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1.
Heliyon ; 9(4): e15256, 2023 Apr.
Article in English | MEDLINE | ID: mdl-37095930

ABSTRACT

Bacteria residing in contaminated waste soil degrade and utilize organic and inorganic material as a source of nutrients as well as reduce environmental contamination through their enzymatic machinery. This enzymatic potential of indigenous bacteria can be exploited at industrial level through detailed screening, characterization, optimization and purification. In present study, diversity and enzymatic potential of indigenous bacteria was investigated through qualitative and quantitative screening methods from unexplored contaminated soil waste sites in Faisalabad. Shannon diversity (H') index revealed that twenty-eight soil samples from four contaminated sites were highly diverse of amylase, protease and lipase producing bacteria. Maximum protease producing bacteria were detected in fruit waste (1.929 × 107), whereas amylase and lipase producing bacteria were found in industrial (1.475 × 107) and (5.38 × 106), in household waste soil samples. Most of the indigenous bacterial isolates showed potential for multiple enzymes. An isolate OC5 exhibited capability for amylase production and optimization at a wider range of cultural conditions; pH (6-8), temperature (25 °C, 37 °C, 45 °C), incubation time (24-72 h), and NaCl concentrations 0.5-13%, using (1%) starch and lactose as substrates. An isolate OC5 was identified by molecular identification and phylogenetic analysis showed 99% sequence similarity with Bacillus spp. ANOVA was used to analyzed all data statistically. This study enhances the importance of initial screening and reporting of industrially potent indigenous bacteria from unexplored contaminated waste soils. In future, indigenous bacteria in contaminated wastes may be good candidates to solve various environmental pollution problems.

2.
Chem Biodivers ; 20(3): e202200755, 2023 Mar.
Article in English | MEDLINE | ID: mdl-36722706

ABSTRACT

Fraxinus xanthoxyloides is a perennial shrub belonging to family Oleaceae, traditionally used for malaria, jaundice, pneumonia, inflammation, and rheumatism. Our study is aimed to assess the total phenolics (TPC), flavonoids (TFC), terpenoids contents (TTC) and antioxidant profiling of F. xanthoxyloides methanol bark extract (FXBM) and its fractions, hexane, chloroform, ethyl acetate and aqueous, along with high-performance liquid chromatography with diode-array detection (HPLC-DAD). Further, the antioxidant and pulmonary protective potential was explored against carbon tetrachloride (CCl4 )-induced CCl4-induced pulmonary tissue damage in rats. The highest TPC, TFC and TTC were found in FXBM (133.29±4.19 mg/g), ethyl acetate fraction (279.55±10.35 mg/g), and chloroform fraction (0.79±0.06 mg/g), respectively. The most potent antioxidant capacity was depicted by FXBM (29.21±2.40 µg/mg) and ethyl acetate fraction (91.16±5.51 µg/mg). The HPLC-DAD analysis revealed the predominance of gallic, chlorogenic, vanillic and ferulic acid in FXBM. The administration of CCl4 induced oxidative stress, suppressed antioxidant enzymes' activities including catalase, peroxidase, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase, and glutathione reductase. Further, it increased thiobarbituric acid reactive substances (TBARS) and H2 O2 levels, induced DNA injuries and reduced the total protein and glutathione content in lung tissues. The treatment of rats with FXBM restored these biochemical parameters to the normal level. Moreover, the histopathological studies of lung tissues demonstrated that FXBM protected rats' lung tissues from oxidative damage restoring normal lung functions. Thus, F. xanthoxyloides bark extract is recommended as adjuvant therapy as protective agent for patients with lung disorders.


