Alkaliphilic Bacterium Research Articles

Distribution of Microorganisms and Antibiotic Resistance Genes in Production Wastewater During Pumped Storage Power Station Construction

The construction period of pumped storage power stations (PSPS) generates amounts of production wastewater, which may contain pathogenic bacteria and antibiotic resistance genes (ARGs) in these bacteria, potentially posing environmental and health risks. This study used the metagenome approach to analyze the distribution of microorganisms, ARGs and their correlation with water quality indicators in wastewater collected from two typical PSPSs. Coagulation system wastewater exhibits strong alkalinity (11.88), and aggregate system wastewater has high suspended solids (SS, 8 × 104 mg/L), resulting in lower richness and diversity of bacterial communities. Serpentinimonas, a kind of alkaliphilic bacteria, had the highest relative abundance (48.58–99.7%). The ARG subtypes obtained conferred wastewater resistance to tetracycline, macrolide, fluoroquinolone and so on, but wastewater treatment has limited removal effect on ARGs. The results indicate that resistant bacteria and resistance genes can still be present and distributed under highly alkaline conditions, and the removal efficiency of ARGs by wastewater treatment in PSPS is limited. Attention should be given to the environmental and health risks posed by production wastewater, thereby providing a theoretical basis for the sustainable development of the PSPS industry.

Industrial and Pharmaceutical Applications of Microbial Diversity of Hypersaline Ecology from Lonar Soda Crater.

The unidentified geochemical and physiochemical characteristics of Soda Lakes across the globe make it a novel reservoir and bring attention to scientific civic for its conceivable industrial and pharmaceutical applications. In India, in the Maharashtra state, Lonar Lake is a naturally created Soda Lake by a meteorite impact. Phylogenetic data from this lake explored a diverse array of microorganisms like haloalkaliphilic bacteria and Archaea. Previously reported studies postulated the major microbial communities present in this lake ecosystem are Proteobacteria, Actinobacteria, Firmicutes, and Cyanobacteria. Furthermore, it also contains Bacteroidetes, Nitrospirae, and Verrucomicrobia. This lake is also rich in phytoplankton, with the predominant presence of the Spirulina plantensis. Unique microbial strains from Lonar Lake ecosystems have fascinated consideration as a source of biological molecules with medicinal, industrial, and biotechnological potential. Recent literature revealed the isolation of antibioticproducing bacteria and alkaline proteases-producing alkaliphilic bacterium, as well as novel species of rare methylotrophs, other bacterial strains involved in producing vital enzymes, and unique actinomycetes are also reported. It indicates that the novel bacterial assemblage not reached hitherto may exist in this modified and unique ecology. This comprehensive review provides information about microbial diversity and its industrial and pharmaceutical interests that exist in Lonar Lake, which could be the future source of bioactive enzymes, biosurfactants, and biofuel and also useful in bioremediation. Furthermore, the novel species of microorganisms isolated from Lonar Lake have applications in the biosynthesis of medicines like antibiotics, antivirals, antifungals, anti-inflammatory agents, and precursors for synthesising valuable products. Data consolidated in the present review will cater to the needs of emerging industrial sectors for their commercial and therapeutic applications.

A protet-based model that can account for energy coupling in oxidative and photosynthetic phosphorylation

