Potent Producers Research Articles

Screening and characterization of thermostable enzyme-producing bacteria from selected hot springs of Ethiopia.

Thermostable microbial enzymes play an important role in industries due to their stability under harsh environmental conditions, including extreme temperatures. Despite their huge application in different industries, however, the thermostable enzymes of thermophilic microorganism origin have not yet been fully explored in Ethiopia. Here, we explored thermophilic bacteria and their enzymes from selected hot spring water and sediment samples. Accordingly, thermophilic bacteria were isolated and screened for the production of extracellular hydrolytic enzymes. Promising numbers of isolates were found as producers of the enzymes. The potent enzyme producers were further identified using matrix-assisted laser desorption/ionization time-of-flight-mass spectrometry analysis and 16S rRNA gene sequencing. The findings revealed the presence of potential hydrolytic enzyme-producing thermophilic bacteria in hot springs of Ethiopia and necessitate further comprehensive study involving other extreme environments. Our findings also revealed the potential of Ethiopian hot springs in the production of thermostable enzymes of significant application in different industries, including food industries.

Interspecies metabolite transfer fuels the methionine metabolism of Fusobacterium nucleatum to stimulate volatile methyl mercaptan production.

The major oral odor compound methyl mercaptan (CH3SH) is strongly associated with halitosis and periodontitis. CH3SH production stems from the metabolism of polymicrobial communities in periodontal pockets and on the tongue dorsum. However, understanding of CH3SH-producing oral bacteria and their interactions is limited. This study aimed to investigate CH3SH production by major oral bacteria and the impact of interspecies interactions on its generation. Using a newly constructed large-volume anaerobic noncontact coculture system, Fusobacterium nucleatum was found to be a potent producer of CH3SH, with that production stimulated by metabolic interactions with Streptococcus gordonii, an early dental plaque colonizer. Furthermore, analysis of extracellular amino acids using an S. gordonii arginine-ornithine antiporter (ArcD) mutant demonstrated that ornithine excreted from S. gordonii is a key contributor to increased CH3SH production by F. nucleatum. Further study with 13C, 15N-methionine, as well as gene expression analysis, revealed that ornithine secreted by S. gordonii increased the demand for methionine through accelerated polyamine synthesis by F. nucleatum, leading to elevated methionine pathway activity and CH3SH production. Collectively, these findings suggest that interaction between S. gordonii and F. nucleatum plays a key role in CH3SH production, providing a new insight into the mechanism of CH3SH generation in oral microbial communities. A better understanding of the underlying interactions among oral bacteria involved in CH3SH generation can lead to the development of more appropriate prophylactic approaches to treat halitosis and periodontitis. An intervention approach like selectively disrupting this interspecies network could also offer a powerful therapeutic strategy.IMPORTANCEHalitosis can have a significant impact on the social life of affected individuals. Among oral odor compounds, CH3SH has a low olfactory threshold and halitosis is a result of its production. Recently, there has been a growing interest in the collective properties of oral polymicrobial communities, regarded as important for the development of oral diseases, which are shaped by physical and metabolic interactions among community participants. However, it has yet to be investigated whether interspecies interactions have an impact on the production of volatile compounds, leading to the development of halitosis. The present findings provide mechanistic insights indicating that ornithine, a metabolite excreted by Streptococcus gordonii, promotes polyamine synthesis by Fusobacterium nucleatum, resulting in a compensatory increase in demand for methionine, which results in elevated methionine pathway activity and CH3SH production. Elucidation of the mechanisms related to CH3SH production is expected to lead to the development of new strategies for managing halitosis.

