Bacillus cereus G2 alleviate salt stress in Glycyrrhiza uralensis Fisch. by balancing the downstream branches of phenylpropanoids and activating flavonoid biosynthesis.

The salinity environment is one of the biggest threats to Glycyrrhiza uralensis Fisch. (G. uralensis) growth, resulting from the oxidative stress caused by excess reactive oxygen species (ROS). Flavonoids are the main pharmacodynamic composition and help maintain ROS homeostasis and mitigate oxidative damage in G. uralensis in the salinity environment. To investigate whether endophytic Bacillus cereus G2 can improve the salt-tolerance of G. uralensis through controlling flavonoid biosynthesis, the transcriptomic and physiological analysis of G. uralensis treated by G2 in the saline environment was conducted, focused on flavonoid biosynthesis-related pathways. Results uncovered that salinity inhibited flavonoids synthesis by decreasing the activities of phenylalanine ammonialyase (PAL) and 4-coumarate-CoA ligase (4CL) (42% and 39%, respectively) due to down-regulated gene Glyur000910s00020578 at substrate level, and then decreasing the activities of chalcone isomerase (CHI) and chalcone synthase (CHS) activities (50% and 42%, respectively) due to down-regulated genes Glyur006062s00044203 and Glyur000051s00003431, further decreasing isoliquiritigenin content by 53%. However, salt stress increased liquiritin content by 43%, which might be a protective mechanism of salt-treated G. uralensis seedlings. Interestingly, G2 enhanced PAL activity by 27% whereas reduced trans-cinnamate 4-monooxygenase (C4H) activity by 43% which could inhibit lignin biosynthesis but promote flavonoid biosynthesis of salt-treated G. uralensis at the substrate level. G2 decreased shikimate O-hydroxycinnamoyltransferase (HCT) activity by 35%, increased CHS activity by 54% through up-regulating the gene Glyur000051s00003431 encoding CHS, and increased CHI activity by 72%, thereby decreasing lignin (34%) and liquiritin (24%) content, but increasing isoliquiritigenin content (35%), which could mitigate oxidative damage and changed salt-tolerance mechanism of G. uralensis.

Association of Laccase from Bacillus cereus O2-B and Pseudomonas aeruginosa O1-P with the bio-degradation of polymers: an in vitro to in silico approach.

Plastic accumulation has become a serious environmental threat. Mitigation of plastic is important to save the ecosystem of our planet. With current research being focused on microbial degradation of plastics, microbes with the potential to degrade polyethylene were isolated in this study. In vitro studies were performed to define the correlation between the degrading capability of the isolates and laccase, a common oxidase enzyme. Instrumental analyses were used to evaluate morphological and chemical modifications in polyethylene, which demonstrated a steady onset of the degradation process in case of both isolates, Pseudomonas aeruginosa O1-P and Bacillus cereus O2-B. To understand the efficiency of laccase in degrading other common polymers, in silico approach was employed, for which 3D structures of laccase in both the isolates were constructed via homology modeling and molecular docking was performed, revealing that the enzyme laccase can be exploited to degrade a wide range of polymers.

Sustained degradation of phenol under extreme conditions by polyurethane-based Bacillus sp. ZWB3.

Phenol is a serious pollutant to the environment, therefore, it is urgent to find a rapid and effective method for its removal. In this study, Bacillus cereus ZWB3 immobilized on a polyurethane (PUF) carrier was studied. The PUF-ZWB3 required only 20 h for the degradation of 1,500 mg L-1 of phenol, shortened by 8 h than the free bacteria. In addition, the PUF-ZWB3 could increase the degradation concentration of phenol from 1,500 to 2,000 mg L-1, and the complete degradation of 2,000 mg L-1 phenol only used 44 h. In addition, the PUF-ZWB3 showed much higher removal of phenol than the free bacteria at different pH values, salt concentrations, and heavy metal ions. Particularly, the PUF-ZWB3 could still completely remove phenol in a strongly alkaline environment, such as pH 10 and 11. In addition, the removal efficiency of phenol by PUF-ZWB3 was still 100% after 10 cycles. This study showed that the PUF immobilization system had great potential in the field of remediation of organic pollution.

Production of surfactant-stable keratinase from Bacillus cereus YQ15 and its application as detergent additive.