Two-stage (e.g. light-dark) phosphorylation experiments showed that there is a stored ‘high-energy’ intermediate linking electron transport and phosphorylation. Large, artificial electrochemical proton gradients (protonmotive forces or pmfs) can also drive phosphorylation, a fact seen as strongly supportive of the chemiosmotic coupling hypothesis that a pmf is the ‘high-energy’ intermediate. However, in such experiments there is an experimental threshold (pmf >170 mV, equivalent to ΔpH ∼2.8) below which no phosphorylation is in fact observed, and 220 mV are required to recreate in vivo rates. This leads to the correct question, which is then whether those values of the pmf generated by electron transport are large enough. Even the lower ones as required for any phosphorylation (leave alone those required to explain in vivo rates) are below the threshold [1, 2], whether measured directly with microelectrodes or via the use of membrane-permeant ions and/or acids/bases (which are always transporter substrates [3], so all such measurements are in fact artefactual). The single case that seemed large enough (220 mV) is now admitted to be a diffusion potential artefact [4]. Many other observables (inadequate bulk H+ in ‘O2-pulse’-type experiments, alkaliphilic bacteria, dual-inhibitor titrations, uncoupler-binding proteins, etc.) are consistent with the view that values of the pmf, and especially of Δψ, are actually very low. A protet-based charge separation model [2], a protonic version analogous to how energy may be stored in devices called electrets, provides a high-energy intermediate that can explain the entire literature, including the very striking demonstration [5] that close proximity is required between electron transport and ATP synthase complexes for energy coupling between them to allow phosphorylation to occur. A chief purpose of this article is thus to summarise the extensive and self-consistent literature, much of which is of some antiquity and rarely considered by modern researchers, despite its clear message of the inadequacy of chemiosmotic coupling to explain these phenomena.

Potential of halophiles and alkaliphiles in bioremediation of azo dyes-laden textile wastewater: a review.

Azo dye-laden textile wastewater must be treated before release due to various health and environmental concerns. Bioremediation of textile wastewater, however, is a challenge owing to its alkaline and saline nature as mesophilic microbes, in general, are either not able to thrive or show less efficiency under such hostile environment. Thus, pre-treatment for neutralization or salinity removal becomes a prerequisite before applying microbes for treatment, causing extra economical and technical burden. Extremophilic bacteria can be the promising bioremediating tool because of their inherent ability to survive and show toxicants removal capability under such extreme conditions without need of pre-treatment. Among extremophiles, halophilic and alkaliphilic bacteria which are naturally adapted to high salt and pH are of special interest for the decolorization of saline-alkaline-rich textile wastewater. The current review article is an attempt to provide an overview of the bioremediation of azo dyes and azo dye-laden textile wastewater using these two classes of extremophilic bacteria. The harmful effects of azo dyes on human health and environment have been discussed herein. Halo-alkaliphilic bacteria circumvent the extreme conditions by various adaptations, e.g., production of certain enzymes, adjustment at the protein level, pH homeostasis, and other structural adaptations that have been highlighted in this review. The unique properties of alkaliphiles and halophiles, to not only sustain but also harboring high dye removal competence at high pH and salt concentration, make them a good candidate for designing future bioremediation strategies for the management of alkaline, salt, and azo dye-laden industrial wastewaters.

Exogenous putrescine plays a switch-like influence on the pH stress adaptability of biofilm-based activated sludge.

Microbial community adaptability to pH stress plays a crucial role in biofilm formation. This study aims to investigate the regulatory mechanisms of exogenous putrescine on pH stress, as well as enhance understanding and application for the technical measures and molecular mechanisms of biofilm regulation. Findings demonstrated that exogenous putrescine acted as a switch-like distributor affecting microorganism pH stress, thus promoting biofilm formation under acid conditions while inhibiting it under alkaline conditions. As pH decreases, the protonation degree of putrescine increases, making putrescine more readily adsorbed. Protonated exogenous putrescine could increase cell membrane permeability, facilitating its entry into the cell. Subsequently, putrescine consumed intracellular H+ by enhancing the glutamate-based acid resistance strategy and the γ-aminobutyric acid metabolic pathway to reduce acid stress on cells. Furthermore, putrescine stimulated ATPase expression, allowing for better utilization of energy in H+ transmembrane transport and enhancing oxidative phosphorylation activity. However, putrescine protonation was limited under alkaline conditions, and the intracellular H+ consumption further exacerbated alkali stress and inhibits cellular metabolic activity. Exogenous putrescine promoted the proportion of fungi and acidophilic bacteria under acidic stress and alkaliphilic bacteria under alkali stress while having a limited impact on fungi in alkaline biofilms. Increasing Bdellovibrio under alkali conditions with putrescine further aggravated the biofilm decomposition. This research shed light on the unclear relationship between exogenous putrescine, environmental pH, and pH stress adaptability of biofilm. By judiciously employing putrescine, biofilm formation could be controlled to meet the needs of engineering applications with different characteristics.IMPORTANCEThe objective of this study is to unravel the regulatory mechanism by which exogenous putrescine influences biofilm pH stress adaptability and understand the role of environmental pH in this intricate process. Our findings revealed that exogenous putrescine functioned as a switch-like distributor affecting the pH stress adaptability of biofilm-based activated sludge, which promoted energy utilization for growth and reproduction processes under acidic conditions while limiting biofilm development to conserve energy under alkaline conditions. This study not only clarified the previously ambiguous relationship between exogenous putrescine, environmental pH, and biofilm pH stress adaptability but also offered fresh insights into enhancing biofilm stability within extreme environments. Through the modulation of energy utilization, exerting control over biofilm growth and achieving more effective engineering goals could be possible.

