Statistical optimization for lipase production from solid waste of vegetable oil industry

<p>The production of biofuel using thermostable bacterial lipase from hot spring bacteria out of low-cost agricultural residue olive oil cake is reported in the present paper. Using a lipase enzyme from <i>Bacillus licheniformis</i>, a 66.5% yield of methyl esters was obtained. Optimum parameters were determined, with maximum production of lipase at a pH of 8.2, temperature 50.8°C, moisture content of 55.7%, and biosurfactant content of 1.693 mg. The contour plots and 3D surface responses depict the significant interaction of pH and moisture content with biosurfactant during lipase production. Chromatographic analysis of the lipase transesterification product was methyl esters, from kitchen waste oil under optimized conditions, generated methyl palmitate, methyl stearate, methyl oleate, and methyl linoleate.</p

Structural insights of microbial community of Deulajhari (India) hot spring using 16s-rRNA based metagenomic sequencing

Insights about the distribution of the microbial community prove to be the major goal of understanding microbial ecology which remains to be fully deciphered. Hot springs being hub for the thermophilic microbiota attract the attention of the microbiologists. Deulajhari hot spring cluster is located in the Angul district of Odisha. Covered within a wooded area, Deulajhari hot spring is also fed by the plant litter resulting in a relatively high amount of total organic content (TOC). For the first time, Illumina sequencing based biodiversity analysis of microbial composition is studied through amplicon metagenome sequencing of 16s rRNA targeting V3‐V4 region using metagenomic DNA from the hot spring sediment. Over 28 phyla were detected through the amplicon metagenome sequencing of which the most dominating phyla at the existing physiochemical parameters like; temperature 69 °C, pH 8.09, electroconductivity 0.025 dSm−1 and total organic carbon 0.356%, were Proteobacteria (88.12%), Bacteriodetes (10.76%), Firmicutes (0.35%), Spirochetes (0.18%) and chloroflexi (0.11%). Approximately 713 species were observed at the above physiochemical parameters. The analysis of the metagenome provides the quantitative insights into microbial populations based on the sequence data in Deulajhari hot spring. Metagenome sequence is deposited to SRA database which is available at NCBI with accession no. SRX1459736. Keywords: Deulajhari hot spring, Illumina sequencing, 16s rRNA, TO

Profiling of microbial community of Odisha hot spring based on metagenomic sequencing

Deulajhari hot spring has diverse temperature and pH range varying from 43 °C to 65 °C and 7.83 to 8.10 respectively. Dense foliage around Deulajhari hot spring contributes to the high total organic carbon content (TOC). In our experiment we took sediment samples from the two Deulajhari hot springs (S1 and S2) out of the cluster having temperature of 43 °C and 55 °C and pH of 7.83 and 7.14 respectively. Sediment samples were analysed using 16S rRNA of V3‐V4 region by amplicon metagenome sequencing. Over 34 phyla were detected in cluster S1 and 32 phyla in cluster S2 at the existing physiochemical parameters temperature 43 °C, pH 7.83, electroconductivity 0.019 dSm−1, and total organic carbon (TOC) 3.80% for S1 and temperature 55 °C, pH 7.14, electroconductivity 0.019 dSm−1, and total organic carbon (TOC) 0.97% for S2. Existence of a vast number of unresolved sequences 179 out of 292 in S1 and 186 out of 314 in S2 at the genus level emphasizes the significance of our study. Metagenome sequence information for the both clusters S1 and S2 of Deulajhari is available at NCBI, SRA database with accession number SRX1459732 and SRX1459733 respectively.Direct link to the deposited data:www.ncbi.nlm.nih.gov/sra/SRX1459732www.ncbi.nlm.nih.gov/sra/SRX1459733 Keywords: Deulajhari, 16S rRNA, Metagenome, TOC, Illumina platfor

Investigation of bacterial diversity of hot springs of Odisha, India

16S rRNA deep sequencing analysis, targeting V3 region was performed using Illumina bar coded sequencing. Sediment samples from two hot springs (Atri and Taptapani) were collected. Atri and Taptapani metagenomes were classified into 50 and 51 bacterial phyla. Proteobacteria (45.17%) dominated the Taptapani sample metagenome followed by Bacteriodetes (23.43%) and Cyanobacteria (10.48%) while in the Atri sample, Chloroflexi (52.39%), Nitrospirae (10.93%) and Proteobacteria (9.98%) dominated. A large number of sequences remained taxonomically unresolved in both hot springs, indicating the presence of potentially novel microbes in these two unique habitats thus unraveling the importance of the current study. Metagenome sequence information is now available at NCBI, SRA database accession no. SRP057428

Investigation of the microbial community in the Odisha hot spring cluster based on the cultivation independent approach

