Microbial-based metabolites associated with degradation of imidacloprid and its impact on stress-responsive proteins.

Imidacloprid (IMD), a neonicotinoid insecticide, is intensively used in agricultural fields for effective protection against aphids, cane beetles, thrips, stink bugs, locusts, etc., is causing serious environmental concerns. In recent years, seed treatment with Imidacloprid is being practiced mainly to prevent sucking insect pests. In India, due to the increase in application of this insecticide residue has been proven to have an impact on the quality of soil and water. In view of this, the current investigation is focussed on sustainable approach to minimize the residual effect of IMD in agricultural fields. The present study reveals a most promising imidacloprid resistant bacterium Lysinibacillus fusiformis IMD-Bio5 strain isolated from insecticide-contaminated soil. The isolated bacterial strain upon tested for its biodegradation potential on mineral salt medium (MSM) showed a significant survival growth at 150 g/L of IMD achieved after 3 days, whereas immobilized cells on MSM amended with 200 g/L of IMD as the sole carbon source provided degradation of 188 and 180 g/L of IMD in silica beads and sponge matrices, respectively. The liquid chromatography mass spectrometry was performed to test the metabolite responsive for IMD biodegradation potential of L. fusiformis IMD-Bio5 which showed the induced activity of the metabolite 6-Chloronicotinic acid. Furthermore, as compared to the untreated control, the Lysinibacillus fusiformis IMD-Bio5 protein profile revealed a range of patterns showing the expression of stress enzymes. Thus, results provided a most effective bacterium enabling the removal of IMD-like hazardous contaminants from the environment, which contributes to better agricultural production and soil quality, while long-term environmental advantages are restored.

Lysinibacillus fusiformis bacteremia: Case report and literature review.

A 93-year-old woman was diagnosed with Lysinibacillus fusiformis bacteremia complicated with coma blisters. Initial gram staining for L. fusiformis indicated the presence of gram-negative rods; however, subsequent staining of colonies from Mueller-Hinton agar revealed the presence of gram-positive and gram-negative rods with spherical endospores, and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (VITEK ® MS and microflex® LT/SH) definitively identified the organism as L. fusiformis. The two-week administration of piperacillin/tazobactam and ampicillin resulted in an improvement of the patient's general condition, and the skin lesions gradually improved.

Transcriptome analysis provides new insights into the tolerance and reduction of Lysinibacillus fusiformis 15-4 to hexavalent chromium.

Microbial bioremediation of Cr(VI)-contaminated environments has drawn extensive concern. However, the molecular processes underlying the microbial Cr(VI) tolerance and reduction remain unclear. We isolated a Cr(VI)-reducing Lysinibacillus fusiformis strain 15-4 from soil on the Qinghai-Tibet Plateau. When grown in 1 mM and 2 mM Cr(VI)-containing medium, strain 15-4 could reduce 100% and 93.7% of Cr(VI) to Cr(III) after 36 h and 60 h of incubation, respectively. To know the molecular processes in response to Cr(VI), transcriptome sequencing was carried out using RNA-Seq technology. The results annotated a total of 3913 expressed genes in the strain. One thousand ninety-eight genes (28.1%) were significantly (fold change ≥ 2, false discovery rate ≤ 0.05) expressed in response to Cr(VI), of which 605 (55.1%) were upregulated and 493 (44.9%) were downregulated. The enrichment analysis showed that a total of 630 differentially expressed genes (DEGs) were enriched to 122 KEGG pathways, of which 8 pathways were significantly (p < 0.05) enriched in Cr(VI)-treated sample, including ATP-binding cassette (ABC) transporters (97 DEGs), ribosome (40), sulfur metabolism (16), aminoacyl-tRNA biosynthesis (19), porphyrin metabolism (20), quorum sensing (44), oxidative phosphorylation (17), and histidine metabolism (10), suggesting that these pathways play key roles to cope with Cr(VI) in the strain. The highly upregulated DEGs consisted of 29 oxidoreductase, 18 dehydrogenase, 14 cell redox homeostasis and stress response protein, and 10 DNA damage and repair protein genes. However, seven Na+:H+ antiporter complex-coding DEGs and most of transcriptional regulator-coding DEGs were significantly downregulated in the Cr-treated sample. Many of FMN/NAD(P)H-dependent reductase-encoding genes were greatly induced by Cr, suggesting the involvement of these genes in Cr(VI) reduction in strain 15-4. Sulfur and iron ions as well as the thiol-disulfide exchange reactions might play synergistic roles in Cr reduction.Key points• Lysinibacillus fusiformis 15-4 was able to tolerate and reduce Cr(VI) to Cr(III).• Transcriptome analysis revealed that 1098 DEGs and 8 key KEGG pathways significantly responded to Cr(VI).• Sulfur metabolism, protein biosynthesis, and porphyrin metabolism were the key pathways associated with the survival of strain 15-4 in response to Cr(VI).