BACKGROUND: With the growing concern for the environment, there are trends that bio-utilization of keratinous waste by keratinases could ease the heavy burden of keratinous waste from the poultry processing and leather industry. Especially surfactant-stable keratinases are beneficial for the detergent industry. Therefore, the production of keratinase by Bacillus cereus YQ15 was improved; the characterization and use of keratinase in detergent were also studied. RESULTS: A novel alkaline keratinase-producing bacterium YQ15 was isolated from feather keratin-rich soil and was identified as Bacillus cereus. Based on the improvement of medium components and culture conditions, the maximum keratinase activity (925 U/mL) was obtained after 36 h of cultivation under conditions of 35 °C and 160 rpm. Moreover, it was observed that the optimal reacting temperature and pH of the keratinase are 60 °C and 10.0, respectively; the activity was severely inhibited by PMSF and EDTA. On the contrary, the keratinase showed remarkable stability in the existence of the various surfactants, including SDS, Tween 20, Tween 60, Tween 80, and Triton X-100. Especially, 5% of Tween 20 and Tween 60 increased the activity by 100% and 60%, respectively. Furtherly, the keratinase revealed high efficiency in removing blood stains. CONCLUSION: The excellent compatibility with commercial detergents and the high washing efficiency of removing blood stains suggested its suitability for potential application as a bio-detergent additive.

De novo genome assembly and comparative annotation reveals metabolic versatility in cellulolytic bacteria from cropland and forest soils.

Cellulose, the most abundant polysaccharide in nature, is a rich source of renewable energy and sustains soil nutrients. Among the microorganisms known to degrade cellulose, bacteria are less studied compared to fungi. In the present work, we have investigated the culturable bacteria actively involved in cellulose degradation in forest and crop field soils. Based on clear zone formation and enzyme activity assay, we identified 7 bacterial strains positive for cellulose degradation. Of these, two most efficient strains (Bacillus cereus strains BHU1 and BHU2) were selected for whole genome sequencing, annotation, and information regarding GC content, number of genes, total subsystems, starch, and cellulose degradation pathways. Average nucleotide identity (ANI) showed more than 90% similarity between both the strains (BHU1 and BHU2) and with B. cereus ATCC 14579. Both the strains have genes and enzyme families like endoglucanase and ß-glucosidase as evident from whole genome sequence. Cellulase containing gene families (GH5, GH8, GH1), and many other carbohydrate-degrading enzymes, were present in both the bacterial strains. Taken together, the results suggest that the strains were efficient in cellulose degradation, and can be used for energy generation and production of value-added product.

Proteome profile changes during poly-hydroxybutyrate intracellular mobilization in gram positive Bacillus cereus tsu1.

BACKGROUND: Bacillus cereus is a bacterial species which grows efficiently on a wide range of carbon sources and accumulates biopolymer poly-hydroxybutyrate (PHB) up to 80% cell dry weight. PHB is an aliphatic polymer produced and stored intracellularly as a reservoir of carbon and energy, its mobilization is a key biological process for sporulation in Bacillus spp. Previously, B. cereus tsu1 was isolated and cultured on rapeseed cake substrate (RCS), with maximum of PHB accumulation reached within 12 h, and depleted after 48 h. Fore-spore and spore structure were observed after 24 h culture. RESULTS: Quantitative proteomic analysis of B. cereus tsu1 identified 2952 quantifiable proteins, and 244 significantly changed proteins (SCPs) in the 24 h:12 h pair of samples, and 325 SCPs in the 48 h:12 h pair of samples. Based on gene ontology classification analysis, biological processes enriched only in the 24 h:12 h SCPs include purine nucleotide metabolism, protein folding, metal ion homeostasis, response to stress, carboxylic acid catabolism, and cellular amino acid catabolism. The 48 h:12 h SCPs were enriched into processes including carbohydrate metabolism, protein metabolism, oxidative phosphorylation, and formation of translation ternary structure. A key enzyme for PHB metabolism, poly(R)-hydroxyalkanoic acid synthase (PhaC, KGT44865) accumulated significantly higher in 12 h-culture. Sporulation related proteins SigF and SpoEII were significantly higher in 24 h-samples. Enzymes for nitrate respiration and fermentation accumulated to the highest abundance level in 48 h-culture. CONCLUSIONS: Changes in proteome of B. cereus tsu1 during PHB intracellular mobilization were characterized in this study. The key enzyme PhaC for PHB synthesis increased significantly after 12 h-culture which supports the highest PHB accumulation at this time point. The protein abundance level of SpoIIE and SigF also increased, correlating with sporulation in 24 h-culture. Enzymes for nitrate respiration and fermentation were significantly induced in 48 h-culture which indicates the depletion of oxygen at this stage and carbon flow towards fermentative growth. Results from this study provide insights into proteome profile changes during PHB accumulation and reuse, which can be applied to achieve a higher PHB yield and to improve bacterial growth performance and stress resistance.