Subject(s)
Antioxidants , Fraxinus , Lung Injury , Plant Extracts , Animals , Rats , Antioxidants/chemistry , Chloroform/metabolism , Chloroform/toxicity , Fraxinus/chemistry , Fraxinus/metabolism , Glutathione/metabolism , Lipid Peroxidation , Liver/metabolism , Lung/metabolism , Oxidative Stress , Plant Bark/chemistry , Plant Bark/metabolism , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Rats, Sprague-Dawley , Lung Injury/chemically induced , Lung Injury/drug therapy
3.
Vet Med Sci ; 9(4): 1861-1868, 2023 07.
Article in English | MEDLINE | ID: mdl-33939314

ABSTRACT

The use of traditional medicines has tremendously increased over the past few decades. Approximately 80% of the world's population relies on traditional medicines for their primary healthcare needs because of their cost effectiveness and efficiency with no or minimal side effects. Zootherapy refers to the use of medicines that are prepared or derived from animals or from their products. The current study documented the folk knowledge related to the practice of various animal-derived products and ethnozoological based drugs used as medicines by the residents of the Cholistan desert of Bahawalpur (Pakistan). In this regard 46 knowledgeable and reliable elderly people, hakims and spiritual healers ranging from 35-60 years of age having knowledge related to zootherapy were included in the current study. A field survey from February 2006 to November 2007 was conducted by interviewing the selected respondents through a structured questionnaire. They provided knowledge regarding the use of animals and their derived products in traditional medicine. The zootherapeutic knowledge was based on both domestic animals as well as wild animals. A total of 20 animal species were included in the study, among which nine animals were domestic while 11 were wild animals. Among selected animals, nine were mammals, four birds, four reptiles and three insects. It was reported that camel was the most commonly used (n = 32 respondents) among mammals while Pigeon (n = 39 respondents), Spiny-tailed lizard (n = 41 respondents) and Indian honey bee (n = 27 respondents) among birds, reptiles and insects, respectively, have significant use for the treatment of different diseases. Based on this communication we could recommend that this type of abandoned knowledge should be considered for the management and conservation of faunistic resources. However, the advantageous role of animals and their products was reported but more extensive research is required to explore the bioactive constituents in the raw material of these animals responsible for their beneficial effects.


Subject(s)
Animals, Wild , Medicine, Traditional , Animals , Bees , Pakistan/epidemiology , Animals, Domestic , Surveys and Questionnaires , Mammals
4.
Saudi J Biol Sci ; 30(2): 103518, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36561334

ABSTRACT

Due to the increased production of polyacrylamide microplastics in the environment impacting the adverse effects on aquatic organisms have become a global concern. The present study aimed to evaluate the toxicity of different concentrations (0.018, 0.03 0.09 g/1) of polyacrylamide microplastics on Oreochromis niloticus. Polyacrylamide microplastics were characterized by Fourier transform infrared and Raman spectroscopy. The FTIR technique revealed the spherical morphology and size range of polyacrylamide (0.1-0.4 mm) with 99 % purity. Raman Spectroscopy results showed peaks around (1100 cm-1 and 1650 cm-1) in gills, these peaks confirmed the presence of polyacrylamide microplastics in the gills of Oreochromis niloticus. Polyacrylamide microplastics significantly shortened the antioxidant enzymes (Catalase and Reduced glutathione) proceedings and the increase was observed in Malondialdehyde and Lipid peroxide levels in high-dose treated groups. Moreover, total protein contents were expressively increased, while other blood parameters (AST, ALP, ALT) were significantly decreased. Hemoglobin (g/dl), Erythrocyte (106/µl) and TLC (1x109/l) levels were reduced significantly (p < 0.05) in high concentrations. The administration of polyacrylamide microplastics induced different histological changes in the gills, liver and intestine of O. niloticus. It was concluded that polyacrylamide microplastics are toxic agents having harmful effects on fish health.