Effect of milk bactofugation on the counts and diversity of thermoduric bacteria.

The objective of this work was to determine the effect of milk bactofugation on the counts and microbial diversity of mesophilic (MT), psychrotrophic (PT), and thermophilic (TT) thermoduric bacteria and its potential as a technological method to remove spoilage microorganisms resistant to pasteurization. Different batches of raw milk from 69 dairy farms divided into sets in 3 bulk tanks (A, B, C) were evaluated at different times during the technological process. As the raw milk was preheated (∼55°C) immediately before bactofugation (10,000 × g), the effect of bactofugation was estimated by comparing the counts in raw, preheated, and bactofuged milk. This centrifugation was sufficient to reduce the isolation of 88% of the MT in preheated milk. For PT, it was possible to verify a reduction of 72.5% in batch C. The TT were not recovered at higher detection limits (<5 cfu/mL). For diversity, 310 isolates were identified using a molecular approach; 15 species of contaminating thermoduric bacteria were identified from raw and preheated milk, and only 6 species were recovered in bactofuged milk. Only MT were recovered from the bactofuged milk, mainly the species Lysinibacillus fusiformis (61.7%) and Bacillus licheniformis (12.3%). Both species are known to be endospore-forming psychrotrophs and have proteolytic or lipolytic activity. The bactofugation of raw milk reduced the number of isolates of B. licheniformis, Bacillus toyonensis, Micrococcus aloeverae, and Aestuariimicrobium kwangyangense by 33, 43, 86, and 92%, respectively, and reduced the isolates of Macrococcus caseolyticus, Lysinibacillus varians, Carnobacterium divergens, Microbacterium hominis, Kocuria indica, Micrococcus yunnanensis, Gordonia paraffinivorans, Bacillus invictae, and Kocuria kristinae to undetectable levels. The results of this study indicate that bactofugation can be applied by the dairy industry to reduce pasteurization-resistant microorganisms in combination with prophylactic measures to prevent the contamination of raw milk by spores and vegetative forms of bacteria.

Transcriptome profiling reveals the molecular processes for survival of Lysinibacillus fusiformis strain 15-4 in petroleum environments.

A bacterial strain designated Lysinibacillus fusiformis 15-4 was isolated from oil-free soil on the Qinghai-Tibet Plateau, which can grow well utilizing petroleum hydrocarbons as a carbon source at a lower temperature. To deeply characterize the molecular adaptations and metabolic processes of this strain when grown in a petroleum-containing environment, transcriptome analysis was performed. A total of 4664 genes and the expression of 3969 genes were observed in strain 15-4. When the strain was grown in petroleum-containing medium, 2192 genes were significantly regulated, of which 1312 (60%) were upregulated and 880 (40%) were downregulated. This strain degraded and adapted to petroleum via modulation of diverse molecular processes, including improvements in transporter activity, oxidoreductase/dehydrogenase activity, two-component system/signal transduction, transcriptional regulation, fatty acid catabolism, amino acid metabolism, and environmental stress responses. Many strain-specific genes were involved in the oxidation of hydrocarbon compounds, such as several luciferase family alkane monooxygenase genes, flavin-utilizing monooxygenase family genes, and flavoprotein-like family alkanesulfonate monooxygenase genes. Several cold shock protein genes were also induced suggesting adaptation to cold environments and the potential for petroleum degradation at low temperatures. The results obtained in this study may broaden our understanding of molecular adaptation of bacteria to hydrocarbon-containing environments and may provide valuable data for further study of L. fusiformis.

