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Paper spray mass spectrometry (PS-MS) is an ambient ionization technique that allows for rapid and direct mass spectrometry analysis for a wide range of chemical compounds due to its portability, little to no sample preparation, and cost-effective materials. As applications with this technique continue to expand, the identification and discrimination of bacteria at the strain level remain a promising avenue for researchers. Although studies in the past demonstrated the applicability of PS-MS to discriminate bacteria at the strain level, no one has reported the strain-level differentiation of actinobacteria without using solvent for PS-MS. Hence, this study demonstrates that optimization of PS-MS permits the investigation and differentiation of the metabolic profiles of actinobacteria without the need for solvents, diminishing the potential for sample contamination and consequently increasing the versatility of this technique. In doing so, strains of actinobacteria (CAAT P5-21, CAAT P5-16, CAAT 8-25, CAAT P8-92, and CAAT P11-13) were grown and transferred to produce a crude growth medium. The supernatant was used for the PS-MS analyses using a Thermo Scientific LTQ mass spectrometer. Multivariate statistical analysis, including principal component analysis (PCA) and hierarchal cluster analysis (HCA), was employed to chemically distinguish the strains of bacteria. As a result, each strain of actinobacteria could be visually differentiated based on their metabolic profile. These findings demonstrate the practicability of using a liquid medium as an alternative to many other organic solvents when analyzing bacteria, making PS-MS a crucial addition to a microbiologist's research toolkit., (© 2023 John Wiley & Sons Ltd.)

Designing novel isolation methods could enhance the diversification of the available bacterial strains to biotechnology. In this study, the new ex situ and in situ cultivation methods are introduced for the isolation of actinobacteria. In the ex situ experiments, the soil suspension was spread on the isolation media located above some ordinary papers in immediate contact with the slurry of soil substrate and incubated for 16 weeks. The paper was wholly immersed in the cave soil for in situ cultivations, and the containers were buried under layers of soil in Hampoeil cave for 10 weeks. Fruit wrapping kraft coated paper, with 68.8% recovery of isolates, was a better choice in isolation of actinobacteria than other studied filter paper. Based on the molecular identification results, 19% of the isolates obtained from the in situ cultivation method had less than 98.5% similarity to known taxa of actinobacteria and potentially may represent new species. In contrast, in the standard cultivation method, 1.3% of the isolates had less than 98.5% similarity 16Sr RNA gene. This data shows that the introduced cultivation method is a promising technique for isolating less culturable or new actinobacteria., (© 2021. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)

Composting is an alternative method to dispose of de-inking paper sludge (DPS). Today, few studies have investigated the water-soluble carbon (WSC) substances as indicators of the decomposition process and the microbial changes taking place during the composting of DPS. Accordingly, the goal is to study their dynamics during the composting of DPS at three nitrogen levels, 0.6%, 0.7% or 0.9% total N, using mechanical turning. The changes in WSC substances, microbial biomass carbon (MBC) and, total and DPS microbial populations were monitored during 24 weeks. Also, microorganisms were identified and tested for the production of selected enzymes. Regardless of N treatments, the dynamic of WSC substances indicated that cellulose and hemicellulose fractions of DPS fibers were mainly biodegraded during the first 8 weeks while the more resistant carbon (C) fractions were biodegraded thereafter. MBC also evolved regardless of N treatments but was correlated to WSC substances. Its high values decline mostly after 12 weeks indicating the exhaustion of this source of C energy for microbial growth and the stabilisation of DPS organic matter. The dynamic and identified microorganisms were comparable to those observed in other composting processes. However, the results pointed out that those mostly implicated in the hydrolysis of DPS fibers were the thermophilic actinomycetes and fungi and, by comparison to the 0.6% or 0.7% N treatment, they decreased in presence of the 0.9% N treatment. Most microorganisms were hemicellulolytic bacteria, while actinomycetes and fungi were capable of degrading a wide variety of substrates. Overall, dynamics of WSC substances and microbial populations indicated that during composting, DPS decomposition obey a two phase decay while, contrary to the lowest N treatment, the 0.9% N treatment has slowed down this process by harming the important microbial populations implicated in the degradation of DPS fibers.