5.
Molecules ; 26(19)2021 Sep 30.
Article in English | MEDLINE | ID: mdl-34641480

ABSTRACT

The present research is based on the fabrication preparation of CS/PVA/GG blended hydrogel with nontoxic tetra orthosilicate (TEOS) for sustained paracetamol release. Different TEOS percentages were used because of their nontoxic behavior to study newly designed hydrogels' crosslinking and physicochemical properties. These hydrogels were characterized using Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and wetting to determine the functional, surface morphology, hydrophilic, or hydrophobic properties. The swelling analysis in different media, degradation in PBS, and drug release kinetics were conducted to observe their response against corresponding media. The FTIR analysis confirmed the components added and crosslinking between them, and surface morphology confirmed different surface and wetting behavior due to different crosslinking. In various solvents, including water, buffer, and electrolyte solutions, the swelling behaviour of hydrogel was investigated and observed that TEOS amount caused less hydrogel swelling. In acidic pH, hydrogels swell the most, while they swell the least at pH 7 or higher. These hydrogels are pH-sensitive and appropriate for controlled drug release. These hydrogels demonstrated that, as the ionic concentration was increased, swelling decreased due to decreased osmotic pressure in various electrolyte solutions. The antimicrobial analysis revealed that these hydrogels are highly antibacterial against Gram-positive (Staphylococcus aureus and Bacillus cereus) and Gram negative (Pseudomonas aeruginosa and Escherichia coli) bacterial strains. The drug release mechanism was 98% in phosphate buffer saline (PBS) media at pH 7.4 in 140 min. To analyze drug release behaviour, the drug release kinetics was assessed against different mathematical models (such as zero and first order, Higuchi, Baker-Lonsdale, Hixson, and Peppas). It was found that hydrogel (CPG2) follows the Peppas model with the highest value of regression (R2 = 0.98509). Hence, from the results, these hydrogels could be a potential biomaterial for wound dressing in biomedical applications.


Subject(s)
Anti-Bacterial Agents/administration & dosage , Bacteria/drug effects , Chitosan/chemistry , Galactans/chemistry , Hydrogels/administration & dosage , Mannans/chemistry , Plant Gums/chemistry , Polyvinyl Alcohol/chemistry , Wound Healing/drug effects , Anti-Bacterial Agents/chemistry , Bandages , Drug Liberation , Hydrogels/chemistry
6.
Heliyon ; 7(5): e07030, 2021 May.
Article in English | MEDLINE | ID: mdl-34095563

ABSTRACT

The present study investigated the (i) socio-demographic predictors of psychological distress, (ii) socio-demographic predictors of satisfaction from online classes, and (iii) the relationship between psychological distress and satisfaction from online classes among university students of Pakistan during the COVID-19 pandemic. An online questionnaire-based survey was conducted. A total of 2220 respondents that was enrolled at the University of the Punjab (PU), University of Management and Technology (UMT), and the University of Central Punjab (UCP) were involved in the current study. Data were collected at a 64% response rate and analyzed with SPSS IBM Version 21.0. Results revealed that approximately 41% of the students were facing severe psychological distress while about 65% were found unsatisfied with online classes. Besides, a linear negative relationship between the independent variable, i.e. psychological distress and the dependent variable, i.e. satisfaction from online classes was found. Therefore, to minimize the level of psychological distress and increase students' satisfaction with online classes it is highly recommended to take precautionary measures by the relevant stakeholders.

7.
Saudi J Biol Sci ; 27(12): 3499-3504, 2020 Dec.
Article in English | MEDLINE | ID: mdl-33304161

ABSTRACT

Amylases take part with vital role in industries such as food, fermentation; starch processing, textile and paper etc. Increasing amylases demand, high nutrient expenditure and environmental pollution have forced to utilize agro-industrial residues as a low-cost feedstock for enzyme production. In present study, three soil samples were collected from agro-industrial waste dumping areas in District Faisalabad. Ten thermophilic bacterial isolates were separated at 55 °C on the basis of colonial morphology, three isolates (F6, F11, F17) showed prominent zone of clearance applying iodine test on starch agar plates. Bacterial isolate F-11 showed highest amylase activity with DNS method and molecularly identified through 16S RNA sequencing as Bacillus sp. with Accession number MH917294. Four unconventional food wastes (banana, lemon, mango and potato) pretreated with 0.8% sulphuric acid concentrations taking 1000 g/L weight released the highest sugars contents and phenolic components. Maximum amylase activity i.e. 29.23 mg/ml was achieved in mango waste at, 40 °C, with pH 6.0 and 0.17% nitrogenous source adding 8% inoculum size (2 days old) using Response Surface Methodology (RSM) for optimization. Crude amylase confirmed its efficiency in starch hydrolysis that suggested it as potential candidate for application in starch industries.