Secondary metabolites of Bacillus subtilis impact the assembly of soil-derived semisynthetic bacterial communities.

Secondary metabolites provide Bacillus subtilis with increased competitiveness towards other microorganisms. In particular, nonribosomal peptides (NRPs) have an enormous antimicrobial potential by causing cell lysis, perforation of fungal membranes, enzyme inhibition, or disruption of bacterial protein synthesis. This knowledge was primarily acquired in vitro when B. subtilis was competing with other microbial monocultures. However, our understanding of the true ecological role of these small molecules is limited. In this study, we have established soil-derived semisynthetic mock communities containing 13 main genera and supplemented them with B. subtilis P5_B1 WT, the NRP-deficient strain sfp, or single-NRP mutants incapable of producing surfactin, plipastatin, or bacillaene. Through 16S amplicon sequencing, it was revealed that the invasion of NRP-producing B. subtilis strains had no major impact on the bacterial communities. Still, the abundance of the two genera Lysinibacillus and Viridibacillus was reduced. Interestingly, this effect was diminished in communities supplemented with the NRP-deficient strain. Growth profiling of Lysinibacillus fusiformis M5 exposed to either spent media of the B. subtilis strains or pure surfactin indicated the sensitivity of this strain towards the biosurfactant surfactin. Our study provides a more in-depth insight into the influence of B. subtilis NRPs on semisynthetic bacterial communities and helps to understand their ecological role.

Lysinibacillus fusiformis M5 Induces Increased Complexity in Bacillus subtilis 168 Colony Biofilms via Hypoxanthine.

In recent years, biofilms have become a central subject of research in the fields of microbiology, medicine, agriculture, and systems biology, among others. The sociomicrobiology of multispecies biofilms, however, is still poorly understood. Here, we report a screening system that allowed us to identify soil bacteria which induce architectural changes in biofilm colonies when cocultured with Bacillus subtilis We identified the soil bacterium Lysinibacillus fusiformis M5 as an inducer of wrinkle formation in B. subtilis colonies mediated by a diffusible signaling molecule. This compound was isolated by bioassay-guided chromatographic fractionation. The elicitor was identified to be the purine hypoxanthine using mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. We show that the induction of wrinkle formation by hypoxanthine is not dependent on signal recognition by the histidine kinases KinA, KinB, KinC, and KinD, which are generally involved in phosphorylation of the master regulator Spo0A. Likewise, we show that hypoxanthine signaling does not induce the expression of biofilm matrix-related operons epsABCDEFGHIJKLMNO and tasA-sipW-tapA Finally, we demonstrate that the purine permease PbuO, but not PbuG, is necessary for hypoxanthine to induce an increase in wrinkle formation of B. subtilis biofilm colonies. Our results suggest that hypoxanthine-stimulated wrinkle development is not due to a direct induction of biofilm-related gene expression but rather is caused by the excess of hypoxanthine within B. subtilis cells, which may lead to cell stress and death.IMPORTANCE Biofilms are a bacterial lifestyle with high relevance regarding diverse human activities. Biofilms can be beneficial, for instance, in crop protection. In nature, biofilms are commonly found as multispecies communities displaying complex social behaviors and characteristics. The study of interspecies interactions will thus lead to a better understanding and use of biofilms as they occur outside laboratory conditions. Here, we present a screening method suitable for the identification of multispecies interactions and showcase L. fusiformis as a soil bacterium that is able to live alongside B. subtilis and modify the architecture of its biofilms.

Highly luminescent and biocompatible, L-citrulline-capped ZnS:Mn nanocrystals for rapid screening of metal accumulating Lysinibacillus fusiformis bacteria.