Roseinatronobacter alkalisoli sp. nov., an alkaliphilic bacterium isolated from soda soil, and genome-based reclassification of the genera Rhodobaca and Roseinatronobacter.

The genera Rhodobaca and Roseinatronobacter are phylogenetically related genera within the family Paracoccaceae. Species of these genera were described using 16S rRNA gene-based phylogeny and phenotypic characteristics. However, the 16S rRNA gene identity and phylogeny reveal the controversy of the taxonomic status of these two genera. In this work, we examined the taxonomic positions of members of both genera using 16S rRNA gene phylogeny, phylogenomic analysis and further validated using overall genome-related indexes, including digital DNA-DNA hybridization, average nucleotide identity, average amino acid identity and percentage of conserved proteins. Based on phylogenetic and phylogenomic results, the current four species of the two genera clustered tightly into one clade with high bootstrap values, suggesting that the genus Rhodobaca should be merged with Roseinatronobacter. In addition, a novel species isolated from a soda soil sample collected from Anda City, PR China, and designated as HJB301T was also described. Phenotypic, chemotaxonomic, genomic and phylogenetic properties suggested that strain HJB301T (=CCTCC AB 2021113T=KCTC 82977T) represents a novel species of the genus Roseinatronobacter, for which the name Roseinatronobacter alkalisoli sp. nov. is proposed.

Heteroxylan hydrolysis by a recombinant cellulase-free GH10 xylanase from the alkaliphilic bacterium Halalkalibacterium halodurans C-125.

The search for affordable enzymes with exceptional characteristics is fundamental to overcoming industrial and environmental constraints. In this study, a recombinant GH10 xylanase (Xyn10-HB) from the extremelyalkaliphilic bacterium Halalkalibacteriumhalodurans C-125 cultivated at pH 10 was cloned and expressed in E. coli BL21(DE3). Removal of the signal peptide improved the expression, and an overall activity of 8 U/mL was obtained in the cell-free supernatant. The molecular weight of purified Xyn10-HB was estimated to be 42.6 kDa by SDS-PAGE. The enzyme was active across a wide pH range (5-10) with optimal activity recorded at pH 8.5 and 60°C. It also presented good stability with a half-life of 3h under these conditions. Substrate specificity studies showed that Xyn10-HB is a cellulase-free enzyme that conventionally hydrolyse birchwood and oat spelts xylans (Apparent Km of 0.46mg/mL and 0.54mg/mL, respectively). HPLC analysis showed that both xylans hydrolysis produced xylooligosaccharides (XOS) with a degree of polymerization (DP) ranging from 2 to 9. The conversion yield was 77% after 24h with xylobiose and xylotriose as the main end-reaction products. When assayed on alkali-extracted wheat straw heteroxylan, the Xyn10-HB produced active XOS with antioxidant activity determined by the DPPH radical scavenging method (IC50 of 0.54mg/mL after 4h). Owing to its various characteristics, Xyn10-HB xylanase is a promising candidate for multiple biotechnological applications.

