatalytic conversion of resveratrol to piceatannol, tyrosol to hydroxytyrosol, and many more. In this review, we will explore the world of bacterial tyrosinases, their current applications, and future perspectives for the application of these enzymes in organic synthesis.

Multicopper oxidases: modular structure, sequence space, and evolutionary relationships.

Multicopper oxidases (MCOs) use copper ions as cofactors to oxidize a variety of substrates while reducing oxygen to water. MCOs have been identified in various taxa, with notable occurrences in fungi. The role of these fungal MCOs in lignin degradation sparked an interest due to their potential for application in biofuel production and various other industries. MCOs consist of different protein domains, which led to their classification into two-, three-, and six-domain MCOs. The previously established Laccase and Multicopper Oxidase Engineering Database (https://lcced.biocatnet.de) was updated and now includes 51 058 sequences and 229 structures of MCOs. Sequences and structures of all MCOs were systematically compared. All MCOs consist of cupredoxin-like domains. Two-domain MCOs are formed by the N- and C-terminal domain (domain N and C), while three-domain MCOs have an additional domain (M) in between, homologous to domain C. The six-domain MCOs consist of alternating domains N and C, each three times. Two standard numbering schemes were developed for the copper-binding domains N and C, which facilitated the identification of conserved positions and a comparison to previously reported results from mutagenesis studies. Two sequence motifs for the copper binding sites were identified per domain. Their modularity, depending on the placement of the T1-copper binding site, was demonstrated. Protein sequence networks showed relationships between two- and three-domain MCOs, allowing for family-specific annotation and inference of evolutionary relationships.

Leptospermum petersonii as a Potential Natural Food Preservative.

Leptospermum petersonii (family Myrtaceae) is often cultivated for ornamental purposes but also serves as a rich source of bioactive essential oils. While several studies focused on the activities of the essential oils, this study analysed the potential of spent L. petersonii leaves as a natural food preservative. METHOD: We investigated the in vitro antioxidant and antimicrobial activities of crude L. petersonii extracts against activities of the purified isolated flavonoid, 6-methyltectochrysin, which was characterized using spectroscopic methods. The antioxidant assays followed ORAC, FRAP and TEAC tests. The antimicrobial activities of the extract and purified flavonoid were analysed against six multi-drug resistant microbial strains in broth dilution assays. RESULT: The results revealed that both the crude extracts and isolated 6-methyltectochrysin exhibited positive radical ion scavenging antioxidant potential, however the crude extract was about 6-fold more potent antioxidant than the purified 6-methyltectochrysin. The crude extract also showed strong antimicrobial activities against Bacillus cereus, and even more potent antimicrobial agent than the reference ampicillin antibiotic against Klebsiella pneumoniae subsp. pneumoniae. A higher resistance was observed for the tested Gram-negative strains than for the Gram-positive ones. 6-methyltectochrysin was generally inactive in the antimicrobial assays. CONCLUSION: The crude methanolic extract showed significant bioactivity which validates the medicinal relevance of the plant. The observed biological activities, especially against a notorious strain of B. cereus, suggest that L. petersonii could be a promising natural source of food preservatives.

Visualization of Aspalathin in Rooibos (Aspalathus linearis) Plant and Herbal Tea Extracts Using Thin-Layer Chromatography.

Aspalathin, the main polyphenol of rooibos (Aspalathus linearis), is associated with diverse health promoting properties of the tea. During fermentation, aspalathin is oxidized and concentrations are significantly reduced. Standardized methods for quality control of rooibos products do not investigate aspalathin, since current techniques of aspalathin detection require expensive equipment and expertise. Here, we describe a simple and fast thin-layer chromatography (TLC) method that can reproducibly visualize aspalathin in rooibos herbal tea and plant extracts at a limit of detection (LOD) equal to 178.7 ng and a limit of quantification (LOQ) equal to 541.6 ng. Aspalathin is a rare compound, so far only found in A. linearis and its (rare) sister species A. pendula. Therefore, aspalathin could serve as a marker compound for authentication and quality control of rooibos products, and the described TLC method represents a cost-effective approach for high-throughput screening of plant and herbal tea extracts.

Streptomyces swartbergensis sp. nov., a novel tyrosinase and antibiotic producing actinobacterium.