Actinomycetes-a group of antimicrobial producing bacteria-have been successfully cultured and characterized from the nest material of diverse arthropods. Some are symbionts that produce antimicrobial chemicals found to protect nest brood and resources from pathogenic microbes. Others have no known fitness relationship with their associated insects, but have been found to produce antimicrobials in vitro. Consequently, insect nest material is being investigated as a new source of novel antimicrobial producing actinomycetes, which could be harnessed for therapeutic potential. To extend studies of actinomycete-insect associations beyond soil-substrate dwelling insects and wood boring excavators, we conducted a preliminary assessment of the actinomycetes within the nests of the paper wasp, Polistes dominulus (Christ). We found that actinomycetes were readily cultured from nest material across multiple invasive P. dominulus populations-including members of the genera Streptomyces, Micromonospora, and Actinoplanes. Thirty of these isolates were assayed for antimicrobial activity against the challenge bacteria Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Serratia marcescens, and Bacillus subtilis. Sixty percent of isolates inhibited the growth of at least one challenge strain. This study provides the first assessment of bacteria associated with nests of P. dominulus, and the first record of antimicrobial producing actinomycetes isolated from social wasps. We provide a new system to explore nest associated actinomycetes from a ubiquitous and cosmopolitan group of insects.

Aims: The utilization of micro‐organisms in pulp and paper industries has proved biobleaching technology as an environmentally friendly alternative to the conventional approach. In this paper, the effect of actinobacterial fermentation broth on pulp biobleaching has been investigated. Methods and Results: Actinobacterial colonies were isolated from lignocellulose‐rich soil samples and screened for xylanase production and bleaching activity. The most efficient isolate in bleaching activity showed 100% similarity to Streptomyces rutgersensis. Pulp treatment with 5‐day fermentation broth of this strain showed up to 7% increase in brightness (30°C for 6 h, pH (5–7)) compared to untreated (control) pulp. Also, after 60 min biotreatment, significant reduction (12·5%) in consumption of bleaching chemicals was achieved to obtain final brightness of 55%. Conclusion: Actinobacterial fermentation broth can be considered as a rich source of effective biobleaching agents which may be considered as environmental friendly and cost‐effective technique in comparison with traditional method. Significance and Impact of the Study: Our findings showed ability of S. rutgersensis UTMC 2445 in bleaching chemomechanical paper pulp. Also, two strains of Saccharothrix, a rare actinobacterium, with biobleaching activity were introduced. In the proposed method, there is no need to use purified enzymes, and biobleaching process can be done using the fermentation broth. [ABSTRACT FROM AUTHOR]

Actinomycetes are a suitable microbial group for the synthesis of lignocellulose-degrading enzymes. Enzymes that may degrade organic material, including cellulose, hemicellulose, and lignin, are released by actinomycetes. The aim of this research was to isolate actinomycetes from Rajkot, Gujarat, India’s soil and evaluate the activity of their cellulase and Xylanase enzymes. Starch Casein Agar (SCA) was used to identify a total of 30 isolates of actinomycetes. A qualitative plate assay (CMC-Na, Congo red) revealed that the highest zone of catalysis for MMD1 was 36 mm. Five strains were discovered to be effective for quantitative quantification of endoglucanase utilising filter paper and CMC as substrates: MMD1, MMD2, MMD3, MMD4, and MMD8. Following MMD 1 (endoglucanase 5.4 IU; FPase 4.4 IU), MMD 2 (endoglucanase 4.5 IU; FPase 3.4 IU) has demonstrated considerable endoglucanase and FPase activity. Beechwood xylan was used to treat sugarcane bagasse in order to test Xylanase, and 45% of the xylan (hemicellulose) fraction was obtained. MMD1 and MMD2 measured the xylanase enzyme activity (4.8IU and 4.2IU) in quantitative and qualitative assays (34 mm and 22 mm for BWX and 32 mm and 14 mm for agro-waste xylan). The strain MMD1 was identified as Streptomyces chartreusis through morphological, biochemical, and finally molecular characterization by 16S rRNA sequencing. It was then submitted to NCBI GenBank with the accession number MT254830. [ABSTRACT FROM AUTHOR]