Biosynthesis and accumulation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-polyethylene glycol, a hybrid co-polymer by endophytic Bacillus cereus RCL 02.

Co-polymerization of microbial polyesters, polyhydroxyalkanoates (PHAs), with synthetic polymers has become an established and promising tool in the recent past for improving the material and biological properties of the biopolyesters. Bacillus cereus RCL 02, a leaf endophytic bacterium of the oleaginous plant Ricinus communis L., has been reported to produce a significant amount of poly(3-hydroxybutyrate) [P(3HB)] and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] under batch cultivation. The present study demonstrates the synthesis and accumulation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-polyethylene glycol [P(3HB-co-3HV)-PEG] co-polymer by the isolate RCL 02 in glucose, valeric acid, and PEG-200 supplemented mineral salts medium following dual-step cultivation. The identity of P(3HB-co-3HV)-PEG co-polymer so produced has been confirmed by X-ray diffraction (XRD) analysis, Fourier-transform infrared (FTIR), and proton nuclear magnetic resonance (1H NMR) spectroscopic studies, and the purified co-polymer was found to be composed of 3.2 mol% ethylene glycol (EG) and 8.4 mol% 3HV along with 3HB. While the thermogravimetric analysis (TGA) revealed that P(3HB-co-3HV)-PEG films degraded at 269.32 °C, differential scanning calorimetry (DSC) recorded the melting peak of the co-polymer at 163.8 °C. This study emphasized to explore the endophytic Bacillus spp. for production of P(3HB-co-3HV)-PEG co-polymers with improved material properties which may find possible application for biomedical purposes.

Production, optimization and characterization of polyhydroxybutyrate, a biodegradable plastic by Bacillus spp.

Poly-ß-hydroxybutyrate (PHB) is the intracellular lipid reserve accumulated by many bacteria. The most potent terrestrial bacterium Bacillus cereus SE-1 showed more PHB accumulating cells (22.1 and 40% after 48 and 72 h) than that of the marine Bacillus sp. CS-605 (5 and 33% after 48 and 72 h). Both the isolates harbored phbB gene and the characteristics C=O peak was observed in the extracted PHB by Fourier transformed infrared spectroscopy analysis. Maltose was found to be the most suitable carbon source for the accumulation of PHB in B. cereus SE-1. The extracted PHB sample from B. cereus SE-1 was blended with a thermoplastic starch (TS) and an increased thermoplasticity and decreased crystallinity were observed after blending in comparison to the standard PHB. The melting temperature (Tm), melting enthalpy (∆Hf), and crystallinity (Xc) of the blended PHB sample were found to be 109.4 °C, 64.58 J/g, and 44.23%, respectively.

Cost-effective-substrates for production of poly-ß-hydroxybutyrate by a newly isolated Bacillus cereus PS-10.