Plant growth promoting potential of Arthrobacter spp., an early colonizer on the volcanic deposits of Mt. Merapi, Indonesia, as a restorative agent for degraded land

Exploration of novel and effective plant growth-promoting bacteria (PGPB) from unexplored environments is an intriguing option for restoring degraded land in Indonesia caused by mining activity. Due to their ability to develop in the absence of carbon, PGPB derived from volcanic environments may be a sustainable alternative. Previous research on the volcanic deposits of Mt. Merapi, Indonesia and Miyake-Jima, Japan, revealed that Arthrobacter spp. dominated the pioneer bacterial communities. Consequently, this research aimed to investigate the biochemical and physiological characteristics of volcanic Arthrobacter groups and assess their potential as PGPB. Using the conventional API system (20NE, 50CH, Zym), biochemical characteristics were evaluated. In vitro evaluations of plant growth-promoting activities such as siderophore production, phosphate solubilization, growth in media devoid of nitrogen, heavy metal resistance and weathering activity were conducted. The results showed broad substrate utilization and enzyme activity profiles of the Mt. Merapi isolates (BRU 37, BRU I.13, and BRU I.45). During 15 days of incubation on a minimal medium containing potassium feldspar, glucose, and NH4Cl, all of the isolates grew and produced acids. In nitrogen-free media, all of the isolates grew. These isolates showed positive reaction for the phosphate solubilization assay using Pikovskaya plates. After one day of incubation, all isolates produced yellow zones on the blue-green pigmented media (CAS-overlay media) and were therefore classified as potent siderophore producers. The heavy metal resistance assay showed that BRU37 grew in the heavy metal media. These results indicate the potential of selected indigenous Mt. Merapi Arthrobacter to enhance plant growth. Further characterization of plant growth-promoting traits is now in progress.

PRODUCTION OF CELLULASE FROM NATURAL BACTERIAL ISOLATES FROM SOIL SAMPLE OF DISTRICT JAUNPUR, U.P., INDIA

The present study aims at isolating, screening, characterizing and optimizing cellulolytic bacteria isolated from soil samples. Different soil samples were collected from different locations of district Jaunpur, Uttar Pradesh, India. The morphological, biochemical and molecular characterization of the isolates were carried out for their identification. A total number of 23 isolates were screened at preliminary level for cellulolytic activity out of which three potent cellulase producers were selected for further study. On the basis of 16S rDNA sequencing these three isolates were identified as Bacillus velezensis PS-14, Bacillus velezensis PS-09 and Bacillus paramycoides PS-05. The optimal conditions for cellulase enzyme production was also determined and maximum activity was obtained at pH 7 and 40 0 C temperature, while 2% inoculum level and 48 hr incubation period were found to be optimum for cellulase production by all the strains. Glucose and Carboxymethyl cellulose was proved to be the best carbon sources, while ammonium choride and peptone was observed to be the most favourable nitrogen source for all three bacterial isolates.

Purification and characterization of arginine deiminase from Klebsiella pneumoniae

Abstract Background and Objectives: This study was aimed to characterize arginine deiminase (ADI) purified from Klebsiella pneumoniae in vitro. Materials and Methods: Precipitation with 70% saturated ammonium sulphate, ion exchange chromatography with a DEAE-cellulose column, and gel filtration chromatography throughout sepharose-6B were the three steps taken to isolate the arginine-degrading enzyme from a K. pneumoniae clinical isolate, which is a potent anticancer source. Results: After 5.9 folds of purification and 38.7% enzyme recovery, the specific activity of the purified enzyme reached 164.2 U/mg. When biochemical characteristics of the purified enzyme were studied, results showed that the activity was maximum at pH 6 and is most stable in pH ranging from (5–9), the optimum temperature for enzyme activity was observed at 37ºC and reach 11.5 U/mL. In contrast, ethylenediaminetetraacetic acid (EDTA), NaNO3, and ZnSO4 slightly inhibited ADI activity, whereas MnCl2, increased the remaining activity of enzyme to 125%., as well as NaNO3, EDTA, ZnSO4, and FeCl3 were found that they inhibit enzyme activity by 90, 70, 88, and 110, respectively. Conclusion: A locally isolated strain of K. pneumoniae N1 is a useful and potent arginine deiminase producer.

Nanoisland SERS-Substrates for Specific Detection and Quantification of Influenza A Virus.