The fungal system holds morphological plasticity and metabolic versatility which makes it unique. Fungal habitat ranges from the Arctic region to the fertile mainland, including tropical rainforests, and temperate deserts. They possess a wide range of lifestyles behaving as saprophytic, parasitic, opportunistic, and obligate symbionts. These eukaryotic microbes can survive any living condition and adapt to behave as extremophiles, mesophiles, thermophiles, or even psychrophile organisms. This behaviour has been exploited to yield microbial enzymes which can survive in extreme environments. The cost-effective production, stable catalytic behaviour and ease of genetic manipulation make them prominent sources of several industrially important enzymes. Pectinases are a class of pectin-degrading enzymes that show different mechanisms and substrate specificities to release end products. The pectinase family of enzymes is produced by microbial sources such as bacteria, fungi, actinomycetes, plants, and animals. Fungal pectinases having high specificity for natural sources and higher stabilities and catalytic activities make them promising green catalysts for industrial applications. Pectinases from different microbial sources have been investigated for their industrial applications. However, their relevance in the food and textile industries is remarkable and has been extensively studied. The focus of this review is to provide comprehensive information on the current findings on fungal pectinases targeting diverse sources of fungal strains, their production by fermentation techniques, and a summary of purification strategies. Studies on pectinases regarding innovations comprising bioreactor-based production, immobilization of pectinases, in silico and expression studies, directed evolution, and omics-driven approaches specifically by fungal microbiota have been summarized., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Thermophilic actinomycetes are commonly found in extreme environments and can thrive and adapt to extreme conditions. These organisms exhibit substantial variation and garnered significant interest due to their remarkable enzymatic activities. This study evaluated the potential of Streptomyces griseorubens NBR14 and Nocardiopsis synnemataformans NBRM9 strains to produce thermo-stable amylase via submerged fermentation using wheat and bean straw. The Box-Behnken design was utilized to determine the optimum parameters for amylase biosynthesis. Subsequently, amylase underwent partial purification and characterization. Furthermore, the obtained hydrolysate was applied for ethanol fermentation using Saccharomyces cerevisiae. The optimal parameters for obtaining the highest amylase activity by NBR14 (7.72 U/mL) and NBRM9 (26.54 U/mL) strains were found to be 40 and 30 °C, pH values of 7, incubation time of 7 days, and substrate concentration (3 and 2 g/100 mL), respectively. The NBR14 and NBRM9 amylase were partially purified, resulting in specific activities of 251.15 and 144.84 U/mg, as well as purification factors of 3.91 and 2.69-fold, respectively. After partial purification, the amylase extracted from NBR14 and NBRM9 showed the highest activity level at pH values of 9 and 7 and temperatures of 50 and 60 °C, respectively. The findings also indicated that the maximum velocity (V max ) for NBR14 and NBRM9 amylase were 57.80 and 59.88 U/mL, respectively, with K m constants of 1.39 and 1.479 mM. After 48 h, bioethanol was produced at concentrations of 5.95 mg/mL and 9.29 mg/mL from hydrolyzed wheat and bean straw, respectively, through fermentation with S. cerevisiae. Thermophilic actinomycetes and their α-amylase yield demonstrated promising potential for sustainable bio-ethanol production from agro-byproducts., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Microbial degradation of lignin, a natural complex biopolymer, a renewable raw material with a wide range of applications, has been mainly directed at fungal systems, nevertheless, recent studies have proposed the bacterial role in lignin degradation and modification since bacteria possess remarkable environmental adaptability, and various production of enzymes and biochemistry. An occurrence of a high proportion of lignin-degrading genes has been confirmed in actinobacteria and proteobacteria classes by bioinformatics analysis, which points to the probability of undiscovered pathways and enzymes. Because of that, bacterial lignin decomposition might be substantially different from fungal lignin decomposition. Bacteria capable of lignin modification and degradation belong to actinomycetes, some Firmicutes, α-proteobacteria, and γ-proteobacteria. The enzymes responsible for lignin degradation are lignin peroxidase, manganese-dependent peroxidase, versatile peroxidase, dye-decolourizing peroxidase, and laccases. One of the main lignin producers is the pulp and paper manufacturing industry. Lignolytic microorganisms have been identified from diverse habitats, such as in plants, soil, wood, and the gut. Bacterial strains Bacillus, Rhodococcus, Sterptomyces, and Pseudomonas have been reported to have lignin decomposition ability. This review aims to describe the role of bacteria in lignin degradation, bacterial species, and bacterial enzymes included in lignin degradation. Several reports about bacterial species involved in lignin degradation are also highlighted, and the current state of the knowledge on the degradation of lignin from the pulp and paper manufacturing industry are reported. [ABSTRACT FROM AUTHOR]