Expression and biochemical characterization of a novel thermostable alkaline β-1,3–1,4-glucanase (lichenase) from an alkaliphilic Bacillus lehensis G1

New thermostable β-1,3–1,4-glucanase (lichenase) designated as Blg29 was expressed and purified from a locally isolated alkaliphilic bacteria Bacillus lehensis G1. The genome sequence of B. lehensis predicted an open reading frame of Blg29 with a deduced of 249 amino acids and a molecular weight of 28.99 kDa. The gene encoding for Blg29 was successfully amplified via PCR and subsequently expressed as a recombinant protein using the E. coli expression system. Recombinant Blg29 was produced as a soluble form and further purified via immobilized metal ion affinity chromatography (IMAC). Based on biochemical characterization, recombinant Blg29 showed optimal activity at pH9 and temperature 60 °C respectively. This enzyme was stable for more than 2 h, incubated at 50 °C, and could withstand ∼50 % of its activity at 70 °C for an hour and a half. No significant effect on Blg29 was observed when incubated with metal ions except for a small increase with ion Ca2+. Blg29 showed high substrate activity towards lichenan where Vm, Km, Kcat, and kcat/Km values were 2040.82 μmolmin‾1mg‾1, 4.69 mg/mL, and 986.39 s‾1 and 210.32 mLs‾1mg‾1 respectively. The high thermostability and activity make this enzyme useable for a broad prospect in industry applications.

Comparative analysis of reconstructed ancestral proteins with their extant counterparts suggests primitive life had an alkaline habitat

To understand the origin and early evolution of life it is crucial to establish characteristics of the primordial environment that facilitated the emergence and evolution of life. One important environmental factor is the pH of the primordial environment. Here, we assessed the pH-dependent thermal stabilities of previously reconstructed ancestral nucleoside diphosphate kinases and ribosomal protein uS8s. The selected proteins were likely to be present in ancient organisms such as the last common ancestor of bacteria and that of archaea. We also assessed the thermal stability of homologous proteins from extant acidophilic, neutralophilic, and alkaliphilic microorganisms as a function of pH. Our results indicate that the reconstructed ancestral proteins are more akin to those of extant alkaliphilic bacteria, which display greater stability under alkaline conditions. These findings suggest that the common ancestors of bacterial and archaeal species thrived in an alkaline environment. Moreover, we demonstrate the reconstruction method employed in this study is a valuable technique for generating alkali-tolerant proteins that can be used in a variety of biotechnological and environmental applications.

Genomic Insights into Syntrophic Lifestyle of 'Candidatus Contubernalis alkaliaceticus' Based on the Reversed Wood-Ljungdahl Pathway and Mechanism of Direct Electron Transfer.

The anaerobic oxidation of fatty acids and alcohols occurs near the thermodynamic limit of life. This process is driven by syntrophic bacteria that oxidize fatty acids and/or alcohols, their syntrophic partners that consume the products of this oxidation, and the pathways for interspecies electron exchange via these products or direct interspecies electron transfer (DIET). Due to the interdependence of syntrophic microorganisms on each other's metabolic activity, their isolation in pure cultures is almost impossible. Thus, little is known about their physiology, and the only available way to fill in the knowledge gap on these organisms is genomic and metabolic analysis of syntrophic cultures. Here we report the results of genome sequencing and analysis of an obligately syntrophic alkaliphilic bacterium 'Candidatus Contubernalis alkaliaceticus'. The genomic data suggest that acetate oxidation is carried out by the Wood-Ljungdahl pathway, while a bimodular respiratory system involving an Rnf complex and a Na+-dependent ATP synthase is used for energy conservation. The predicted genomic ability of 'Ca. C. alkaliaceticus' to outperform interspecies electron transfer both indirectly, via H2 or formate, and directly, via pili-like appendages of its syntrophic partner or conductive mineral particles, was experimentally demonstrated. This is the first indication of DIET in the class Dethiobacteria.

The genus Anoxybacillus: an emerging and versatile source of valuable biotechnological products.