Biocompatible and highly luminescent manganese doped zinc sulfide (ZnS:Mn) nanocrystals of average particle size 10 nm have been synthesized by capping with a novel amino acid ligand, L-citrulline. Though there are many reports on the bioimaging applications of nanostructured semiconductors, the present study focused on the detection of a special type of metal accumulating bacteria, Lysinibacillus fusiformis. This bacterium has significant applications in the disposal of metal components from industrial effluents. In this context, the detection of this bacterium is quite important and the present work demonstrates a novel technique for this bacterial detection. The synthesized nanocrystals were attached to Lysinibacillus fusiformis and characteristics of the bioconjugated system were studied. The blue shift observed in the ultraviolet-visible absorption and photoluminescence spectra of the bioconjugated system, confirms conjugation of the Lysinibacillus fusiformis with L-citrulline-capped ZnS:Mn. When the bioconjugated system (capped ZnS:Mn + bacteria) was observed using a fluorescent microscope under excitation wavelengths 365.4 nm (ultraviolet), 435.8 nm (blue) and 546.1 nm (green), fluorescence emissions were obtained in yellow, green and red regions respectively. The study of relative growth of Lysinibacillus fusiformis in the presence of L-citrulline-capped ZnS:Mn proves biocompatible property of these nanocrystals and their tunable color properties under different excitation wavelengths make them ideal for biolabeling applications.

Purification and characterization of an alkali-organic solvent-stable laccase with dye decolorization capacity from newly isolated Lysinibacillus fusiformis W11.

A new Lysinibacillus fusiformis strain with abundant laccase activity was isolated from soil under forest rotted leaf and identified as L. fusiformis W11 based on its 16S rRNA gene sequence and physiological characteristics. The laccase LfuLac was purified and characterized. The optimum temperature and pH of LfuLac on guaiacol were 45 °C and pH 9, respectively. LfuLac kept 78%, 88%, 92%, 74%, and 47% of activity at pH 7-11, respectively, suggesting the alkali resistance of the enzyme. The effects of various metal ions on LfuLac showed that Cu2+, Mg2+, and Na+ were beneficial to laccase activity and 10 mM Cu2+ increased the activity of LfuLac to 216%. LfuLac showed about 90% activity at 5% organic solvents and more than 60% activity at 20%, indicating its resistance to organic solvents. In addition, LfuLac decolorized different kinds of dyes. This study enriched our knowledge about laccase from L. fusiformis W11 and its potential industrial applications.

Simultaneous nitrogen removal via heterotrophic nitrification and aerobic denitrification by a novel Lysinibacillus fusiformis B301.

Simultaneous nitrogen removal via heterotrophic nitrification and aerobic denitrification (HN-AD) has received widespread attention in biological treatment of wastewater. This study reported a novel Lysinibacillus fusiformis B301 strain, which effectively removed nitrogenous pollutants via HN-AD in one aerobic reactor with no nitrite accumulated. It exhibited the optimal nitrogen removal efficiency under 30°C, citrate as the carbon source and C/N ratio of 15. The maximum nitrogen removal rates were up to 2.11 mgNH4 + -N/(L·h), 1.62 mgNO3 - -N/(L·h), and 1.41 mgNO2 - -N/(L·h), respectively, when ammonium, nitrate, and nitrite were employed as the only nitrogen source under aerobic conditions. Ammonium nitrogen was preferentially consumed via HN-AD in the coexistence of three nitrogen species, and the removal efficiencies of total nitrogen were up to 94.26%. Nitrogen balance analysis suggested that 83.25% of ammonium was converted to gaseous nitrogen. The HD-AD pathway catalyzed by L. fusiformis B301 followed NH 4 + → N H 2 OH → NO 2 - → NO 3 - → NO 2 - → N 2 , supported by the results of key denitrifying enzymatic activities. PRACTITIONER POINTS: The novel Lysinibacillus fusiformis B301 exhibited the outstanding HN-AD ability. The novel Lysinibacillus fusiformis B301 simultaneously removed multiple nitrogen species. No nitrite accumulated during the HN-AD process. Five key denitrifying enzymes were involved in the HN-AD process. Ammonium nitrogen (83.25%) was converted to gaseous nitrogen by the novel strain.

The effect of BVOCs produced by Lysinibacillus fusiformis and LED irradiation on pigment metabolism in stored broccoli.