In this study, we employed a polyphasic approach to determine the taxonomic position of a newly isolated actinomycete, designated SE31 T , obtained from a sediment sample collected at Cape Rochado, Malaysia. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain SE31 T belonged to the family Pseudonocardiaceae and exhibited the highest sequence similarity (98.9%) to Sciscionella marina. Further genomic analysis demonstrated a 93.4% average nucleotide identity and 54.4% digital DNA-DNA hybridization relatedness between strain SE31 T and S. marina. The chemotaxonomic characteristics of strain SE31 T were typical of the genus Sciscionella, including cell-wall chemotype IV (with meso-diaminopimelic acid as the diagnostic diamino acid, and arabinose and galactose as whole-cell sugars). The identified polar lipids of strain SE31 T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, and hydroxyphosphatidymethylethanolamine. The primary menaquinone observed was MK-9(H 4 ), and the major cellular fatty acid was iso-C 16:0 . The genomic DNA size of strain SE31 T was determined to be 7.4 Mbp with a G+C content of 68.7%. Based on these comprehensive findings, strain SE31 T represents a novel species within the genus Sciscionella, in which the name Sciscionella sediminilitoris sp. nov. is proposed. The type strain of Sciscionella sediminilitoris is SE31 T (= DSM 46824 T  = TBRC 5134 T )., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Most biosynthetic gene clusters (BGCs) of Actinobacteria are either silent or expressed less than the detectable level. The non-genetic approaches including biological interactions, chemical agents, and physical stresses that can be used to awaken silenced pathways are compared in this paper. These non-genetic induction strategies often need screening approaches, including one strain many compounds (OSMAC), reporter-guided mutant selection, and high throughput elicitor screening (HiTES) have been developed. Different types of genetic manipulations applied in the induction of cryptic BGCs of Actinobacteria can be categorized as genome-wide pleiotropic and targeted approaches like manipulation of global regulatory systems, modulation of regulatory genes, ribosome and engineering of RNA polymerase or phosphopantheteine transferases. Targeted approaches including genome editing by CRISPR, mutation in transcription factors and modification of BGCs promoters, inactivation of the highly expressed biosynthetic pathways, deleting the suppressors or awakening the activators, heterologous expression, or refactoring of gene clusters can be applied for activation of pathways which are predicted to synthesize new bioactive structures in genome mining studies of Acinobacteria. In this review, the challenges and advantages of employing these approaches in induction of Actinobacteria BGCs are discussed. Further, novel natural products needed as drug for pharmaceutical industry or as biofertilizers in agricultural industry can be discovered even from known species of Actinobactera by the innovative approaches of metabolite biosynthesis elicitation., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The aim of this study was to characterize the actinobacterial population present in pulps and boards containing recycled fibres. A total of 107 isolates was identified on the basis of their pigmentation, morphological properties, fatty acid profiles and growth temperature. Of the wet pulp and water sample isolates (n=87), 74.7% belonged to the genus Streptomyces, 17.2% to Nocardiopsis and 8.0% to thermoactinomycetes, whereas all the board sample isolates (n=20) were thermoactinomycetes. The identification of 53 isolates was continued by molecular methods. Partial 16S rDNA sequencing and automated ribotyping divided the Streptomyces isolates (n=31) into 14 different taxa. The most common streptomycetes were the mesophilic S. albidoflavus and moderately thermophilic S. thermocarboxydus. The Nocardiopsis isolates (n=11) belonged to six different taxa, whereas the thermoactinomycetes were mainly members of the species Laceyella sacchari (formerly Thermoactinomyces sacchari). The results indicated the probable presence of one or more new species within each of these genera. Obviously, the drying stage used in the board making processes had eliminated all members of the species Streptomyces and Nocardiopsis present in the wet recycled fibre pulp samples. Only the thermotolerant endospores of L. sacchari were still present in the final products. The potential of automated ribotyping for identifying actinobacteria was indicated, as soon as comprehensive identification libraries became available.

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., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Harmful cyanobacterial blooms that occur in freshwater can produce hazardous cyanotoxins as contaminants that threaten ecosystems, aquatic organisms, and human health. In the present study, the actinobacterium Streptomyces enissocaesilis, strain M35, isolated from soils, exhibited the strongest algicidal activity against the toxic cyanobacterium Phormidium angustissimum TISTR 8247. To improve the P. angustissimum removal efficiency of strain M35, the optimum carbon and nitrogen sources were determined as starch and yeast extract, respectively. Response surface methodology (RSM) using the Box-Behnken design (BBD) revealed that the optimal independent parameters among the culture medium conditions for enhancing the algicidal activity of strain M35 were 21.5 g/L starch, 0.57 g/L yeast extract, and a pH value of 8.00. The Phormidium sp. removal efficiency increased notably from 80.8 to 94.4% under the optimum conditions. In a batch experiment, the removal of P. angustissimum in an internal airlift loop (IAL) bioreactor containing immobilized strain M35 on a plastic medium indicated a high anti-Phormidium activity of 94.8%, whereas in a continuous system, strain M35 exhibited a removal efficiency of 85.5%. This study revealed that this actinobacterium could potentially be utilized to remove the toxic cyanobacterium Phormidium from water., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Indigenous microbial diversity has potential for rapid decomposition of residue through enzyme activities that is alternative, effective, and environment friendly strategy to accelerate degradation of lignocellulose in agricultural residues and make composting process economically viable. Keeping this view, the main objective of the present study was isolation and characterization of lignocellulosic degrading microbial diversity from long-term residue management practice experiments and to develop potential microbial consortium for rapid degradation of lignocellulosic biomass. In this study, twenty-five bacteria, nine fungi, and four actinomycetes isolates were obtained from the soil samples of different residue management fields from Ludhiana, Punjab, India. All isolates were qualitatively and quantitatively screened for enzyme activities, i.e., cellulase, xylanase, laccase, and lignin peroxidase. On the basis of quantitative estimation of enzyme activities, 3 fungal (S1F1, S2F4, and S6F9), 2 actinomycetes (S1A1 and S6A4), and 2 bacterial strains (S6B16 and S6B17) were further selected for in vitro bio-compatibility assay. Selected bio-compatible microbial strains were identified as Streptomyces flavomacrosporus (S6A4), Aspergillus terreus (S2F4), and Bacillus altitudinis (S6B16) through 16S rRNA and 18S rRNA sequencing. Furthermore, single and developed microbial consortium (S6B16 + S6A4 + S2F4) were screened for quantitative estimation of cellulase, xylanase, laccase, and lignin peroxidase enzymes with 23 biochemically different cereal, legume, and oil seed crop residues for optimization of enzyme activities at different time intervals. Results revealed that Vigna radiata followed by Cajanus cajan and Arachis hypogaea straw residue powder @ 1% in culture broth are a promising carbon source for B. altitudinis, S. flavomacrosporus, and A. terreus to produce higher ligno-cellulolytic microbial degrading enzymes due to variable range of carbon (C):nitrogen (N) ratio and higher ligno-cellulolytic content in the studied crop residues. Thus, the application of indigenous microbial consortium with efficient lignocellulose hydrolysis enzyme machinery might be an attractive alternative for ex situ crop residue management practices under sustainable manners., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