As part of an antibiotic screening program, an actinobacterium, strain HMC13T, was isolated from soil collected from the banks of the Gamka River, Western Cape Province, South Africa. The isolate was found to produce branched mycelia that differentiated into spiral spore chains with spiny spores. 16S rRNA gene sequence analysis showed the strain to be closely related to Streptomyces caelestis NRRL 2418T (99.72%) and Streptomyces azureus NBRC 12744T (99.51%). Chemotaxonomic analyses confirmed the classification of the strain as a member of the genus Streptomyces: LL-DAP in the peptidoglycan, no diagnostic sugars in the whole cell sugar pattern, dominant menaquinones including MK9(H8), MK9(H6), and the polar lipids detected included phosphatidylethanolamine. The fatty acid profile revealed the presence of mostly branched, saturated fatty acids: iso-C15:0 (14.4%), anteiso-C15:0 (21.1%), iso-C16:0 (16.8%), C16:1ω7c/2-OH iso-C15:0 (5.8%), C16:0 (6.2%), iso-C17:1ω9c (5.8%), iso-C17:0 (5.9%), and anteiso-C17:0 (9.6%). Strain HMC13T is a tyrosinase producer and exhibits very strong antibiosis against Mycobacterium aurum A+ and Staphylococcus aureus subsp. aureus ATCC 33591 (methicillin resistant), while only weak activity was observed against Bacillus cereus ATCC 10876, Enterococcus faecium VanA (vancomycin resistant), Enterococcus faecalis ATCC 51299 (vancomycin resistant) and Candida tropicalis ATCC 750T. Strain HMC13T (= LMG 28849T = NRRL B-65294T) is proposed as the type strain of a new species, to be named Streptomyces swartbergensis sp. nov.

Bacterial nitrogen fixation in sand bioreactors treating winery wastewater with a high carbon to nitrogen ratio.

Heterotrophic bacteria proliferate in organic-rich environments and systems containing sufficient essential nutrients. Nitrogen, phosphorus and potassium are the nutrients required in the highest concentrations. The ratio of carbon to nitrogen is an important consideration for wastewater bioremediation because insufficient nitrogen may result in decreased treatment efficiency. It has been shown that during the treatment of effluent from the pulp and paper industry, bacterial nitrogen fixation can supplement the nitrogen requirements of suspended growth systems. This study was conducted using physicochemical analyses and culture-dependent and -independent techniques to ascertain whether nitrogen-fixing bacteria were selected in biological sand filters used to treat synthetic winery wastewater with a high carbon to nitrogen ratio (193:1). The systems performed well, with the influent COD of 1351 mg/L being reduced by 84-89%. It was shown that the nitrogen fixing bacterial population was influenced by the presence of synthetic winery effluent in the surface layers of the biological sand filters, but not in the deeper layers. It was hypothesised that this was due to the greater availability of atmospheric nitrogen at the surface. The numbers of culture-able nitrogen-fixing bacteria, including presumptive Azotobacter spp. exhibited 1-2 log increases at the surface. The results of this study confirm that nitrogen fixation is an important mechanism to be considered during treatment of high carbon to nitrogen wastewater. If biological treatment systems can be operated to stimulate this phenomenon, it may obviate the need for nitrogen addition.

Laccase catalysis for the synthesis of bioactive compounds.

The demand for compounds of therapeutic value is increasing mainly because of new applications of bioactive compounds in medicine, pharmaceutical, agricultural, and food industries. This has necessitated the search for cost-effective methods for producing bioactive compounds and therefore the intensification of the search for enzymatic approaches in organic synthesis. Laccase is one of the enzymes that have shown encouraging potential as biocatalysts in the synthesis of bioactive compounds. Laccases are multicopper oxidases with a diverse range of catalytic activities revolving around synthesis and degradative reactions. They have attracted much attention as potential industrial catalysts in organic synthesis mainly because they are essentially green catalysts with a diverse substrate range. Their reaction only requires molecular oxygen and releases water as the only by-product. Laccase catalysis involves the abstraction of a single electron from their substrates to produce reactive radicals. The free radicals subsequently undergo homo- and hetero-coupling to form dimeric, oligomeric, polymeric, or cross-coupling products which have practical implications in organic synthesis. Consequently, there is a growing body of research focused on the synthetic applications of laccases such as organic synthesis, hair and textile dyeing, polymer synthesis, and grafting processes. This paper reviews the major advances in laccase-mediated synthesis of bioactive compounds, the mechanisms of enzymatic coupling, structure-activity relationships of synthesized compounds, and the challenges that might guide future research directions.

Effects of yeast, carboxymethylcellulose, yoghurt, transglutaminase and cyclodextrinase on mixing properties of oat dough.