Volatile organic compounds produced by bacteria (BVOCs) have been proven to effect the postharvest metabolism of fruits and vegetables. The quality, color and antioxidant capacity of membrane lipids of broccoli in storage were effectively maintained by fumigation with BVOCs produced by Lysinibacillus fusiformis combined with white light emitting diode (LED) technology. An analysis of the transcriptome and metabolome of broccoli treated with the combined LED-BVOCs technology resulted in the identification of 49 differentially expressed genes (DEGs) and 13 differentially abundant metabolites (DAMs) involved in photosynthesis (32/0 DEGs upregulated/downregulated; 0/0 DAMs with increased/decreased abundance), chlorophyll (7/0; 1/2), carotenoid (5/0; 1/4) and flavonoid (3/3; 3/2) metabolism. The maintenance of green color in harvested broccoli treated by LED-BVOCs was associated with DEGs and DAMs that inhibited chlorophyll degradation and carotenoid accumulation. Our study provides a theoretical basis for understanding the delayed senescence of broccoli during storage using BVOCs-LED technology.

Production and biochemical characterization of partially purified cellulase-free, thermo-acidophilic endoxylanase from Lysinibacillus fusiformis strain TB7 using kolanut husk as feedstock.

Xylanases have become very important enzymes in many industrial processes for the valorization of xylan-rich lignocellulosic wastes. Here, some physicochemical and kinetic properties of a purified endoxylanase produced on kolanut husk-based medium by Lysinibacillus fusiformis are presented. The crude enzyme solution was first subjected to precipitation with solid ammonium sulphate and further purified on DEAE-Sephadex A-50 anion-exchange and Sephadex G-100 gel filtration columns chromatography prior to biochemical characterization. The purified endoxylanase was 21 kDa as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and was thermostable, exhibiting optimum activity at 60 °C and pH 5.0. The K m and V max were respectively estimated to be 29.5 mg/ml and 125 µmol/min/ml using Birchwood xylan as substrate. Activity of the enzyme was enhanced by Na+, Ca2+, Mn2+, Mg2+ and K+ at concentration of 5 mM but inhibited by Hg2+, Cu2+, Pb2+, Fe3+, EDTA, SDS and Urea. The purified endoxylanase showed high hydrolytic activity on Birchwood xylan and kolanut husk but extremely poor or no activity on carboxymethyl cellulose, starch or pectin. This L. fusiformis strain TB7 endoxylanase has desirable properties useful for biotechnological applications in laundry, fuels, feeds, paper and pulp industries.

Biodegradation of Congo red dye using polyurethane foam-based biocarrier combined with activated carbon and sodium alginate: Batch and continuous study.

Dyes are an important class of organic pollutants and are well known for their adverse effects on aquatic life and human beings. In this work, an effort has been made to treat the dye-containing wastewater using modified biocarriers in packed bed bioreactors (PBBRs). Lysinibacillus sp. immobilized polyurethane foam combined with activated carbon and sodium alginate was used for the biodegradation of Congo red dye. The optimum values of process time, glucose concentration, and dye concentration were obtained to be 4.0 days, 2.0 g/L, and 50 mg/L, respectively. The maximum dye removal efficiency (RE) of 92.63 % was obtained at the optimized conditions. The continuous PBBR offered a maximum RE and elimination capacity of 90.73% and 10.89 mg/L. d, respectively, at an inlet loading rate of 12 mg/L. d. Moreover, the growth kinetic of Lysinibacillus sp. was well predicted by the Andrew-Haldane model with a regression coefficient of 0.98.

Evaluation of biosurfactant production potential of Lysinibacillus fusiformis MK559526 isolated from automobile-mechanic-workshop soil.

BACKGROUND: Biosurfactants are amphipathic biological compounds with surface active potential and are produced by many microorganisms. Biosurfactant production by Lysinibacillus fusiformis MK559526 isolated from automobile-mechanic-shop soil was investigated with a view to assessing its potential for production and potential for optimization. MATERIALS AND METHODS: Effects of carbon and nitrogen sources, pH, temperature and incubation periods on biosurfactant production were evaluated with a view to optimizing the processes. Fourier Transform Infra-Red absorption peaks and Gas chromatography mass spectrometry were used to determine the functional groups of the chemical make-up and the chemical profile of the biosurfactant respectively. RESULTS: Lysinibacillus fusiformis surfactant had emulsification index of 65.15 ± 0.35 %, oil displacement of 2.7 ± 0.26 mm, zone of haemolysis of 7.3 ± 0.16 mm and a positive drop collapse test. Optimized culture conditions for biosurfactant production: temperature, 35 ºC; pH, 7.0; starch solution, 40 g/L and urea, 1.5 g/L showed a reduction in surface tension to 28.46 ± 1.11 mN/m and increased emulsification index to 93.80 ± 0.41 %. Maximum biosurfactant production of 2.92 ± 0.04 g/L was obtained after 72 h. The biosurfactant contained peptides and fatty acids. The predominant fatty acid was 9-Octadecenoic acid (80.80%). CONCLUSIONS: The above results showing high emulsification potential and remarkable reduction in the surface tension are good biosurfactant attributes. Consequently, Lysinibacillus fusiformis MK559526 is a good candidate for biosurfactant production.