India has the highest burden of Tuberculosis (TB) and multidrug-resistant TB (MDR-TB) worldwide. Innovative technology is the need of the hour to identify these cases that remain either undiagnosed or inadequately diagnosed due to the unavailability of appropriate tools at primary healthcare settings. We developed and evaluated 3 kits, namely ‘TB Detect’ (containing BioFM-Filter device), ‘TB Concentration and Transport’ (containing Trans-Filter device) and ‘TB DNA Extraction’ kits. These kits enable bio-safe equipment-free concentration of sputum on filters and improved fluorescence microscopy at primary healthcare centres, ambient temperature transport of dried inactivated sputum filters to central laboratories and molecular detection of drug resistance by PCR and DNA sequencing (Mol-DST). In a 2-site evaluation (n = 1190 sputum specimens) on presumptive TB patients, BioFM-Filter smear exhibited a significant increase in positivity of 7% and 4% over ZN smear and LED-FM smear (p<0.05), respectively and an increment in smear grade status (1+ or 2+ to 3+) of 16% over ZN smear and 20% over LED-FM smear. The sensitivity of Mol-DST in presumptive MDR-TB and XDR-TB cases (n = 148) was 90% for Rifampicin (95% confidence interval [CI], 78–96%), 84% for Isoniazid (95% CI, 72–92%), 83% for Fluoroquinolones (95% CI, 66–93%) and 75% for Aminoglycosides (95% CI, 35–97%), using phenotypic DST as the reference standard. Test specificity was 88–93% and concordance was ~89–92% (κ value 0.8–0.9). The patient-friendly kits described here address several of the existing challenges and are designed to provide ‘Universal Access’ to rapid TB diagnosis, including drug-resistant disease. Their utility was demonstrated by application to sputum at 2 sites in India. Our findings pave the way for larger studies in different point-of-care settings, including high-density urban areas and remote geographical locations. [ABSTRACT FROM AUTHOR]

Actinobacteria constitute a highly diverse bacterial phylum with an unrivalled metabolic versatility. They produce most of the clinically used antibiotics and a plethora of other natural products with medical or agricultural applications. Modern 'omics'-based technologies have revealed that the genomic potential of Actinobacteria greatly outmatches the known chemical space. In this Review, we argue that combining insights into actinobacterial ecology with state-of-the-art computational approaches holds great promise to unlock this unexplored reservoir of actinobacterial metabolism. This enables the identification of small molecules and other stimuli that elicit the induction of poorly expressed biosynthetic gene clusters, which should help reinvigorate screening efforts for their precious bioactive natural products.