The effects of yeast, carboxylmethylcellulose (CMC), plain yoghurt (YG), transglutaminase (TG) and cyclodextrinase (CG) on the mixing properties of oat dough were investigated through the use of DoughLab. A 2(5-2)fractional factorial design resolution III with yeast (1.25, 3.25 %), CMC (1, 2 %), YG (10.75, 33.75 %), TG (0.5, 1.5 %) and CG (10, 40 µl) as independent variables was implemented. The parameters measured were water absorption, arrival time, stability, energy at peak, peak resistance, development time, departure time, softening and bandwith at peak. CMC significantly (p < 0.05) increased stability, energy at peak, development and departure times, but significantly (p < 0.05) decreased water absorption, peak resistance, softening and bandwidth at peak. TG signficantly increased water absorption, peak resistance and softening, but significantly decreased energy and development time. YG significantly (p < 0.05) decreased all the parameters measured, with the exception of softening, which was significantly increased. In contrast, yeast and cyclodextrinase did not significantly affect the oat dough during mixing. Principal component analysis indicated that 85.5 % of the variation in the data could be explained by two components. Component 1 explaining 52.3 % of the variation loaded highly on dough strength (stability and departure time). Component 2 contributing 33.2 % of the variation loaded on dough resistance (water absorption and peak resistance). CMC significantly increased dough strength while yoghurt reduced it significantly. TG significantly (p < 0.05) increased the resistance of the dough to mixing while CMC and yoghurt reduced it significantly (p < 0.05). Hence, CMC, TG and yoghurt are ingredients of choice when modifying oat dough mixing properties.

Laccase applications in biofuels production: current status and future prospects.

The desire to reduce dependence on the ever diminishing fossil fuel reserves coupled with the impetus towards green energy has seen increased research in biofuels as alternative sources of energy. Lignocellulose materials are one of the most promising feedstocks for advanced biofuels production. However, their utilisation is dependent on the efficient hydrolysis of polysaccharides, which in part is dependent on cost-effective and benign pretreatment of biomass to remove or modify lignin and release or expose sugars to hydrolytic enzymes. Laccase is one of the enzymes that are being investigated not only for potential use as pretreatment agents in biofuel production, mainly as a delignifying enzyme, but also as a biotechnological tool for removal of inhibitors (mainly phenolic) of subsequent enzymatic processes. The current review discusses the major advances in the application of laccase as a potential pretreatment strategy, the underlying principles as well as directions for future research in the search for better enzyme-based technologies for biofuel production. Future perspectives could include synergy between enzymes that may be required for optimal results and the adoption of the biorefinery concept in line with the move towards the global implementation of the bioeconomy strategy.

Production of a new chitinase from Nocardiopsis halophila TN-X8 utilizing bio-waste from the blue swimming crab: enzyme characterization and immobilization.

In accordance with the framework of the Circular Blue Bioeconomy in the Mediterranean region, the objective of this study was to evaluate the biotransformation of blue swimming crab (Portunus segnis) residues obtained from the port of Sfax by an extracellular chitinase produced by Nocardiopsis halophila strain TN-X8 isolated from Chott El Jerid (Tozeur, Tunisia). From the analysis of multiple extremophilic Actinomycetota, it was determined that strain TN-X8 exclusively utilized 60 g/L of raw blue swimming crab as its carbon and energy source, achieving a chitinase activity of approximately 950 U/mL following a 6-day incubation period at 40 °C. Pure chitinase, designated as ChiA-Nh30, was obtained after heat treatment, followed by ammonium sulfate fractionation and Sephacryl® S-200 column chromatography. The maximum ChiA-Nh30 activity was observed at pH 3 and 75 °C. Interestingly, compared with cyclohexamidine, ChiA-Nh30 showed a good antifungal effect against four pathogenic fungi. Furthermore, when using colloidal chitin as substrate, ChiA-Nh30 demonstrated a higher degree of catalytic efficiency than the commercially available Chitodextrinase®. In addition, ChiA-Nh30 could be immobilized by applying encapsulation and encapsulation-adsorption techniques. The kaolin and charcoal used acted as excellent binders, resulting in improved ChiA-Nh30 stability. For the immobilized ChiA-Nh30, the yield of N-acetyl-D-glucosamine monomers released from 20% (w/v) blue swimming crab residues increased by 3.1 (kaolin) and 2.65 (charcoal) times, respectively.