Poly-ß-hydroxybutyrate (PHB) may serve as one of the imperative substitutes for petroleum derived plastics because of their close functional analogy and biodegradation quality. In the present study, PHB producing ability of bacterial isolates was examined on low-cost agro industrial residues. Isolate PS-10 from domestic waste landfills, identified as Bacillus cereus PS-10 produced and accumulated appreciable amount of PHB. Bacillus cereus PS-10 was capable of using a wide variety of agro-based residues viz. maize bran, rice husk, wood waste, molasses, whey etc. as cost-effective carbon sources for PHB production. Molasses at 3% (w/v) supported maximum PHB production (9.5 gl(-1)) and was followed by glycerol (8.9 gl(-1)) at 2% (w/v). Certain carbon sources like almond shell powder and walnut shell powder are being reported for the first time for PHB production and supported reasonable PHB yield i.e. 6.6 and 4.6 gl(-1), respectively. Different cost-effective nitrogen sources like corn steep liquor, chick pea bran, soy bean meal, mustard cake etc. were used for PHB production. Highest PHB production was observed at pH 7 (9.6 gl(-1)) after 48 hrs of fermentation, although B. cereus PS-10 grew and produced PHB over pH range of 5-9. Optimum inoculum level for maximum PHB production was found to be 5% v/v (A600 0.9; approximately 10(8) cfu ml(-1)). Fourier transform infrared spectroscopy (FT-IR) analysis of the extracted PHB showed characteristic peaks (1721.95, 1632.19 and 2926.43 cm(-1)) similar to standard PHB. Melting point of PHB was found to be 185°C. Bacillus cereus PS-10 may be a sound PHB producer, especially by exploiting low cost substrates.

Enhanced biosynthesis of poly(3-hydroxybutyrate) from potato starch by Bacillus cereus strain 64-INS in a laboratory-scale fermenter.

To decrease the polyhydroxyalkanoate (PHA) production cost by supplying renewable carbon sources has been an important aspect in terms of commercializing this biodegradable polymer. The production of biodegradable poly(3-hydroxyalkanoates) (PHA) from raw potato starch by the Bacillus cereus 64-INS strain isolated from domestic sludge has been studied in a lab-scale fermenter. The bacterium was screened for the degradation of raw potato starch by a starch hydrolysis method and for PHA production by Nile blue A and Sudan black B staining. Shake-flask cultures of the bacterium with glucose [2% (w/v)] or raw potato starch [2% (w/v)] produced PHA of 64.35% and 34.68% of dry cell weight (DCW), respectively. PHA production was also carried out in a 5-L fermenter under control conditions that produced 2.78 g/L of PHA and PHA content of 60.53% after 21 hr of fermentation using potato starch as the sole carbon source. Gas chromatography-mass spectroscopy (GC-MS) analyses confirmed that the extracted PHA contained poly(3-hydroxybutyrate) (PHB) as its major constituent (>99.99%) irrespective of the carbon source used. The article describes, for what we believe to be the first time, PHB production being carried out without any enzymatic or chemical treatment of potato starch at higher levels by fermentation. More work is required to optimize the PHB yield with respect to starch feeding strategies.

Microbial degradation of low-density polyethylene (LDPE) and polystyrene using Bacillus cereus (OR268710) isolated from plastic-polluted tropical coastal environment.

The study focused on marine bacteria, specifically Bacillus cereus, sourced from heavily polluted coastal areas in Tamil Nadu, aiming to assess their efficacy in degrading low-density polyethylene (LDPE) and polystyrene over a 42-day period. When LDPE and polystyrene films were incubated with Bacillus cereus, they exhibited maximum weight losses of 4.13 ± 0.81 % and 14.13 ± 2.41 %, respectively. Notably, polystyrene exhibited a higher reduction rate (0.0036 day-1) and a shorter half-life (195.29 days). SEM images of the treated LDPE and polystyrene unveiled surface erosion with cracks. The energy dispersive X-ray (EDX) analysis revealed elevated carbon content and the presence of oxygen in the treated LDPE and polystyrene films. The ATR-FTIR spectra exhibited distinctive peaks corresponding to functional groups, with observable peak shifts in the treated films. Notable increases were detected in carbonyl, internal double bond, and vinyl indices across all treated groups. Additionally, both treated LDPE and polystyrene showed reduced crystallinity. This research sheds light on Bacillus cereus (OR268710) biodegradation capabilities, emphasizing its potential for eco-friendly waste management in coastal regions.

Biodegradation of polyethylene (PE), polypropylene (PP), and polystyrene (PS) microplastics by floc-forming bacteria, Bacillus cereus strain SHBF2, isolated from a commercial aquafarm.