Cyanogramide (1) from the marine actinomycete Actinoalloteichus cyanogriseus WH1-2216-6 features a unique spirooxindole skeleton and exhibits significant bioactivity to efficiently reverse drug resistance in tumor cells. The biosynthetic gene cluster of 1 in A. cyanogriseus WH1-2216-6 was identified and refactored by promoter engineering for heterologous expression in Streptomyces coelicolor YF11, thereby enabling the production of 1 and five new derivatives. Interesting, four of them, including 1, were identified as enantiomeric mixtures in different ratios. The functions of tailoring enzymes, including two methyltransferases (CyaEF), and three cytochrome P450 monooxygenases (CyaGHI) were confirmed by gene inactivation and feeding experiments, leading to the elucidation of a concise biosynthetic pathway for 1. Notably, CyaH was biochemically verified to catalyze the formation of the spirooxindole skeleton in 1 through an unusual carbocation-mediated semipinacol-type rearrangement reaction., (© 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Bacteria are an enormous and largely untapped reservoir of biosensing proteins. We describe an approach to identify and isolate bacterial allosteric transcription factors (aTFs) that recognize a target analyte and to develop these TFs into biosensor devices. Our approach utilizes a combination of genomic screens and functional assays to identify and isolate biosensing TFs, and a quantum-dot Förster Resonance Energy Transfer (FRET) strategy for transducing analyte recognition into real-time quantitative measurements. We use this approach to identify a progesterone-sensing bacterial aTF and to develop this TF into an optical sensor for progesterone. The sensor detects progesterone in artificial urine with sufficient sensitivity and specificity for clinical use, while being compatible with an inexpensive and portable electronic reader for point-of-care applications. Our results provide proof-of-concept for a paradigm of microbially-derived biosensors adaptable to inexpensive, real-time sensor devices.

The search for microorganisms with potential for bioconversion of lignocellulose is now of immediate interest. Industrial waste is a source of various microorganisms. This paper describes results of the research of potentially lignocellulolytic actinobacteria isolated from activated sludge of the wastewater treatment plant of a pulp and paper mill located in Komi Republic (Russia). One strain of actinobacteria, AI2, was found to be sufficiently active in terms of degradation of lignocellulose-containing materials. Testing of the AI2 isolate demonstrated its ability to synthesize cellulase, dehydrogenase and protease to various extents. The AI2 strain was found capable of biosynthesizing cellulase to 5.5 U/ml. In case of solid-phase fermentation using treated softwood and hardwood sawdust, the content of main components changed most significantly in aspen sawdust: from initial concentration of 20.4% down to 15.6% for lignin, and from 50.6% down to 31.8% for cellulose. In case of liquid-phase fermentation, the content of lignin components decreased significantly in the treated aqueous medium that contained lignosulfonates: from initial concentration of 3.6 g down to 2.1g. Taxonomic study of the AI2 strain of actinobacteria confirmed that it belongs to the rare Pseudonocardia genus of actinomycetes. Based on the results of 16S rRNA sequencing, the AI2 strain is most similar to the species Pseudonocardia carboxydivorans. [ABSTRACT FROM AUTHOR]