Amycolatopsis umgeniensis sp. nov., isolated from soil from the banks of the Umgeni River in South Africa.

A novel member of the genus Amycolatopsis was isolated from soil collected from the banks of the Umgeni River, KwaZulu Natal province, South Africa. The strain, designated UM16(T), grouped with the type strains of Amycolatopsis alba, Amycolatopsis coloradensis and Amycolatopsis thailandensis by 16S rRNA gene based phylogeny. Genetic distance values, based on the gyrB and recN genes, between strain UM16(T) and its closest relatives were all above the threshold values of 0.02 and 0.04, respectively, that have been proposed to distinguish Amycolatopsis type strains. DNA-DNA hybridisation experiments confirmed that strain UM16(T) represents a unique genomic species, sharing 18.4 ± 5.1, 16.2 ± 1.8 and 45.8 ± 8.9 % DNA relatedness to the type strains of A. alba, A. coloradensis and A. thailandensis, respectively. The physiological, phylogenetic and DNA-relatedness data support the description of strain UM16(T) as the type strain of a novel species, for which the name Amycolatopsis umgeniensis sp. nov. is proposed (= DSM 45272(T) = NRRL B-24724(T)).

Bioactive components in Bambara groundnut (Vigna subterraenea (L.) Verdc) as a potential source of nutraceutical ingredients.

The utilization of nutraceuticals on a global scale has significantly increased over the past few years due to their reported health benefits and consumer's reluctance to consume synthetic drugs. This paper provides information regarding new and potential value added uses of biologically active compounds in Bambara groundnut (BGN) as ingredients that could be further researched and exploited for various applications. Nutraceutical is a food or part of food that apart from providing basic nutrients, offers medicinal benefits either by prevention and or treatment of an illness. BGN is a legume with rich nutrient profile that is under exploited industrially. It is widely used in African traditional medicine for its various health outcome, but has not been explored scientifically for its numerous nutraceutical potentials. Compared to beans BGN has greater quantity of soluble fiber and also have high dietary fiber. It is rich in polyphenolic compound which include flavonoids subgroups like flavonols, flavanols, anthocyanindins, isoflavones and phenolic acids: both benzoic acid and cinnamic acid derivatives, biologically active polyunsaturated fatty acids, proteins and peptides, antioxidant vitamins and minerals. The rising interest and emphasis in plant-based biologically active components (nutraceuticals) for various health promotion, has positioned this African legume as a potential source of nutraceutical ingredients (bioactive components) that could be exploited for improved nutrition and health.

Impact of Physicochemical Parameters on the Diversity and Distribution of Microbial Communities Associated with Three South African Peatlands.

Peatlands are complex wetland-like ecosystems that harbor diverse microbial communities. In this study, the microbial communities (fungal and actinobacterial) associated with an unimpacted peatland (Vankervelsvlei; VV), an impacted peatland (Goukou River system; GK), and a developing peatland (Nuwejaars River system; NR) were determined through ITS and 16S rRNA metataxonomic analyses. Unidentified Acidimicrobiales dominated in GK and NR, unidentified Intrasporangiaceae and Solirubobacterales in NR, and Corynebacterium, Propionibacterium, and Streptomyces species in VV. The fungal phyla, Ascomycota and Basidiomycota, dominated all three sites, and harbored unique fungal taxa belonging to a wide range of fungal guilds. Physicochemical properties of the peat collected from the three sites were analyzed in association with microbial community structures in order to determine which parameters acted as the main drivers for microbial diversity. BEST analysis (linking microbial diversity patterns to environmental variables) showed that nitrogen (N), aluminum (Al), phosphorus (P), and potassium (K) were the most significant physicochemical drivers of actinobacterial community structure, while iron (Fe) and humification were the environmental parameters that affected the fungal communities the most. In conclusion, this study has provided some insight into the fungal and actinobacterial communities associated with three South African peatlands and the main environmental drivers that influence these communities.

Preparation, characterization, immobilization, and molecular docking analysis of a novel detergent-stable subtilisin-like serine protease from Streptomyces mutabilis strain TN-X30.