Surface-enhanced Raman spectroscopy (SERS)-based aptasensors for virus determination have attracted a lot of interest recently. This approach provides both specificity due to an aptamer component and a low limit of detection due to signal enhancement by a SERS substrate. The most successful SERS-based aptasensors have a limit of detection (LoD) of 10-100 viral particles per mL (VP/mL) that is advantageous compared to polymerase chain reactions. These characteristics of the sensors require the use of complex substrates. Previously, we described silver nanoisland SERS-substrate with a reproducible and uniform surface, demonstrating high potency for industrial production and a suboptimal LoD of 4 × 105 VP/mL of influenza A virus. Here we describe a study of the sensor morphology, revealing an unexpected mechanism of signal enhancement through the distortion of the nanoisland layer. A novel modification of the aptasensor was proposed with chromium-enhanced adhesion of silver nanoparticles to the surface as well as elimination of the buffer-dependent distortion-triggering steps. As a result, the LoD of the Influenza A virus was decreased to 190 VP/mL, placing the nanoisland SERS-based aptasensors in the rank of the most powerful sensors for viral detection.

Subsistence Economy of Indus People: Agriculture and Domestication of Animal

The importance of Indus civilization stems largely from the fact that it represents the first of the successful achievements of the civilization beyond the bounds of the lands that constitute the fertile crescent. The potency and vigour inherent in food production and domestication of animals were critical in sustaining the large population implied by urbanization. There is a link between development of food production and domestication and the rise of the city life. These processes led to the urbanization in Indus Valley.

Molecular Analysis of Some Virulence Genes of Escherichia Coli Isolates from Wound and Burn Samples in Kirkuk City

Background: Gram-negative infections caused by non-spore producing E. coli that are facultatively anaerobic have been recognized as serious infections linked to higher mortality and significant costs associated with medical care.
 Aim: The goal of this study was to determine whether or not a number of E. coli strains are capable of producing virulence genes by using a technique called real time PCR.
 Materials and Methods: 155 people with burns and wounds had their cultures obtained, and susceptibility testing using swabs, tissue biopsies, and needle aspirations were performed. When compared to other techniques of specimen collection, swab cultures yield precise results and are a practicable, widely utilized, non-invasive, and cost-effective method. The two most recommended techniques for obtaining specimens are tissue biopsy and needle aspiration. Swab cultures, on the other hand, are appropriate since they are useful, widely applied, non-invasive, and reasonably priced. Samples from patients were collected in the consultation rooms, including swabs from burns and wounds. The sufferers had these taken from them. Before being identified, smear samples were initially cultivated in the lab on several selective media. Transport media-filled sterile collection containers containing these samples were brought to the lab.
 Results: According to the study's findings, E. coli was found in 27.10% of the burns and wounds samples. According to the study's findings, the majority of patients with E. coli infections (71.43 percent) were females, while only 28.57 percent were males. According to the study's findings, 28.57 percent of patients diagnosed with E. coli infections were between the ages of 31 and 40. The investigation also indicated that all of the sputum samples containing E. coli isolates were from patients above the age of 40. Furthermore, 45 percent of urine samples were collected from patients aged 31 to 40, and 42.86 percent of patients had wound infections. When examined using the MTP technique, fifty percent of the E. coli isolates tested positive for being potent biofilm producers, according to the study's findings. The DNA of 12 distinct E. coli bacterial isolates was isolated and purified using a genomic DNA purification kit. The bacteria were cultured on chromogenic agar medium to produce these E. coli isolates. The results were detected using electrophoresis on agarose gel at a concentration of 1.5 percent. The gel was then exposed to ultraviolet light, which revealed the DNA as compact bands. The virulence gene was found in 95.24 percent of the E. coli isolates taken from burn and wound sites, according to the study's findings. The majority of E. coli isolates tested positive for amikacin, imipenem, and gentamycin, according to the study's findings. The majority of E. coli isolates, on the other hand, were resistant to the medications amoxicillin, levofloxacin, cefotaxime, ceftriaxone, pefloxacin, cefepime, and ampicillin.