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is the first-line method for the rapid identification of most cultured microorganisms. As for Streptomyces strains, MALDI-TOF MS identification is complicated by the characteristic incrustation of colonies in agar and the strong cell wall of Actinomycetes cells requiring the use of alternative protein extraction protocols. In this study, we developed a specific protocol to overcome these difficulties for the MALDI-TOF MS identification of Actinomycetes made on solid medium. This protocol includes incubation of colony removed from agar plate with the beta-agarase enzyme, followed by a mechanical lysis and two washes by phosphate buffer and ethanol. Twenty-four Streptomyces and two Lentzea strains isolated from Algerian desertic soils were first identified by 16S rRNA sequencing as gold standard method, rpoB gene was used as a secondary gene target when 16S rRNA did not allow species identification. In parallel the isolates were identified by using the MALDI-TOF MS protocol as reported. After the expansion of the database with the inclusion of this MSP S , the strains were analyzed again in MALDI Biotyper, and all were identified. This work demonstrates that the rapid identification of Actinomycetes can be obtained without protein extraction step frequently used in MALDI-TOF mass spectrometry with this type of microorganisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Natural products (NPs) produced by bacteria, particularly soil actinomycetes, often possess diverse bioactivities and play a crucial role in human health, agriculture, and biotechnology. Soil actinomycete genomes contain a vast number of predicted biosynthetic gene clusters (BGCs) yet to be exploited. Understanding the factors governing NP production in an ecological context and activating cryptic and silent BGCs in soil actinomycetes will provide researchers with a wealth of molecules with potential novel applications. Here, we highlight recent advances in NP discovery strategies employing ecology-inspired approaches and discuss the importance of understanding the environmental signals responsible for activation of NP production, particularly in a soil microbial community context, as well as the challenges that remain., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Large amount of sulphur is released by the combustion of fossil fuels in the form of So X which affects human health and leads to acid rain. To overcome this issue, it is essential to eliminate sulphur moieties from heterocyclic organo-sulphur compounds like Dibenzothiophene (DBT) present in the petrol. In this study Surface enhanced Raman scattering (SERS) spectroscopy is used to analyze the desulfurizing activity of Tsukamurella paurometabola bacterial strain. The most prominent SERS peaks observed at 791, 837, 944 and 1032 cm -1 , associated to C-S stretching, are solely observed in dibenzothiophene and its metabolite-I (DBTS) but absent in 2-Hydroxybiphenyl (metabolite-II) and extraction sample of supernatant as a result of biodesulfurization. Moreover, the SERS peaks observed at 974 (characteristic peak of benzene ring) and 1015 cm -1 is associated to C-C ring breathing while 1642 and 1655 cm -1 assigned to CC bonds of aromatic ring. These peaks are only observed in 2-Hydroxybiphenyl (metabolite-II) and extraction sample of supernatant as a result of biodesulfurization. Notably, these peaks are absent in the Dibenzothiophene and its metabolite-I which indicate that aromatic ring is carrying sulfur in this fraction. Moreover, multivariate data analytical tools like principal component analysis (PCA) and PCA-loadings are applied to further differentiate between dibenzothiophene and its metabolites that are Dibenzothiophene sulphone (metabolite-I) and 2-Hydroxybiphenyl (metabolite-II)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Two novel actinobacteria, designated as SYSU M7M538 T and SYSU M7M531, were isolated from oral of Eumetopias jubatus in Zhuhai Chimelong Ocean Kingdom, China. The cells of these microorganisms stained Gram-positive and were rod shaped. These strains were facultative anaerobic, and catalase-positive. Optimal growth occurred at 37 °C and pH 7.0 over 7 days of cultivation. Both strains possessed diphosphatidylglycerol, phosphatidylglycerol and phosphocholine as the major polar lipids. The main menaquinone was MK-9(H 4 ). The major fatty acids were C 16:0 , C 17:1 w 8 c , C 17:0 , C 18:1 w 9 c and C 18:0 . Analyses of genome sequences revealed that the genome size of SYSU M7M538 T was 2.1 Mbp with G + C content of 52.5 %, while the genome size of SYSU M7M531 was 2.3 Mbp with G + C content of 52.7 %. The ANI and 16S rRNA gene analysis results showed that the pairwise similarities between the two strains and other recognized Nitriliruptoria species were less than 64.9 % and 89.0 %, respectively. Phylogenetic analysis of the 16S rRNA gene sequences indicated that strains SYSU M7M538 T and SYSU M7M531 formed a well-separated phylogenetic branch distinct from other orders of Nitriliruptoria. Based on the data presented here, these two strains are considered to represent a novel species of a novel genus, for which the name Stomatohabitans albus gen. nov., sp. nov., with the type strain SYSU M7M538 T (=KCTC 59113 T  = GDMCC 1.4286 T ), are proposed. We also propose that these organisms represent a novel family named Stomatohabitantaceae fam. nov. of a novel order Stomatohabitantales ord. nov., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Genome mining of the Actinomycete Crossiella cryophila facilitated the discovery of a minimal terpenoid biosynthetic gene cluster of cry consisting of a class I terpene cyclase CryA and a CYP450 monooxygenase CryB. Heterologous expression of cry allowed the isolation and characterization of two new sesquiterpenoids, ent-viridiflorol (1) and cryophilain (2). Notably, cryophilain (2) possesses a 5/7/3-fused tricyclic skeleton bearing a distinctive bridgehead hydroxy group. The combined in vivo and in vitro experiments revealed that CryA, the first ent-viridiflorol terpene cyclase, catalyzes farnesyl diphosphate to form the 5/7/3 sesquiterpene core scaffold and P450 CryB serves as a tailoring enzyme responsible for installing a hydroxy group at the bridgehead carbon., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The effects of oxyanions selenite (SeO 3 2- ) in soils are of high concern in ecotoxicology and microbiology as they can react with mineral particles and microorganisms. This study investigated the evolution of the actinomycete Kitasatospora sp. SeTe27 in response to selenite. To this aim, we used the Adaptive Laboratory Evolution (ALE) technique, an experimental approach that mimics natural evolution and enhances microbial fitness for specific growth conditions. The original strain (wild type; WT) isolated from uncontaminated soil gave us a unique model system as it has never encountered the oxidative damage generated by the prooxidant nature of selenite. The WT strain exhibited a good basal level of selenite tolerance, although its growth and oxyanion removal capacity were limited compared to other environmental isolates. Based on these premises, the WT and the ALE strains, the latter isolated at the end of the laboratory evolution procedure, were compared. While both bacterial strains had similar fatty acid profiles, only WT cells exhibited hyphae aggregation and extensively produced membrane-like vesicles when grown in the presence of selenite (challenged conditions). Conversely, ALE selenite-grown cells showed morphological adaptation responses similar to the WT strain under unchallenged conditions, demonstrating the ALE strain improved resilience against selenite toxicity. Whole-genome sequencing revealed specific missense mutations in genes associated with anion transport and primary and secondary metabolisms in the ALE variant. These results were interpreted to show that some energy-demanding processes are attenuated in the ALE strain, prioritizing selenite bioprocessing to guarantee cell survival in the presence of selenite. The present study indicates some crucial points for adapting Kitasatospora sp. SeTe27 to selenite oxidative stress to best deal with selenium pollution. Moreover, the importance of exploring non-conventional bacterial genera, like Kitasatospora, for biotechnological applications is emphasized., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Actinobacteria are abundant in soil and other environmental ecosystems and are also an important part of the human microbiota. Hence, they can also be detected in indoor environments and on building materials, where actinobacterial proliferation on damp materials can indicate moisture damage. The aim of this study was to evaluate the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the identification of 28 environmental strains of Actinobacteria isolated from building materials and indoor and outdoor air samples, mainly collected in the context of moisture damage investigations in buildings in Finland. The 16S rRNA gene sequencing and chemotaxonomic analyses were performed, and results were compared with the MALDI-TOF MS Biotyper identification. Using 16S rRNA gene sequencing, all isolates were identified on the species or genus level and were representatives of Streptomyces, Nocardia, and Pseudonocardia genera. Based on MALDI-TOF MS analysis, initially, 11 isolates were identified as Streptomyces spp. and 1 as Nocardia carnea with a high identification score. After an upgrade in the MALDI-TOF MS in-house database and re-evaluation of mass spectra, 13 additional isolates were identified as Nocardia , Pseudonocardia, and Streptomyces . MALDI-TOF MS has the potential in environmental strain identification; however, the standard database needs to be considerably enriched by environmental Actinobacteria representatives., Importance: The manuscript addresses the challenges in identifying environmental bacteria using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) Biotyper-based protein profiling. The matter of the studies-actinobacterial strains-has been isolated mostly from building materials that originated from a confirmed moisture-damaged situation. Polyphasic taxonomy, 16S RNA gene sequencing, and MALDI-TOF mass spectrometry were applied for identification purposes. In this experimental paper, a few important facts are highlighted. First, Actinobacteria are abundant in the natural as well as built environment, and their identification on the species and genus levels is difficult and time-consuming. Second, MALDI-TOF MS is an effective tool for identifying bacterial environmental strains, and in parallel, continuous enrichment of the proteomics mass spectral databases is necessary for proper identification. Third, the chemical approach aids in the taxonomical inquiry of Actinobacteria environmental strains., Competing Interests: The authors declare no conflict of interest.