Thermophilic and alkaliphilic microorganisms are unique organisms that possess remarkable survival strategies, enabling them to thrive on a diverse range of substrates. Anoxybacillus, a genus of thermophilic and alkaliphilic bacteria, encompasses 24 species and 2 subspecies. In recent years, extensive research has unveiled the diverse array of thermostable enzymes within this relatively new genus, holding significant potential for industrial and environmental applications. The biomass of Anoxybacillus has demonstrated promising results in bioremediation techniques, while the recently discovered metabolites have exhibited potential in medicinal experiments. This review aims to provide an overview of the key experimental findings related to the biotechnological applications utilizing bacteria from the Anoxybacillus genus.

Cyclodextrin glucanotransferase: fundamentals and biotechnological implications.

Cyclodextrin glucanotransferase (CGTase) is an extracellular enzyme of the GH13 α-amylase family that catalyzes a unique intramolecular reaction known as cyclization to transform α-1, 4-glucans and similar starches into cyclodextrins. They also catalyze intermolecular transglycosylation reactions namely coupling, disproportionation, and some hydrolyzing effects on starch. The monomeric structures of the CGTase exhibit five domains (A, B, C, D, and E domains) with different molecular weights and amino acid sequences depending on the source. Among bacteria, Bacillus genus covers approximately 90% of the CGTase producers, while other genera like Klebsiella, Paenibacillus, and Thermoanaerobacter also shown decent contributions in recent studies. CGTase production is highly supported by alkaliphilic bacteria under submerged fermentation rather than solid-state fermentation. The bacterial sources, biochemical properties, production conditions, and structure of CGTases are compiled in this review. Cyclodextrins have the unique property of making inclusion complexes with various compounds, hence widely used in the food, pharmaceutical, cosmetics, laundry, and chemical sectors. This review presents a comprehensive view of CGTase produced by Bacillus spp., and other bacterial genera like Klebsiella, Paenibacillus, and Microbacterium. It also gives insight of the properties and recent biotechnological applications of cyclodextrins. KEY POINTS: • Transglycosylation reactions catalyzed by CGTase and their structural properties. • Comparative data of CGTase production by various genera and Bacillus spp. • Structures, properties, and applications of different cyclodextrins.

A novel neutrophilic bacteria based process for selective LiFePO4 cathode recycling

Battery recycling has attracted attention as a solution to maintain the chain of demand and supply. First time, a selective biological process assisted by a neutrophilic microorganism for lithium recovery from Li-FP battery cathodes is exhibited. Thiobacillus thioparus, a biosafety level-1 bacterium is used to selectively leach lithium at near-neutral pH. The study showed the leaching was efficient, selective, and environmentally friendly as it could recover 65–98% lithium, with negligible iron, under different experimental conditions supported solely by bacterial activity. Thereafter, lithium was recovered as Li3PO4 from the leach solution, which was subsequently reuse as a culture medium to grow an alkaliphilic bacteria for other applications. The liquid and solid residues were characterized by FAAS, ICP-MS, XRD, FESEM, XPS, UV–visible, and FTIR techniques. The developed process could suggest a sustainable alternative for reclaiming selective recovery of critical metal, which needs to meet the demand for continuous consumption in battery production.

Cover: Journal of Basic Microbiology. 3‐4/2023

Journal of Basic MicrobiologyVolume 63, Issue 3-4 p. 235-235 COVER PICTUREFree Access Cover: Journal of Basic Microbiology. 3-4/2023 First published: 04 March 2023 https://doi.org/10.1002/jobm.202370005AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Graphical Abstract Cover illustration: Secretion of industrially important amylase on starch agar medium by the salt-tolerant alkaliphilic bacteria isolated from the rhizosphere of medicinal plant Eucalyptus polybractea L., collected from Kharaghoda, near desert Kutch, Gujarat, India. (Photo: Nishtha R. Vaghela, Department of Biosciences, Saurashtra University, Rajkot, Gujarat, India) Volume63, Issue3-4Special Issue: Microbes in action: Ecology to economyMarch-April 2023Pages 235-235 RelatedInformation

Genomic insights of an alkaliphilic bacterium Halalkalibacter alkaliphilus sp. nov. isolated from an Indian Soda Lake.