The ubiquitous proximity of the commonly used microplastic (MP) particles particularly polyethylene (PE), polypropylene (PP), and polystyrene (PS) poses a serious threat to the environment and human health globally. Biological treatment as an environment-friendly approach to counter MP pollution has recent interest when the bio-agent has beneficial functions in their ecosystem. This study aimed to utilize beneficial floc-forming bacteria Bacillus cereus SHBF2 isolated from an aquaculture farm in reducing the MP particles (PE, PP, and PS) from their environment. The bacteria were inoculated for 60 days in a medium containing MP particle as a sole carbon source. On different days of incubation (DOI), the bacterial growth analysis was monitored and the MP particles were harvested to examine their weight loss, surface changes, and alterations in chemical properties. After 60 DOI, the highest weight loss was recorded for PE, 6.87 ± 0.92%, which was further evaluated to daily reduction rate (k), 0.00118 day-1, and half-life (t1/2), 605.08 ± 138.52 days. The OD value (1.74 ± 0.008 Abs.) indicated the higher efficiency of bacteria for PP utilization, and so for the colony formation per define volume (1.04 × 1011 CFU/mL). Biofilm formation, erosions, cracks, and fragments were evident during the observation of the tested MPs using the scanning electron microscope (SEM). The formation of carbonyl and alcohol group due to the oxidation and hydrolysis by SHBF2 strain were confirmed using the Fourier transform infrared spectroscopic (FTIR) analysis. Additionally, the alterations of pH and CO2 evolution from each of the MP type ensures the bacterial activity and mineralization of the MP particles. The findings of this study have confirmed and indicated a higher degree of biodegradation for all of the selected MP particles. B. cereus SHBF2, the floc-forming bacteria used in aquaculture, has demonstrated a great potential for use as an efficient MP-degrading bacterium in the biofloc farming system in the near future to guarantee a sustainable green aquaculture production.

Bacterial screening in Indian coastal regions for efficient polypropylene microplastics biodegradation.

Polypropylene based medical devices significantly increased production and usage in COVID-19 pandemic states, and this material is very resilient in the environment. Thus, more than ever, rapid action is needed to reduce this pollution. This study focuses on the degradation of polypropylene microplastics (PP MPs) by unique marine bacterial strains obtained from the Thoundi (Bacillus tropicus, Bacillus cereus, Stenotrophomonas acidaminiphila, and Brucella pseudintermedia) and Rameshwaram coasts (Bacillus cereus). Those above five bacterial strains were chosen after preliminary screening of their hydrophobicity, biofilm-forming capabilities, and responsiveness to the zone of clearance technique. During the biodegradation process (28 days), the growth, metabolic activity, and viability of these five isolates were all raised. After the post-biodegradation process, the weight loss percentages of the mentioned bacterial strains treated with PP MPs gradually decreased, with values of 51.5 ± 0.5 %, 47.5 ± 0.5 %, 33 ± 1 %, 28.5 ± 0.5 and 35.5 ± 0.5 %, respectively. UV-Vis DRS and SEM analysis confirmed that bacterial strains adhering to MPs cause cracks and cavities on their surface. The degradation of PP MPs can be inferred from alterations in the FT-IR spectrum, specifically in the carbonyl group range of 1100-1700 cm-1, as well as changes in the 1H NMR spectrum, including chemical shift and proton peak pattern alterations. Bacterial strains facilitated the degradation of PP MPs through the secretion of hydrolase-categorized enzymes of protease, lipase, and esterase. The findings of this study indicate that marine bacteria may possess distinctive characteristics that facilitate the degradation of plastic waste and contribute to environmental conservation.

Screening of cellulose-degrading bacteria and optimization of cellulase production from Bacillus cereus A49 through response surface methodology.

Cellulose-degrading microorganisms hold immense significance in utilizing cellulose resources efficiently. The screening of natural cellulase bacteria and the optimization of fermentation conditions are the hot spots of research. This study meticulously screened cellulose-degrading bacteria from mixed soil samples adopting a multi-step approach, encompassing preliminary culture medium screening, Congo red medium-based re-screening, and quantification of cellulase activity across various strains. Particularly, three robust cellulase-producing strains were identified: A24 (MT740356.1 Brevibacillus borstelensis), A49 (MT740358.1 Bacillus cereus), and A61 (MT740357.1 Paenibacillus sp.). For subsequent cultivation experiments, the growth curves of the three obtained isolates were monitored diligently. Additionally, optimal CMCase production conditions were determined, keeping CMCase activity as a key metric, through a series of single-factor experiments: agitation speed, cultivation temperature, unit medium concentration, and inoculum volume. Maximum CMCase production was observed at 150 rpm/37 °C, doubling the unit medium addition, and a 5 mL inoculation volume. Further optimization was conducted using the selected isolate A49 employing response surface methodology. The software model recommended a 2.21fold unit medium addition, 36.11 °C temperature, and 4.91 mL inoculant volume for optimal CMCase production. Consequently, three parallel experiments were conducted based on predicted conditions consistently yielding an average CMCase production activity of 15.63 U/mL, closely aligning with the predicted value of 16.41 U/mL. These findings validated the reliability of the model and demonstrated the effectiveness of optimized CMCase production conditions for isolate A49.