Characterization of Lysinibacillus fusiformis strain S4C11: In vitro, in planta, and in silico analyses reveal a plant-beneficial microbe.

Despite sharing many of the traits that have allowed the genus Bacillus to gain recognition for its agricultural relevance, the genus Lysinibacillus is not as well-known and studied. The present study employs in vitro, in vivo, in planta, and in silico approaches to characterize Lysinibacillus fusiformis strain S4C11, isolated from the roots of an apple tree in northern Italy. The in vitro and in vivo assays demonstrated that strain S4C11 possesses an antifungal activity against different fungal pathogens, and is capable of interfering with the germination of Botrytis cinerea conidia, as well as of inhibiting its growth through the production of volatile organic molecules. In planta assays showed that the strain possesses the ability to promote plant growth, that is not host-specific, both in controlled conditions and in a commercial nursery. Biocontrol assays carried out against phytopathogenic viruses gave contrasting results, suggesting that the strain does not activate the host's defense pathways. The in silico analyses were carried out by sequencing the genome of the strain through an innovative approach that combines Illumina and High-Definition Mapping methods, allowing the reconstruction of a main chromosome and two plasmids from strain S4C11. The analysis of the genes encoded by the genome contributed to the characterization of the strain, detecting genes related to the biocontrol effect detected in the experimental trials.

Optimization of glutaminase-free L-asparaginase production using mangrove endophytic Lysinibacillus fusiformis B27.

Background: The mangrove, Rhizophora mucronata, an essential source of endophytic bacteria, was investigated for its ability to produce glutaminase-free L-asparaginase. The study aimed to obtain glutaminase-free L-asparaginase-producing endophytic bacteria from the mangrove and to optimize enzyme production. Methods: The screening of L-asparaginase-producing bacteria used modified M9 medium. The potential producer was further analyzed with respect to its species using 16S rRNA gene sequencing. Taguchi experimental design was applied to optimize the enzyme production. Four factors (L-asparagine concentration, pH, temperature, and inoculum concentration) were selected at four levels. Results: The results indicated that the endophytic bacteria Lysinibacillus fusiformis B27 isolated from R. mucronata was a potential producer of glutaminase-free L-asparaginase. The experiment indicated that pH 6, temperature at 35°C, and inoculum concentration of 1.5% enabled the best production and were essential factors. L-asparagine (2%) was less critical for optimum production. Conclusions: L. fusiformis B27, isolated from Rhizophora mucronata, can be optimized for L-ASNase enzyme production using optimization factors (L-ASNase, pH, temperature, and inoculum), which can increase L-ASNase enzyme production by approximately three-fold.

Highly efficient deproteinization with an ammonifying bacteria Lysinibacillus fusiformis isolated from brewery spent diatomite.

To explore a new method for bio-regeneration of high-protein brewery spent diatomite, an ammonifying bacteria (BSD1) was screened out from it and identified as Lysinibacillus fusiformis. The protein degradation characteristics of BSD1 was studied with rice protein as the sole nitrogen source. Maximum protein degradation activity was obtained when BSD1 was inoculated with an inoculum of 5% into a medium with glucose as carbon source and initial pH value of 7.0 and incubated at 30°C for 48 h. In this optimal condition, protein concentration decreased from 156.8 mg/L to 19.2 mg/L, and protein degradation efficiency of BSD1 reached 88%. Free amino acid analysis showed that the content of Phe, Tyr, Pro, Ala, Lys, Thr and His increased in protein degradation process. After degradation, NH4+N concentration producing in medium supernatant reached 232.2 mg/L. These results indicated the strain BSD1 could transform proteins into free amino acids and eventually convert them to ammonium or ammonia. Furthermore, strain BSD1 could also be used for deproteinization of brewery spent diatomite and 51% of proteins in spent diatomite were degraded. After biological deproteinization the specific surface area and total pore volume of diatomite improved obviously. These results illustrated that the application of strain BSD1 for bio-regeneration of high-protein brewery spent diatomite was efficient and feasible.