An alkaliphilic, Gram-stain-positive, non-motile, rod-shaped, and spore forming bacterial strain (MEB205T) was isolated from sediment sample collected from Lonar lake, India. The strain grew optimally at pH 10, NaCl concentration of 3.0% at 37°C. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain MEB205T belonged to the genus Halalkalibacter in the family Bacillaceae and shared the highest sequence similarity with H. okhensis Kh10-101T (98.9%) followed by H. wakoensis N-1T (98.7%). The assembled genome of strain MEB205T has a total length of 4.8Mb with a G + C content of 37.8%. The dDDH and OrthoANI values between strain MEB205T and H. okhensis Kh10-101T were 29.1% and 84.3%, respectively. Furthermore, the genome analysis revealed the presence of antiporter genes (nhaA and nhaD) and L-ectoine biosynthesis gene required for survival of the strain MEB205T in alkaline-saline habitat. The major fatty acid was C15:0 anteiso, C16:0 and C15:0 iso (> 10.0%). Diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine were the major polar lipids. meso-diaminopimelic acid was diagnostic diamino acid for cell wall peptidoglycan. Based on the polyphasic taxonomic studies, strain MEB205T represent a novel species of the genus Halalkalibacter for which the name Halalkalibacter alkaliphilus sp. nov. (Type strain MEB205T = MCC 3863T = JCM 34004T = NCIMB 15406T) is proposed.

Bacillus Swezeyi B2 Strain: A Novel Alkaliphilic Bacterium Producer of Alkaline-, Thermal, Oxidant-, and Surfactant-Stable Protease, Extremely Efficient in Detergency.

Proteolytic enzymes that are currently used to meet industrial demand are usually derived from Bacillus species. They find multiple technical applications, particularly they have been increasingly used as a key bio-additive in detergents. In this study, a novel alkalophilic bacterium was isolated from contaminated soil, exhibiting 1400U/ml proteolytic activity, and identified as Bacillus swezeyi B2. The crude enzyme likely contained a single extracellular protease. This enzyme revealed optimum activity at pH 10 and 70°C and was highly alkaline thermostable (7-12.5) and up to 70°C. The protease activity was completely inhibited by Phenylmethylsulfonyl fluoride (PMSF) suggesting that it belongs to the serine protease group. It was highly stable in the presence of the strong anionic surfactant (SDS) and oxidizing agents (H2O2). The supernatant was lyophilized and showed high storage stability retaining 100% of its original activity after one year of conservation at 4°C. The lyophilized product was evaluated for its detergent efficacy, it revealed excellent compatibility with various laundry detergents keeping its full original activity after incubation for 1h with seven solid and liquid commercial detergents and it effectively removed chocolate stains at low washing temperature (40°C) and low supplementation level (125U/ml). The features of this single alkaline and thermotolerant protease, stable toward surfactants, oxidizing agents, and commercial detergents with stain removal efficacy support its ideal choice for supplementation in detergent formulations.

Nitrogen and boron co-doped lignin biochar for enhancing calcium peroxide activation toward organic micropollutants decontamination in waste activated sludge and related microbial structure dynamics

This study synthesized dual heteroatom nitrogen and boron-co-doped lignin-based biochar (NB-LGBC) for calcium peroxide (CP) activation to enhance the removal of organic micropollutants (OMPs), namely, 4-nonylphenol (4-NP) from waste activated sludge (WAS). NB-LGBC/CP enhanced 4-NP degradation by arriving at 83 % removal in 12 h. The NB-LGBC/CP system degraded 4-NP via a synergistic interaction (HO•, O2•− radicals, and singlet oxygen) and electron transfer due to the N–B–C bonding configurations. Results of fluorescence excitation-emission matrix (FEEM) analysis revealed significantly increase in biodegradable organics from treated WAS mixture. NB-LGBC/CP treatment enriched alkaliphilic bacterium associated with the predominance of the genus Desulfonatronum within the phylum Proteobacteria in the WAS, which improved the biological treatment capacity of 4-NP. Thus, NB-LGBC in HR-CAOP will be a novel approach for WAS decontamination.