Selenium nanoparticles coated bacterial polysaccharide with potent antimicrobial and anti-lung cancer activities

Bacterial exopolysaccharides are homopolymeric or heteropolymeric polysaccharides with large molecular weights (10–1000 kDa). Exopolysaccharides' functional uses and potential have revolutionized the industrial and medicinal industries. Hence, the aim of the present study was to optimize the production of bacterial exopolysaccharide and apply it as a capping agent for selenium nanoparticles synthesis. Exopolysaccharide (EPS) producing Lactic acid bacteria (LAB) were isolated from dairy products then biochemically characterized and assessed for their potential antimicrobial effect. The most potent EPS producer was identified as Lactiplantibacillus plantarum strain A2 with accession number OP218384 using 16S rRNA sequencing. Overall, FTIR data of the extracted EPS revealed similarity with amylopectin spectrum. 1H NMR spectrum revealed an α-anomeric configuration of the glycosidic linkage pattern in the polysaccharides while the 13C NMR spectrum can also be separated into two main portions, the anomeric carbons region (δ 98–102 ppm) and the non-anomeric carbons region (δ 60–81 ppm). Antimicrobial activity of the produced EPS showed maximum activity against Staphylococcus aureus, MRSA, Enterobacter aerogenes, Klebsiella pneumoniae and Candida albicans respectively. The EPS capsule layer surrounding the bacterial cells was detected by TEM study. Optimization of EPS production was evaluated using Taguchi design, trial 23 reported the highest biomass yield and EPS output (6.5 and 27.12 g/L respectively) with 2.4 and 3.3 folds increase (from the basal media) respectively. The optimized exopolysaccharide was used as a capping and stabilizing agent for selenium nanoparticles (EPS-SeNPs) synthesis. Zeta potential, size and PDI of the synthesized nanoparticles were − 19.7 mV, 45–65 nm and 0.446 respectively with strong bactericidal and fungicidal effect against the tested pathogens. Complete microbial growth eradication was recorded after 6, 8 and 10 h against Staphylococcus aureus, Candida albicans and Klebsiella pneumoniae respectively. EPS-SeNPs showed a potent antioxidant effect reached 97.4% and anticancer effect against A549 lung cancer cell line (IC50 reached 5.324 µg/mL). EPS-SeNPs inhibited cancerous cell growth at S phase. Moreover, molecular studies revealed the anti-apoptotic activity of Bcl2's was inhibited and Bax was activated. The present investigation successfully synthesized selenium nanoparticles through bacterial EPS with significantly high antimicrobial and anticancer activity.

Evaluation of a novel purified and characterized alkaline protease from the extremophile Exiguobacterium alkaliphilum VLP1 as a detergent additive.

This study focused on the isolation and identification of a novel alkaline protease-producing strain from Lake Van, the largest soda lake on Earth. The objective was to purify, characterize, and investigate the potential application of protease in the detergent industry. Through a combination of classical and molecular methods, the most potent protease producer was identified as Exiguobacterium alkaliphilum VLP1. The purification process, involving ammonium sulfate precipitation, ultrafiltration, and anion exchange chromatography, resulted in a 45-fold purification with a yield of 6.4% and specific activity of 1169 Umg-1 protein. The enzyme exhibited a molecular weight of 69kDa, a Km value of 0.4mm, and a maximal velocity (Vmax ) value of 2000 Umg-1 . The optimum activity was observed at 40°C and potential of hydrogen (pH) 9, while the enzyme also exhibited remarkable stability in the ranges of 30-60°C and pH 9-12. Notably, this study represents the first report of an alkaline protease isolated and characterized from E. alkaliphilum. This study also highlighted the potential of the enzyme as a detergent additive, as it showed compatibility with commercial detergents and effectively removed blood and chocolate stains from fabrics.