Draft genome sequence data of Lysinibacillus fusiformis strain GM, isolated from potato phyllosphere as a potential probiotic.

Here we present the morphological and physiological properties of isolated Lysinibacillus fusiformis strain GM, its draft genome sequence as well as annotation and analysis of its genome. Initial analysis of MALDI-TOF mass spectrometry, 16S rRNA gene analysis and in silico DNA-DNA hybridization revealed that the strain belongs to the species Lysinibacillus fusiformis. The 4,678,122 bp draft genome consist of 17 scaffolds encoding 4588 proteins and 137 RNAs. Annotation of the genome sequence revealed cellulase and protease encoding genes, genes of adhesion proteins and putative genes responsible for the biosynthesis of antimicrobial metabolites. The Whole Genome Shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession number NTMQ00000000.1 (https://www.ncbi.nlm.nih.gov/nuccore/NZ_NTMQ00000000.1).

Optimized production and characterization of a detergent-stable protease from Lysinibacillus fusiformis C250R.

In this study, we aimed to optimize the cultural and nutritional conditions for protease production by Lysinibacillus fusiformis strain C250R in submerged fermentation process using statistical methodology. The most significant factors (gruel, wheat bran, yeast extract, and FeSO4) were identified by Plackett-Burman design. Response surface methodology (RSM) was used to determine the optimum levels of the screened factors and their interaction. Under the optimized conditions, protease yield 3100U/mL was 4.5 folds higher than those obtained by the use of the initial conditions (680U/mL). Additionally, a new extracellular 51kDa-protease, designated SAPLF, was purified and biochemically characterized from strain C250R. It shows optimum activity at 70°C and pH 10. Its half-life times at 70 and 80°C were 10 and 6-h, respectively. Irreversible inhibition of enzyme activity of SAPLF with serine protease inhibitors demonstrated that it belongs to the serine protease family. Interestingly, its catalytic efficiency was higher than that of SPVP from Aeribacillus pallidus strain VP3 and Alcalase Ultra 2.5L from Bacillus licheniformis. This study demonstrated that SAPLF has a high detergent compatibility and an excellent stain removal compared to Alcalase Ultra 2.5L; which offers an interesting potential for its application in the laundry detergent industry.

Production of a microcapsule agent of chromate-reducing Lysinibacillus fusiformis ZC1 and its application in remediation of chromate-spiked soil.

Lysinibacillus fusiformis ZC1 is an efficient Cr(VI)-reducing bacterium that can transform the toxic and soluble chromate [Cr(VI)] form to the less toxic and precipitated chromite form [Cr(III)]. As such, this strain might be applicable for bioremediation of Cr(VI) in soil by reducing its bioavailability. The study objective was to prepare a microcapsule agent of strain ZC1 for bioremediation of Cr(VI)-contaminated soil. Using a single-factor orthogonal array design, the optimal fermentation medium was obtained and consisted of 6 g/L corn flour, 12 g/L soybean flour, 8 g/L NH4Cl and 6 g/L CaCl2. After enlarged fermentation, the cell and spore densities were 5.9 × 10(9) and 1.7 × 10(8) cfu/mL, respectively. The fermentation products were collected and embedded with 1 % gum arabic and 1 % sorbitol as the microcapsule carriers and were subsequently spray-dried. Strain ZC1 exhibited viable cell counts of (3.6 ± 0.44) × 10(10) cfu/g dw after 50-day storage at room temperature. In simulated soil bioremediation experiments, 67 % of Cr(VI) was reduced in 5 days with the inoculation of this microcapsule agent, and the Cr(VI) concentration was below the soil Cr(VI) standard level. The results demonstrated that the microcapsule agent of strain ZC1 is efficient for bioremediation of Cr(VI)-contaminated soil.