Ion transfer mechanisms in Mrp-type antiporters from high resolution cryoEM and molecular dynamics simulations

Multiple resistance and pH adaptation (Mrp) cation/proton antiporters are essential for growth of a variety of halophilic and alkaliphilic bacteria under stress conditions. Mrp-type antiporters are closely related to the membrane domain of respiratory complex I. We determined the structure of the Mrp antiporter from Bacillus pseudofirmus by electron cryo-microscopy at 2.2 Å resolution. The structure resolves more than 99% of the sidechains of the seven membrane subunits MrpA to MrpG plus 360 water molecules, including ~70 in putative ion translocation pathways. Molecular dynamics simulations based on the high-resolution structure revealed details of the antiport mechanism. We find that switching the position of a histidine residue between three hydrated pathways in the MrpA subunit is critical for proton transfer that drives gated trans-membrane sodium translocation. Several lines of evidence indicate that the same histidine-switch mechanism operates in respiratory complex I.

Bioconversion of lignocellulosic wastes to zwitterionic biosurfactants by an alkaliphilic bacterium: Process development and product characterization

The bioconversion of lignocellulosic wastes to value-added biosurfactants was investigated using a single flask to carry out both substrate pretreatment and biosurfactant production. Lignocellulosic wastes were added to an alkaline medium and autoclaved, which led to a combined alkaline and hydrothermal pretreatment. For biosurfactant production, alkaliphilic Brevibacterium casei NK8 was added to the medium without pH neutralization. Among several lignocellulosic wastes, the media with corn husk, coconut oil cake, and defatted rice bran gave the highest biosurfactant concentrations (1.14–1.32 g L−1). These wastes provided a relatively high concentration of soluble substrates in the pretreated liquid, while solid residues swelled. The bacterium could use the pretreated liquid and solid residues for growth and biosurfactant production. The produced biosurfactants in cell-free broth could be applied directly or after lignin removal. The residual lignin decreased biosurfactant performance in terms of surface tension, contact angle, and oil displacement; conversely, lignin enhanced the biosurfactant emulsification capacity. Cell-free broths with and without lignin were appropriate for different tasks. Biosurfactants synthesized from different lignocellulosic wastes were similar and were identified as a mixture of zwitterionic lipopeptides and phospholipids. The surface tension of the crude biosurfactant solution was extremely low, ranging between 25.9 and 27.6 mN m−1 over a wide range of pH values (1–11), salinities (2–10% NaCl), and temperatures (4–121 °C). In addition, biosurfactants as crude solution and cell-free broth with lignin showed lower phytotoxicity than the commercial surfactant Tween 80. These findings suggest a convenient method for manufacturing biosurfactants from lignocellulosic wastes.

Isolation, purification and biochemical characterization of a thermophilic alkaline protease from hot water spring bacteria

Search for a new alkaline and thermostable protease is crucial to explore their industrial applications. Therefore, a thermophilic, alkaliphilic bacteria producing serine alkaline protease was isolated from a hot water spring. Isolated bacteria were identified as B. licheniformis strain 9AkScKu3 as per the results revealed from biochemical studies and 16s rRNA gene sequencing analysis. The purification fold of the enzyme was 23.91 with yield of 18.54%. The purified protease was optimally active at pH 9 and a temperature of 50°C. The enzyme exhibited thermal stability up to 70ºC. Metal ions Ca2+, Mg2+ and Cu2+ enhanced the activity of enzyme. The enzyme exhibited compatibility with 3% surfactants showing more than 100% stability with SDS, tween 20 and triton-x100. Enzyme retained 100% activity with 5% (v/v) H2O2 and organic solvents like toluene and n-hexane at 10% (v/v) concentration. Thus isolated thermophilic alkaline protease is a promising enzyme applicable in detergent formulation.