Improved production of Bacillus subtilis cholesterol oxidase by optimization of process parameters using response surface methodology

BackgroundCholesterol oxidase has numerous biomedical and industrial applications. In the current study, a new bacterial strain was isolated from sewage and was selected for its high potency for cholesterol degradation (%) and production of high cholesterol oxidase activity (U/OD600). ResultsBased on the sequence of 16S rRNA gene, the bacterium was identified as Bacillus subtilis. The fermentation conditions affecting cholesterol degradation (%) and the activity of cholesterol oxidase (U/OD600) of B. subtilis were optimized through fractional factorial design (FFD) and response surface methodology (RSM). According to this sequential optimization approach, 80.152% cholesterol degradation was achieved by setting the concentrations of cholesterol, inoculum size, and magnesium sulphate at 0.05 g/l, 6%, and 0.05 g/l, respectively. Moreover, 85.461 U of cholesterol oxidase/OD600 were attained by adjusting the fermentation conditions at initial pH, 6; volume of the fermentation medium, 15 ml/flask; and concentration of cholesterol, 0.05 g/l. The optimization process improved cholesterol degradation (%) and the activity of cholesterol oxidase (U/OD600) by 139% and 154%, respectively. No cholesterol was detected in the spectroscopic analysis of the optimized fermented medium via gas chromatography-mass spectroscopy (GC–MS). ConclusionThe current study provides principal information for the development of efficient production of cholesterol oxidase by B. subtilis that could be used in various applications.

Application of statistical methodology for the optimization of l-glutaminase enzyme production from Streptomyces pseudogriseolus ZHG20 under solid-state fermentation

BackgroundActinomycetes are excellent microbial sources for various chemical structures like enzymes, most of which are used in pharmaceutical and industrial products. Actinomycetes are preferred sources of enzymes due to their high ability to produce extracellular enzymes. l-glutaminase has proven its essential role as a pharmaceutical agent in cancer therapy and an economic agent in the food industry. The current study aimed to screen the potent l-glutaminase producer and optimize the production media for maximum enzyme yield using one factor at a time (OFAT) approach and statistical approaches under solid-state fermentation (SSF). ResultsOut of 20 actinomycetes strains isolated from rhizosphere soil, 5 isolates produced extracellular l-glutaminase. One isolate was chosen as the most potent strain, and identified as Streptomyces pseudogriseolus ZHG20 based on 16S rRNA. The production and optimization process were carried out under SSF, after optimization using OFAT method, the enzyme production increased up to 884.61 U/gds. Further, statistical strategy, response surface methodology (RSM), and central composite design (CCD) were employed for the level optimization of significant media component (p < 0.05), i.e., wheat bran, sesame oil cake, and corn steep liquor which are leading to increase 3.21-fold l-glutaminase production as compared to unoptimized media. ConclusionsThe presented investigation reveals the optimization of various physicochemical parameters using OFAT and RSM-CCD. Statistical approaches proved to be an effective method for increasing the yield of extracellular l-glutaminase from S. pseudogriseolus ZHG20 where l-glutaminase activity increased up to 1297.87 U/gds which is 3.21-fold higher than the unoptimized medium using a mixture of two solid substrates (wheat bran and sesame oil cake) incubated at pH 7.0 for 6 days at 33 °C.

Analysis, Optimization and Molecular Characterization of PHB Positive Bacteria Isolated from Agricultural Soil Sample

Polyhydroxybutyrates are the widely studied biopolymer because of its biodegradability and non-toxic effect in the environment. This work focused on the isolation, screening, characterization and optimization of the PHB producing bacteria from agricultural soil. About eighty percentage of the isolated bacteria were identified as PHB producing bacteria after Sudan Black Blue staining. Among them three colonies were randomly selected and their morphologically and biochemically characterized. The biopolymer was extracted using sodium hypochlorite method. Potent PHB producer (WJSP1) (13.5µg/ml) was identified by crotonic acid assay. The functional groups of the produced PHB were identified by Fourier Transform Infra-Red spectroscopy (FTIR). The presence of functional groups like carbon group, C=O ester group, CH group, carbonyl group, carboxyl group which was corelated with the peak values1454.33cm-1, 1724.36 cm-1, 1274.95 cm-1, 1049.28 cm-1, 1280-1053 cm-1 indicates the presence of PHB. The potent producer (WJSP1) was optimized under different conditions to achieve the best condition for the production of PHB. The maximum production was obtained at the temperature - 37˚C, the carbon source - glucose, pH - 7 and at a time period of 72 hours. 16SrRNA sequencing was performed and the organism was identified as Bacillus cereus from the BLST (Basic Local Alignment Search Tool). A phylogenetic tree was constructed and the sequence was submitted in GENBANK (Accession No: OR192860). The isolated bacteria Bacillus cereus is an efficient producer of PHB under laboratory condition in presence of glucose as a chief carbon source. Large scale production of PHB can be done in large scale fermenters in industries. Large scale production of PHB will help the environment to get rid of the petroleum-based bioplastics.