Transport and retention of selected engineered nanoparticles by porous media in the presence of a biofilm.

Column experiments were conducted to investigate the transport of aqueous C60 (aqu-nC60), fullerol, silver nanoparticles (NPs) coated with polyvinylpyrrolidone (Ag-PVP) and stabilized by citrate (Ag-CIT) in biofilm-laden porous media. Gram-negative Pseudomonas aeruginosa (PA) and Gram-positive Bacillus cereus (BC) biofilm-laden glass beads were selected to represent the bacterial interfaces NPs might encounter in the natural aquatic environment. The biomass distribution, extracellular polymeric substances (EPS) components, electrokinetic property, and hydrophobicity of these interfaces were characterized, and the hydrophobicity was found to correlate with the quantity of proteins in EPS. The retention of NPs on glass beads coated with bovine serum albumin (BSA) and alginate were also studied. Except for Ag-PVP, the affinity of NPs for porous medium, indicated by attachment efficiency α, increased in the presence of biofilms, BSA and alginate. For hydrophobic aqu-nC60, the larger the proteins/polysaccharides ratio, the larger the α, suggesting the hydrophobic interaction determines the attachment of aqu-nC60 to the collector surface. Uncharged PVP stabilized Ag-PVP by steric repulsion, and the attachment to glass beads was not enhanced by biofilm. The presence of divalent ion Ca(2+) significantly hydrophobized biofilm, BSA, and alginate-coated glass beads and further retarded the mobility of aqu-nC60, fullerol, and Ag-CIT; while Ag-PVP was again sterically stabilized.

Synthesis of poly-(3-hydroxybutyrate-co-12 mol % 3-hydroxyvalerate) by Bacillus cereus FB11: its characterization and application as a drug carrier.

The synthesis of microbial polyhydroxyalkanoate is investigated in this work for it potential application as drug carrier for cancer therapy. The bacterial isolate Bacillus cereus FB11 has synthesized poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer under nutrient stress conditions using glucose as a sole carbon source. The FTIR spectrum of the purified copolymer showed the characteristic absorption bands at 1,719, 1,260 and 2,931 cm(-1) attributing to C=O, C-O stretching and C-H vibrations, respectively. The result of (1)H-NMR confirmed that it was composed of 88 mol % of 3-hydroxybutyrate and 12 mol % of 3-hydroxyvalerate monomeric subunits. The nanoparticles were fabricated from copolymer and used as a carrier for anticancer drug ellipticine. The in vitro drug release studies showed that % inhibition of A549 cancer cell line receiving ellipticine loaded poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) nanoparticles was two-fold higher in comparison to ellipticine alone. This drug delivery system offers exciting possibilities for cancer therapy by increasing the bioavailability of anti-neoplastic drug to the tumor site.

The escalated potential of the novel isolate Bacillus cereus NJD1 for effective biodegradation of LDPE films without pre-treatment.

This study focused on the biodegradation of LDPE films using a novel isolate of Bacillus obtained from soil samples collected from a 20-year-old plastic waste dump. The aim was to evaluate the biodegradability of LDPE films treated with this bacterial isolate. The results indicated a 43% weight loss of LDPE films within 120 days of treatment. The biodegradability of LDPE films was confirmed through various testing methods, including BATH, FDA, CO2 evolution tests, and changes in total cell growth count, protein content, viability, pH of the medium, and release of microplastics. The bacterial enzymes, including laccases, lipases, and proteases, were also identified. SEM analysis revealed biofilm formation and surface changes in treated LDPE films, while EDAX analysis showed a reduction in carbon elements. AFM analysis demonstrated differences in roughness compared to the control. Furthermore, wettability increased and tensile strength decreased, confirming the biodegradation of the isolate. FTIR spectral analysis showed changes in skeletal vibrations, such as stretches and bends, in the linear structure of polyethylene. FTIR imaging and GC-MS analysis also confirmed the biodegradation of LDPE films by the novel isolate identified as Bacillus cereus strain NJD1. The study highlights the potentiality of the bacterial isolate for safe and effective microbial remediation of LDPE films.