8.
Acta Microbiol Immunol Hung ; 67(2): 87-90, 2020 Jul 03.
Article in English | MEDLINE | ID: mdl-32619190

ABSTRACT

World has been suffering from pandemic caused by mysterious Coronavirus. The novel member of Coronaviridae causing COVID-19 disease is named as SARS-Cov-2. Its first case was reported in China by the end of 2019, but its exponential spread has wrapped entire globe, suspended and is penalizing mankind. A retrospective meta-analysis study showed that outbreaks of Middle East Respiratory Syndrome (MERS) and SARS-Cov-1 (Coronaviridae), influenza infection H1N1 and West-African Ebola caused lower mortality than this new pandemic COVID-19. Virus has appeared as a new human pathogen so to counter COVID-19 no specific vaccine, monoclonal antibodies have been manufactured till day. The outbreak of novel Coronavirus is treated with antimicrobial drugs but they have their own mild side effects. But the drastic spread of COVID-19 compels us also to use other ways to counter pandemic. Thus, passive immunization can be opted to hold back this mysterious virus. Passive immunization has been in use since early 20th century and showed its effectiveness against all previous infectious outbreaks including MERS and SARS-Cov-1 members of Coronaviridae. The review argues that convalescent plasma is an explicit option for containment of COVID-19 disease.


Subject(s)
Antibodies, Viral/therapeutic use , COVID-19/therapy , Immunization, Passive/methods , Adult , Aged , COVID-19/mortality , COVID-19/pathology , Female , Humans , Immunotherapy/methods , Male , Middle Aged , Retrospective Studies , SARS-CoV-2/drug effects , SARS-CoV-2/immunology , COVID-19 Serotherapy
9.
RSC Adv ; 10(17): 10221-10231, 2020 Mar 06.
Article in English | MEDLINE | ID: mdl-35498604

ABSTRACT

Secondary metabolites present in medicinal plants offer a golden opportunity to fight different ailments, such as cancer, infections, diabetes, neurodegenerative and cardiovascular diseases, etc. The traditional use of various parts of Fraxinus xanthoxyloides is known to serve as a cure for pneumonia, pain, jaundice, malaria, fracturing of bones, and internal wounds. The aim of this research was to validate the antioxidant and cardio-protective properties of F. xanthoxyloides leaves. The antioxidant potential was evaluated by employing different assays on the crude methanol extract, as well as its derived fractions. The extract/fraction that showed significant activity was further investigated for the presence of phytochemicals using high performance liquid chromatography-diode array detector (HPLC-DAD) analysis and also for cardio-protective potential. In the case of the antioxidant potential, the ethyl acetate fraction (FXE) was demonstrated to have the most potent total antioxidant (26.3 ± 2.4 AAE µg mg-1), hydroxyl ion scavenging (IC50 = 7.9 ± 0.9 µg mg-1), ferrous ion chelating (IC50 = 28.2 ± 2.7 µg mg-1) and nitric oxide scavenging (IC50 = 32.5 ± 2.9 µg mg-1) effects among all of the extract/fractions, whereas in the case of DPPH (IC50 = 17.5 ± 2.7 µg mg-1) and the reducing power assay (16.7 ± 2.8 GAE µg mg-1), promising antioxidant potential was shown by the n-butanol fraction. The presence of different concentrations of rutin, caffeic acid, catechin, and gallic acid was observed in the high performance liquid chromatography (HPLC) profile of FXE. Furthermore, in in vivo experimentation, the oral administration of FXE and silymarin significantly restored the CCl4-induced increase in the levels of creatine kinase, creatine kinase-MB, cholesterol and triacylglycerides when compared with the untreated group. FXE and silymarin treatment also restored the levels of the tissue antioxidant enzymes, for example glutathione-S-transferase, glutathione reductase, catalase, peroxidase and superoxide dismutase. Furthermore, significantly lower levels of reduced glutathione and enhanced levels of lipid peroxides, hydrogen peroxide, comet length and DNA damages were observed after CCl4 administration in the cardiac tissue of rats. FXE was able to restore these biochemical parameters, as well as the histological status of heart tissue. Based upon the present investigation, we concluded that F. xanthoxyloides leaves may have cardio-protective potential similar to silymarin against CCl4 induced injuries owing to its antioxidant constituents.