Vitamin D regulates glucose metabolism in zebrafish (Danio rerio) by maintaining intestinal homeostasis

Vitamin D (VD) is a steroid hormone that is widely known to play an important role in maintaining mineral homeostasis, and regulating various physiological functions. Our previous results demonstrated that the interruption of VD metabolism caused hyperglycemia in zebrafish. In the present study we further explored the mechanism that VD regulates glucose metabolism by maintaining intestinal homeostasis in zebrafish. Our results showed that the expression of several peptide hormones including gastric inhibitory peptide, peptide YY, and fibroblast growth factor 19 in the intestine decreased, while the expression of sodium glucose cotransporter-1 and gcg was increased in the intestine of the zebrafish fed with the VD3-deficient diet. Consistently, similar results were obtained in cyp2r1−/− zebrafish, in which endogenous VD metabolism is blocked. Furthermore, the results obtained from germ-free zebrafish exhibited that VD-regulated glucose metabolism was partly dependent on the microbiota in zebrafish. Importantly, the transplantation of gut microbiota collected from cyp2r1−/− zebrafish to germ-free zebrafish led to hyperglycemic symptoms in the fish, which were associated with the altered structure and functions of the microbiota in cyp2r1−/− zebrafish. Interestingly, the treatments with acetate or Cetobacterium somerae, a potent acetate producer, lowered the glucose contents whereas augmented insulin expression in zebrafish larvae. Notably, acetate supplementation alleviated hyperglycemia in cyp2r1−/− zebrafish and other diabetic zebrafish. In conclusion, our study has demonstrated that VD modulates the gut microbiota-SCFAs-gastrointestinal hormone axis, contributing to the maintenance of glucose homeostasis.

Non-canonical two-step biosynthesis of anti-oomycete indole alkaloids in Kickxellales

BackgroundFungi are prolific producers of bioactive small molecules of pharmaceutical or agricultural interest. The secondary metabolism of higher fungi (Dikarya) has been well-investigated which led to > 39,000 described compounds. However, natural product researchers scarcely drew attention to early-diverging fungi (Mucoro- and Zoopagomycota) as they are considered to rarely produce secondary metabolites. Indeed, only 15 compounds have as yet been isolated from the entire phylum of the Zoopagomycota.ResultsHere, we showcase eight species of the order Kickxellales (phylum Zoopagomycota) as potent producers of the indole-3-acetic acid (IAA)-derived compounds lindolins A and B. The compounds are produced both under laboratory conditions and in the natural soil habitat suggesting a specialized ecological function. Indeed, lindolin A is a selective agent against plant-pathogenic oomycetes such as Phytophthora sp. Lindolin biosynthesis was reconstituted in vitro and relies on the activity of two enzymes of dissimilar evolutionary origin: Whilst the IAA–CoA ligase LinA has evolved from fungal 4-coumaryl-CoA synthetases, the subsequently acting IAA-CoA:anthranilate N-indole-3-acetyltransferase LinB is a unique enzyme across all kingdoms of life.ConclusionsThis is the first report on bioactive secondary metabolites in the subphylum Kickxellomycotina and the first evidence for a non-clustered, two-step biosynthetic route of secondary metabolites in early-diverging fungi. Thus, the generally accepted “gene cluster hypothesis” for natural products needs to be reconsidered for early diverging fungi.