Genome-centric polyhydroxyalkanoate reconciliation reveals nutrient enriched growth dependent biosynthesis in Bacillus cereus IBA1.

Microbiological polyhydroxyalkanoates (PHAs) are rooted as the most promising bio-replacements of synthetic polymers. Inherent properties of these PHAs further expand their applicability in numerous industrial, environmental, and clinical sectors. To propel these, a new environmental, endotoxin free gram-positive bacterium i.e., Bacillus cereus IBA1 was identified to harbor advantageous PHA producer characteristics through high-throughput omics mining approaches. Unlike traditional fermentations, nutrient enriched strategy was used to enhance PHA granular concentrations by ∼2.3 folds to 2.78 ± 0.19 g/L. Additionally, this study is the first to confirm an underlying growth dependent PHA biogenesis through exploring PHA granule associated operons which harbour constitutively expressing PHA synthase (phaC) coupled with differentially expressing PHA synthase subunit (phaR) and regulatory protein (phaP, phaQ) amid different growth phases. Moreover, the feasibility of this promising microbial phenomenon could propel next-generation biopolymers, and increase industrial applicability of PHAs, thereby significantly contributing to the sustainable development.

Response surface optimization of poly-ß-hydroxybutyrate synthesized by Bacillus cereus L17 using acetic acid as carbon source.

A strain of Bacillus that can tolerate 10 g/L acetic acid and use the volatile fatty acids produced by the hydrolysis and acidification of activated sludge to produce polyhydroxyalkanoate was screened from the activated sludge of propylene oxide saponification wastewater. The strain was identified by 16S rRNA sequencing and phylogenetic tree analysis and was named Bacillus cereus L17. Various characterization methods showed that the polymer synthesized by strain L17 is poly-ß-hydroxybutyrate, which has low crystallinity, good ductility and toughness, high thermal stability and a low polydispersity coefficient. It has wide thermoplastic material operating space as well as industrial and medicinal applications. The optimal fermentation conditions were determined by single factor optimization. Then, Plackett-Burman and Box-Behnken design experiments were carried out according to the single factor optimization results, and the response surface optimization was completed. The final results were: initial pH 6.7, temperature 25 °C, and loading volume 124 mL. The verification experiment showed that the yield of poly-ß-hydroxybutyrate after optimization increased by 35.2 % compared to that before optimization.

Atomic structure of a Na+- and K+-conducting channel.

Ion selectivity is one of the basic properties that define an ion channel. Most tetrameric cation channels, which include the K+, Ca2+, Na+ and cyclic nucleotide-gated channels, probably share a similar overall architecture in their ion-conduction pore, but the structural details that determine ion selection are different. Although K+ channel selectivity has been well studied from a structural perspective, little is known about the structure of other cation channels. Here we present crystal structures of the NaK channel from Bacillus cereus, a non-selective tetrameric cation channel, in its Na+- and K+-bound states at 2.4 A and 2.8 A resolution, respectively. The NaK channel shares high sequence homology and a similar overall structure with the bacterial KcsA K+ channel, but its selectivity filter adopts a different architecture. Unlike a K+ channel selectivity filter, which contains four equivalent K+-binding sites, the selectivity filter of the NaK channel preserves the two cation-binding sites equivalent to sites 3 and 4 of a K+ channel, whereas the region corresponding to sites 1 and 2 of a K+ channel becomes a vestibule in which ions can diffuse but not bind specifically. Functional analysis using an 86Rb flux assay shows that the NaK channel can conduct both Na+ and K+ ions. We conclude that the sequence of the NaK selectivity filter resembles that of a cyclic nucleotide-gated channel and its structure may represent that of a cyclic nucleotide-gated channel pore.