Compatibility testing and enhancing the pulp bleaching process by hydrolases of the newly isolated thermophilic <i>Isoptericola variabilis</i> strain UD-6

In the present study, Isoptericola variabilis strain UD-6 isolated from alkaline hot spring of Unapdev, Maharashtra, India was assessed for its biobleaching activity by hydrolytic enzymes on rice straw pulp. Results of primary and secondary screening manifested that it was a multi-enzyme producer, competent to produce amylase, cellulase, mannanase, pectinase, and xylanase at 9.73, 4.11, 6.26, 8.42, and 6.61 IU ml−1 in fermentation conditions, respectively. Maximum activity of all enzymes was gained at thermal temperature (50–55 °C), alkaline condition (pH 8–9), under 5 mM KCl and 5 mM NaCl salt concentration. In compatibility testing, activities of all enzymes were spectacularly reduced when they utilized with chemicals of pulp bleaching. Results of rice straw pulp bleaching was effectual when pulp was initially bleached with mannanase, pectinase, and xylanase enzymes (Es) for 90 min and then with diluted chemicals (DC) for further 90 min instead of their separate use. Treatment of rice straw pulp with Es + DC, enhanced the release of reducing sugars, hydrophobic compounds, and phenolic compounds, whereas Kappa number was reduced. Overall, the results of the present study indicated that pre-bleaching of pulp with hydrolytic enzymes obtained from I. variabilis strain UD-6 helps to minimize chemicals used in the bleaching process and make it more sustainable for pulp and paper industries as well as for the environment.</p

R software package based statistical optimization of process components to simultaneously enhance the bacterial growth, laccase production and textile dye decolorization with cytotoxicity study.

The thermophilic bacterium, Bacillus licheniformis U1 is used for the optimization of bacterial growth (R1), laccase production (R2) and synthetic disperse blue DBR textile dye decolorization (R3) in the present study. Preliminary optimization has been performed by one variable at time (OVAT) approach using four media components viz., dye concentration, copper sulphate concentration, pH, and inoculum size. Based on OVAT result further statistical optimization of R1, R2 and R3 performed by Box-Behnken design (BBD) using response surface methodology (RSM) in R software with R Commander package. The total 29 experimental runs conducted in the experimental design study towards the construction of a quadratic model. The model indicated that dye concentration 110 ppm, copper sulphate 0.2 mM, pH 7.5 and inoculum size 6% v/v were found to be optimum to maximize the laccase production and bacterial growth. Whereas, maximum dye decolorization achieved in media containing dye concentration 110 ppm, copper sulphate 0.6 mM, pH 6 and inoculum size 6% v/v. R package predicted R2 of R1, R2 and R3 were 0.9917, 0.9831 and 0.9703 respectively; likened to Design-Expert (Stat-Ease) (DOE) predicted R2 of R1, R2, and R3 were 0.9893, 0.9822 and 0.8442 respectively. The values obtained by R software were more precise, reliable and reproducible, compared to the DOE model. The laccase production was 1.80 fold increased, and 2.24 fold enhancement in dye decolorization was achieved using optimized medium than initial experiments. Moreover, the laccase-treated sample demonstrated the less cytotoxic effect on L132 and MCF-7 cell lines compared to untreated sample using MTT assay. Higher cell viability and lower cytotoxicity observed in a laccase-treated sample suggest the impending application of bacterial laccase in the reduction of toxicity of dye to design rapid biodegradation process

Metagenomic data of fungal internal transcribed Spacer and 18S rRNA gene sequences from Lonar lake sediment, India

The data in this article contains the sequences of fungal Internal Transcribed Spacer (ITS) and 18S rRNA gene from a metagenome of Lonar soda lake, India. Sequences were amplified using fungal specific primers, which amplified the amplicon lined between the 18S and 28S rRNA genes. Data were obtained using Fungal tag-encoded FLX amplicon pyrosequencing (fTEFAP) technique and used to analyze fungal profile by the culture-independent method. Primary analysis using PlutoF 454 pipeline suggests the Lonar lake mycobiome contained the 29 different fungal species. The raw sequencing data used to perform this analysis along with FASTQ file are located in the NCBI Sequence Read Archive (SRA) under accession No. SRX889598 (http://www.ncbi.nlm.nih.gov/sra/SRX889598)

Cultivation-independent comprehensive survey of bacterial diversity in Tulsi Shyam Hot Springs, India