Deulajhari hot spring is located in the Angul district of Odisha. The significance of this hot spring is the presence of the hot spring cluster adjacent to the cold spring which attracts the attention of microbiologists to understand the role of physio-chemical factors of these springs on bacterial community structure. Next-generation sequencing technology helps us to depict the pioneering microflora of any ecological niche based on metagenomic approach. Our study represents the first Illumina based metagenomic study of Deulajhari hot spring DH1, and DH2 of the cluster with temperature 65 °C to 55 °C respectively establishing a difference of 10 °C. Comprehensive study of microbiota of these two hot springs was done using the metagenomic sequencing of 16S rRNA of V3‐V4 region extracting metagenomic DNA from the two hot spring sediments. Sequencing community DNA reported about 28 phyla in spring DH1 of which the majority were Chloroflexi (22.98%), Proteobacteria (15.51%), Acidobacteria (14.51%), Chlorobi (9.52%), Nitrospirae (8.54%), and Armatimonadetes (7.07%), at the existing physiochemical conditions like; temperature 65 °C, pH 8.06, electro conductivity 0.020 dSm−1, and total organic carbon (TOC) 3.76%. About 40 phyla were detected in cluster DH2 at the existing physiochemical parameters like temperature 55 °C, pH 8.10, electro conductivity 0.019 dSm−1, and total organic carbon (TOC) 0.58% predominated with Chloroflexi (41.98%), Proteobacteria (10.74%), Nitrospirae (10.01%), Chlorobi (8.73%), Acidobacteria (6.73%) and Planctomycetes (3.73%). Approximately 68 class, 107 order, 171 genus and 184 species were reported in cluster DH1 but 102 class, 180 order, 375 genus and 411 species in cluster DH2. The comparative metagenomics study of the Deulajhari hot spring clusters DH1, and DH2 depicts the differential profile of the microbiota.Metagenome sequences of these two hot spring clusters are deposited to the SRA database and are available in NCBI with accession no. SRX1459734 for DH1 and SRX1459735 for DH2. Keywords: Deulajhari, 16S rRNA, Next-generation sequencing, Metagenomic

Genetic Stability of Micropropagated Ginger Derived from Axillary Bud through Cytophotometric and RAPD Analysis

A protocol was developed for the in vitro propagation of ginger (Zingiber officinale) cv. Suprava using dormant axillary buds from unsprouted rhizomes. The dormant axillary buds embedded in the rhizome nodes were induced to sprout when cultured on MS medium supplemented with 6-benzyladenine (BA) alone (1Ð6 mg/l) or with a combination of BA (1Ð 6 mg/l) and indole-3-acetic acid (IAA) (0.5, 1 mg/l). In vitro sprouted buds were transferred to the multiplication medium containing various combinations of auxins and cytokinins. MS basal medium supplemented with BA (1 mg/l), IAA (1 mg/l) and adenine sulfate (100 mg/l) was found optimum for the in vitro multiplication of shoots producing (8.2 ð 0.2) shoots from a single explant within 30 days of culture. The multiplication rate remained unchanged in subsequent subcultures. Rooting of shoots occurred in the same multiplication media. Upon transfer of the in vitro culture to ex vitro in pots, 96% of plants survived and established successfully under natural conditions. Tissue culture-raised plantlets of ginger could be conserved in vitro through subculturing at an interval of 4 months. The genetic stability of micropropagated clones was evaluated at regular intervals of 6 months up to 24 months in culture using cytophotometric estimation of 4C nuclear DNA content and random amplified polymorphic DNA (RAPD) analysis. Cytophotometric analysis revealed a unimodal distribution of the DNA content with a peak corresponding to the 4C value (23.1 pg), and RAPD analysis revealed monomorphic bands showing the absence of polymorphism in all fifty regenerants analyzed, thus confirming the genetic uniformity among in vitro grown somaclones of Z. officinale. This study is of commercial significance as axillary bud explants are available throughout the year for initiating a fresh culture of the elite ginger cv. Suprava to be used as a source of true-to-type disease-free planting material thereby minimizing the adverse effect of repeated subculturing from the same explant source

Genetic diversity study of various β-lactamase-producing multidrug-resistant Escherichia coli isolates from a tertiary care hospital using ERIC-PCR

Background & objectives: The prevalence of multidrug-resistant (MDR) Escherichia coli isolates producing β-lactamase enzyme is a growing problem across the globe. Strain typing is an epidemiologically important tool not only for detecting the cross transmission of nosocomial pathogens but also for determining the source of infection. The present study was conducted to understand the clonal relationship among various β-lactamase-producing MDR E. coli isolates using enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR). Methods: A total of 41 MDR E. coli isolates were randomly collected from various clinical samples and processed. Isolated organisms were tested for antibiotics resistance pattern. Phenotypic detection of metallo β-lactamases (MBL) was carried out by the imipenem-ethylenediaminetetraacetic acid disc diffusion/double-disc synergy test. AmpC enzyme production was tested by a modified three-dimensional extract test. Results: Almost all isolates were found sensitive to colistin. A high percentage of drug resistance was observed in these isolates against ceftazidime (100%), cefotaxime (100%), cefepime (100%), ofloxacin (97.56%), amoxicillin/clavulanic acid (97.56%) and norfloxacin (85.36%). Of the 41 isolates, ESBL producers were found to be predominant, i.e., 22 (53.65%), followed by AmpC (6, 14.63%) and MBL (5, 12.19%). Interpretation & conclusions: At 60 per cent similarity cut-off value, the dendrogram analysis showed that there were a total of 14 unique clusters of ERIC (CL-1 - CL-14) within the 41 E. coli isolates, which revealed the genetic diversity existing between them