During the life activities of microorganisms, a variety of secondary metabolites are produced, including antimicrobials and antitumor drugs, which are widely used in clinical practice. In addition to exploring new antibiotics, this makes it one of the research priorities of Actinomycetes to effectively increase the yield of antibiotics in production strains by various means. Most antibiotic-producing strains have a variety of functional regulatory factors that regulate their growth, development, and secondary metabolite biosynthesis processes. Through the study of precursor substances in antibiotic biosynthesis, researchers have revealed the precursor biosynthesis process and the mechanism by which precursor synthesis regulators affect the biosynthesis of secondary metabolites, which can be used to obtain engineered strains with high antibiotic production. This paper summarizes the supply of antibiotic biosynthesis precursors and the progress of research on the role of regulators in the process of precursors in biosynthesis. This lays the foundation for the establishment of effective breeding methods to improve antibiotic yields through the manipulation of precursor synthesis genes and related regulators.

Objectives: This systematic review was to evaluate the change of oral microbiome based on next‐generation sequencing (NGS)‐metagenomic analysis following periodontal interventions among systematically healthy subjects. Materials and Methods: A structured search strategy consisting of "metagenomics" and "oral diseases" was applied to PubMed, EMBASE, and Web of Science to identify effective papers. The included studies were original studies published in English, using metagenomic approach to analyze the effectiveness of periodontal intervention on oral microbiome among systematically healthy human subjects with periodontitis. Results: A total of 12 papers were included in this review. Due to the heterogeneity of selected study, quantitative analysis was not performed. The findings as to how alpha diversity changed after interventions were not consistent across studies. Six studies illustrated clear separation of microbial composition between dental plaque samples collected before and after intervention using principal coordinates/component analysis. The most commonly detected genera before intervention were Porphyromonas, Treponema, Tannerella, and Prevotella, while Streptococcus and Actinomyces usually increased and became the dominant genera after intervention. Correlation network analysis revealed that after intervention, the topology of network was different compared to the corresponding pre‐interventional samples. Conclusion: Existing evidence of metagenomic studies depicts a complex change in oral microbiome after periodontal intervention. [ABSTRACT FROM AUTHOR]