Phytochemical and metabolic profiling of the different Podocarpus species in Egypt: Potential antimicrobial and antiproliferative activities

Podocarpus is the most dominant genus of Podocarpaceae, with higher taxonomical proximity to the Taxaceae, having numerous pharmaceutical applications, however, scarce studies dealing with the physiological and metabolic criteria of Podocarpus in Egypt were reported. Thus, the objective of this work was to assess the physiological and metabolical patterns of the different species of Podocarpus; P. gracilior, P. elongates, P. macrophyllus and P. neriifolius. The highest terpenoids contents were reported in P. neriifolius, followed by P. elongatus, and P. macrophyllus. P. gracilior had the highest antioxidants amount, followed by P. macrophyllus, P. neriifolius and P. elongatus. From the GC/MS metabolic profiling, caryophyllene, β-cadinene, β-cuvebene, vitispirane, β-cadinene and amorphene were the most dominant metabolites in P. gracilior. β-Caryophyllene was the common in P. gracilior, P. elongatus, P. macrophyllus and P. neriifolius with an obvious fluctuation. The plant methanolic extracts have an obvious activity against the multidrug resistant bacteria; E. coli, P. aeruginosa, S. pyogenes and S. aureus, and fungi; A. fumigatus, A. flavus, A. niger and C. albicans in a concentration-dependent manner. The highest Taxol yield was assessed in the extracts of P. elongatus (16.4 μg/gdw), followed by P. macrophyllus, and P. neriifolius. The chemical identity of Taxol derived from P. elongatus was resolved by LC/MS, with molecular mass 854.6 m/z, and similar structural fragmentation pattern of the authentic one. The highest antitumor activity of P. elongatus extracted Taxol was assessed towards HCT-116 (30.2 μg/ml), HepG-2 (53.7 μg/ml) and MCF-7 (71.8 μg/ml). The ITS sequence of P. elongatus “as potent Taxol producer” was deposited on Genbank with accession #ON540734.1, that is the first record of Podocarpus species on Genbank.

Urban Sewage Canal sediment in Kolkata Metropolis (India) is a potent producer of greenhouse gases

The large network of sewage canals criss-crossing Kolkata Metropolis carry considerable sewage load of about 4.5 million people. In an incubation study, substantial carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O) production were observed in sediment samples collected from the canal bed at five sites, located at city outskirt, urban slum area, industrial area, residential area and agricultural area. The maximum weekly total production of CO2 and CH4 were 201.6, 137.4 mg kg−1 sediment, respectively, while N2O production was 37.1 μg kg−1 sediment. This translated into maximum annual production of 28609.5, 19498.6 and 5.3 t y−1 CO2, CH4 and N2O, respectively, from the sediment pool lying within 0.30 m depth in entire canal network. The CO2-equivalent (100-year time horizon) production in the sediment of canal network was estimated to be 0.56 Tg y−1. The estimated productions underline the chances of substantial emissions of CO2, CH4 and N2O from such large urban sewage canal networks. Sewage canals as GHG source remains under-represented in greenhouse gas (GHG) inventories due to lack of understanding and data on GHG production in canal sediments and their potential subsequent emissions. This study highlights that sewage canals deserve serious consideration in the preparation of GHG budgets in regions hosting large network of sewage canals.

Characterization of Thermostable Cellulase from Bacillus licheniformis PANG L Isolated from the Himalayan Soil.

This study aimed to isolate, purify, and characterize a potential thermophilic cellulase-producing bacterium from the Himalayan soil. Eleven thermophilic bacteria were isolated, and the strain PANG L was found to be the most potent cellulolytic producer. Morphological, physiological, biochemical, and molecular characterization identified PANG L as Bacillus licheniformis. This is the first study on the isolation of thermostable cellulase-producing Bacillus licheniformis from the Himalayan soil. This bacterium was processed for the production of cellulase enzyme. The optimum conditions for cellulase production were achieved at 45°C after 48 h of incubation at pH 6.5 in media-containing carboxymethyl cellulose (CMC) and yeast extract as carbon and nitrogen sources, respectively, in a thermo-shaker at 100 rpm. The enzyme was partially purified by 80% ammonium sulphate precipitation followed by dialysis, resulting in a 1.52-fold purification. The optimal activity of partially purified cellulase was observed at a temperature of 60°C and pH 5. The cellulase enzyme was stable within the pH ranges of 3-5 and retained 67% of activity even at 55°C. Cellulase activity was found to be enhanced in the presence of metal ions such as Cd2+, Pb2+, and Ba2+. The enzyme showed the highest activity when CMC was used as a substrate, followed by cellobiose. The Km and Vmax values of the enzyme were 1.8 mg/ml and 10.92 μg/ml/min, respectively. The cellulase enzyme obtained from Bacillus licheniformis PANG L had suitable catalytic properties for use in industrial applications.