A taxonomic description of bacteria was deduced from 5.78 Mb metagenomic sequence retrieved from Tulsi Shyam hot spring, India using bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP). Metagenome contained 10,893 16S rDNA sequences that were analyzed by MG-RAST server to generate the comprehensive profile of bacteria. Metagenomic data are available at EBI under EBI Metagenomics database with accession no. ERP009559. Metagenome sequences represented the 98.2% bacteria origin, 1.5% of eukaryotic and 0.3% were unidentified. A total of 16 bacterial phyla demonstrating 97 families and 287 species were revealed in the hot spring metagenome. Most abundant phyla were Firmicutes (65.38%), Proteobacteria (21.21%) and unclassified bacteria (10.69%). Whereas, Peptostreptococcaceae (37.33%), Clostridiaceae (23.36%), and Enterobacteriaceae (16.37%) were highest reported families in metagenome. Ubiquitous species were Clostridium bifermentans (17.47%), Clostridium lituseburense (13.93%) and uncultured bacterium (10.15%). Our data provide new information on hot spring bacteria and shed light on their abundance, diversity, distribution and coexisting organisms

Isolation, Characterization and investing the Industrial Applications of Thermostable and Solvent Tolerant Serine Protease from Hot Spring Isolated Thermophililic Bacillus licheniformis U1

Protease is the largest selling enzyme in the world due to its various applications in the making of detergent, food and leather, meat tenderisation and pharmaceutical industries. The aim of the study is to isolate and identify thermophilic Bacillus licheniformis U1 strains for thermostable protease production. The partial purified enzyme was characterized under different conditions using Anson-Hagihara’s method. Casein as a substrate in the concentration of 0.6 % w/v optimum for enzyme activity and tolerant up to 2.0% casein concentration. An optimum enzyme activity was reported at pH 7 and decreased with increasing in pH, while temperature optimum was found at 50 °C. The enzyme was stable at 40 °C to 50 °C for half an hour and nearly 50% residual activity was indicated at 60 °C. NaCl was not required for catalysis.  Stability of enzymes in the presence of various organic solvents and different detergents was remarkable. The enzyme was stable up to 3 days into various solvents and slowly denatured with prolonged incubation. The result of the washing performance with detergent was clearly indicated. Moreover the removal of blood stains and dehairing in goat skin suggests the crucial application in the commercial production at large scale.DOI: http://dx.doi.org/10.3126/ijasbt.v2i1.9519Int J Appl Sci Biotechnol, Vol. 2(1): 75-82</jats:p

Contour plots show the response surface effect of interaction on the growth of isolates.

<p>(A) X₁ with X₂, and (B) X₁ with X₄.</p

Zone of syringaldazine oxidation by laccase.

Zone of syringaldazine oxidation by laccase.</p

Enzyme-mediated formulation of stable elliptical silver nanoparticles tested against clinical pathogens and MDR bacteria and development of antimicrobial surgical thread

Abstract Background Silver nanoparticles (AgNPs) are believed to be emerging tool against various infectious diseases including multi-drug resistant (MDR) bacteria. In the present study, in vitro synthesis of AgNPs was optimized using 1:50 ratio of macerozyme (25 μg/μl) and 1 mM AgNO3 incubated at 80 °C for 8 h. AgNPs were characterized by UV–Visible spectroscopy, dynamic light scattering (DLS), scanning electron microscopy, energy-dispersive X-ray spectroscopy, transmission electron microscopy (TEM) and X-ray diffraction (XRD). Results Characterization studies suggest the synthesis of elliptical, stable and crystalline AgNPs with an average size of 38.26 ± 0.4 nm calculated using TEM. The XRD pattern revealed the face-centered-cubic (fcc) form of metallic silver. Good shape integrity and dispersion of AgNPs after 1 year of incubation confirmed their stability. AgNPs were exibited the antimicrobial property against ten pathogenic bacteria, three molds and one yeast. The AgNPs also revealed remarkable antimicrobial activity against three MDR strains i.e. Extended spectrum beta-lactamase positive Escherichia coli, Staphylococcus aureus (MRSA) and Teicoplanin resistant Streptococcus Pneumoniae. The AgNPs coated surgical threads (suture) were revealed the remarkble antibacterial activity against three MDR strains. This is the first report to synthesize antimicrobial elliptical AgNPs using enzymes. Conclusion The results suggest the possibilities to develop the nanoparticles coated antimicrobial medical fabric to combat against MDR infection