We recently described the production of a detergent-biocompatible crude protease from Streptomyces mutabilis strain TN-X30. Here, we describe the purification, characterization, and immobilization of the serine alkaline protease (named SPSM), as well as the cloning, sequencing, and over-expression of its corresponding gene (spSM). Pure enzyme was obtained after ammonium sulphate precipitation followed by heat-treatment and Sephacryl® S-200 column purification. The sequence of the first 26 NH2-terminal residues of SPSM showed a high sequence identity to subtilisin-like serine proteases produced by actinobacteria. The spSM gene was heterologously expressed in Escherichia coli BL21(DE3)pLysS and E. coli BL21-AI™ strains using pTrc99A (rSPSM) and Gateway™ pDEST™ 17 [(His)6-tagged SPSM] vectors, respectively. Results obtained indicated that the (His)6-tagged SPSM showed the highest stability. The SPSM was immobilized using encapsulation and adsorption-encapsulation approaches and three different carriers. Features of SPSM in soluble and immobilized forms were analyzed by Fourier transform infrared (FTIR) spectroscopy in attenuated total reflection (ATR) mode, X-ray diffraction (XRD), zeta potential measurements, and field emission scanning electron microscopy (FE-SEM). The white clay and kaolin used in this study are eco-friendly binders to alginate-SPSM and show great potential for application of the immobilized SPSM in various industries. Molecular modeling and docking of N-succinyl-l-Phe-l-Ala-l-Ala-l-Phe-p-nitroanilide in the active site of SPSM revealed the involvement of 21 amino acids in substrate binding.

Actinobacteria isolated from termite guts as a source of novel oxidative enzymes.

A multi-faceted screening programme was designed to search for the oxidases, laccase, peroxidase and tyrosinase. Actinobacteria were selectively isolated from the paunch and colon region of the hindguts of the higher termite, Amitermes hastatus. The isolates were subjected to solid media assays (dye decolourization, melanin production and the utilization of indulin AT as sole carbon source) and liquid media assays. Eleven of the 39 strains had the ability to decolourize the dye RBBR, an indicator for the production of peroxidases in actinobacteria. Melanin production on ISP6 and ISP7 agar plates served as a good indicator for laccase and/or tyrosinase production and the ability of the strains to grow in the presence of indulin AT as a sole carbon source served as a good indicator of lignin peroxidase and/or general peroxidase production. Enzyme-producing strains were cultivated in liquid media and extracellular enzyme activities measured. Strains with the ability to produce oxidative enzymes under the conditions tested were identified to genus level by 16S rRNA gene analysis and compared to known oxidase producers. A strong relationship was observed between the environment sampled (termite guts where lignocellulose degradation occurs) and the dominant type of oxidative enzyme activity detected (laccases and peroxidases), which suggests the possibility of future targeted screening protocols linking the physical properties of the target enzymes with specific operational conditions required, such as lignocellulosic degradation in the preparation of biofuel feedstocks.

Effects of Clonostachys rosea f. catenula Inoculum on the Composting of Cabbage Wastes and the Endophytic Activities of the Composted Material on Tomatoes and Red Spider Mite Infestation.

Globally, fungal inocula are being explored as agents for the optimization of composting processes. This research primarily evaluates the effects of inoculating organic vegetable heaps with the entomopathogenic fungus Clonostachys rosea f. catenula (Hypocreales) on the biophysicochemical properties of the end-product of composting. Six heaps of fresh cabbage (Brassica oleracea var. capitata) waste were inoculated with C. rosea f. catenula conidia and another six were not exposed to the fungus. The composted materials from the fungus- and control-treated heaps were subsequently used as a medium to cultivate tomatoes (Solanum lycopersicum). The biophysicochemical characteristics of the composted materials were also assessed after composting. In addition, the protective effect of the fungal inoculum against red spider mite (Tetranychus urticae) infestations in the tomatoes was evaluated through the determination of conidial colonization of the plant tissue and the number of plants infested by the insect. Furthermore, phytotoxicity tests were carried out post experiment. There were few significant variations (p < 0.05) in heap temperature or moisture level between treatments based on the weekly data. We found no significant differences in the levels of compost macronutrient and micronutrient constituents. Remarkably, the composted materials, when incorporated into a growth medium from fungus-treated heaps, induced a 100% endophytic tissue colonization in cultivated tomato plants. While fewer red spider mite infestations were observed in tomato plants grown in composted materials from fungus-treated heaps, the difference was not significant (χ2 = 0.96 and p = 0.32). The fungal treatment yielded composted materials that significantly (p < 0.05) enhanced tomato seed germination, and based on the phytotoxicity test, the composted samples from the heaps exposed to the C. rosea f. catenula inoculum were not toxic to tomato seeds and seedlings. In conclusion, this study showed that C. rosea f. catenula improved the quality of composted materials in terms of fungal endophytism and seed germination.