Isolation and Identification of L-asparaginase Producing Bacteria from Soils of Different Agroclimatic Zones of Jammu (J&amp;K), India

A milestone was set in the field of medicine with the discovery of L-asparaginase present in the serum of guinea pig responsible for action against the acute lymphoblastic leukemia. Since then, the use of L-asparaginase as a potential anti-cancer drug has been a great success in the field of modern oncology. Despite its varied sources, new sources are continuously being explored to obtain a high therapeutic index drug. The study aims to isolate potential L-asparaginase producing microorganisms from environmental niches of Northwestern Himalayas. The climatic conditions of the region and ancient agricultural practices which are still being followed in these areas, add up to the diverse microbial repository and a potential habitat which can be explored to obtain a potent asparaginase producing microbial source. In the study, soil samples from different regions of Jammu were collected aseptically. From the isolation studies, a total of 44 bacterial isolates were obtained. Pure cultures were then screened for asparaginase activity both qualitatively as well as quantitatively using agar plate and nesselerization method. Screening studies resulted in the isolation of a potent L-asparaginase producer isolated from black gram soil sample of Rajouri, Jammu showing an enzyme activity of 9.14 U/ml. Based on biochemical and molecular approaches, the isolate was identified as Enterobacter aesburiae strain R16C1 / No. MT93543. The study resulted in the isolation of a pure bacterial culture from the soils of Northwestern Himalayas possessing anti-cancer asparaginase activity. Further, optimization studies can help in considerably increasing the enzyme activity which can be tested against different human cancer lines for its anti-cancer efficacy.

Thrombolytic and anticoagulant efficiencies of purified fibrinolytic enzyme produced from Cochliobolus hawaiiensis under solid-state fermentation.

Cochliobolus hawaiiensis Alcorn Assiut University Mycological Centre 8606 was chosen from the screened 20 fungal species as the potent producer of fibrinolytic enzyme on skimmed-milk agar plates. The greatest enzyme yield was attained when the submerged fermentation (SmF) conditions were optimized, and it was around (39.7U/mg protein). Moreover, upon optimization of fibrinolytic enzyme production under solid-state fermentation (SSF), the maximum productivity of fibrinolytic enzyme was greatly increased recorded a bout (405U/mg protein) on sugarcane bagasse, incubation period of 5 days, moisture level of 100%, initial pH of salt basal medium 7.8, incubation temperature at 35°C, and supplementation of the salt basal medium with corn steep liquor (80％, v/v). The yield of fibrinolytic enzyme by C. hawaiiensis under SSF was higher than that of SmF with about 10.20-fold. The purification procedures of fibrinolytic enzyme by ammonium sulfate (70%), gel filtration, and ion-exchange columns chromatography caused a great increase in its specific activity to 2581.6U/mg protein with an overall yield of 55.89%, 6.37 purification fold and molecular weight of 35kDa. Maximal activity was recorded at pH 7 and 37°C. Significant pH stability was recorded at pH 6.6-7.2, and thermal stability was recorded at 33-41°C. The enzyme showed the highest affinity toward fibrin, with Vmax of 240U/mL and an apparent Km value of 47.61mmol. Mg2+ and Ca2+ moderately induced fibrinolytic activity, whereas Cu2+ and Zn2+ greatly suppressed the enzyme activity. The produced enzyme is categorized as serine protease and non-metalloprotease. The purified fibrinolytic enzyme showed efficient thrombolytic and antiplatelet aggregation activities by completely prevention and dissolution of the blood clot which confirmed by microscopic examination and amelioration of blood coagulation assays. These findings suggested that the produced fibrinolytic enzyme is a promising agent in management of blood coagulation disorders.