10.
Pak J Pharm Sci ; 32(5(Supplementary)): 2237-2243, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31894049

ABSTRACT

Hispolon, a bioactive polyphenolic entity extracted from Phellinus linteus, possesses anticancer, antiinflammatory and anti-oxidant properties. Despite the reported therapeutic effects of this natural chemical entity, inhibitory potential of hispolon towards prostate carcinoma DU145 cells and mechanism of its action are yet to be explicated. Deregulated STAT3 pathway performs multifaceted functions in facilitating the development of cancer. Here, we have investigated the mechanism of hispolon by which it exerts its anticancer effects in DU145 cells and whether its anticancer activity is mediated by modulation of STAT3. Our outcomes show that hispolon significantly halted the multiplication of DU145 cells as well as arrested cell cycle at S phase. S phase arrest induced by hispolon was associated with downregulation of cyclin B1, cyclin D1 and CDK4 while up-regulation of p21. Moreover, hispolon treatment leads towards induction of apoptosis in a dose-dependent mode in DU145 cells. Hispolon induced modulation of Bcl-2 family proteins lead towards loss of MMP allowing the discharge of cytochrome c from mitochondrial porin channels which triggered the cascade of caspases ultimately causing cellular death. We further investigated the role of hispolon in mediating deregulated STAT3 pathways in DU145 cells. Hispolon has potential to downregulate the p-STAT3 expression with no effect on total STAT3. Contemporaneously, these results represent that hispolon's anticancer mechanism of action proceeds via downregulating the phosphorylation of STAT3 and induction of apoptosis via mitochondrial pathway.


Subject(s)
Apoptosis/drug effects , Catechols/pharmacology , Mitochondria/drug effects , Prostatic Neoplasms/drug therapy , STAT3 Transcription Factor/physiology , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , Flow Cytometry , Humans , Male , Phosphorylation , Prostatic Neoplasms/pathology , Proto-Oncogene Proteins c-bcl-2/analysis , Signal Transduction/drug effects
11.
Pak J Pharm Sci ; 31(4(Supplementary)): 1475-1484, 2018 Jul.
Article in English | MEDLINE | ID: mdl-30058538

ABSTRACT

Roots, bark, stem/twigs, and leaves of Fraxinus xanthoxyloides are being used regionally for the cure of malaria, jaundice, internal injuries, pneumonia, pain, rheumatism and also in fracture of bones. Our objective was to assess the methanolic leaves extract of F. xanthoxyloides for its antioxidant capability against oxidative stress induced by carbon tetrachloride (CCl4) in the kidney of Sprague-Dawley rats. Duration of this experiment was 30 days and doses were given on alternative days. Urine of rats was assessed for kidney function and renal tissues for antioxidant enzymes activity, biochemical markers, comet assay and histopathology. Enhanced urinary creatinine, urobilinogen levels and decreased creatinine clearance, protein contents, and albumin levels were observed by CCl4 administration when matched to controls. CCl4 injection also decreased the level of reduced glutathione, catalase, super oxide dismutase, peroxidase, glutathione s-transferase, glutathione reductase, and tissue protein while elevated the levels of thiobarbituric acid reactive substances, DNA damages and H2O2 in renal tissues of experimental animals. Co-treatment of FXM and silymarin, lead to the restoration of all the above tested parameters of kidney. Through this study we affirmed the ameliorating role of F. xanthoxyloides in oxidative stress affiliated disorders of kidney.


Subject(s)
Acute Kidney Injury/chemically induced , Acute Kidney Injury/drug therapy , Carbon Tetrachloride/toxicity , Fraxinus , Plant Extracts/therapeutic use , Plant Leaves , Acute Kidney Injury/pathology , Animals , Male , Methanol/therapeutic use , Plant Extracts/isolation & purification , Rats , Rats, Sprague-Dawley
12.
J Photochem Photobiol B ; 179: 126-133, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29367147

ABSTRACT

Biodiesel is produced by transesterification of fatty acid methyl esters (FAME) from oleaginous microalgae feedstock. Biodiesel fuel properties were studied and compared with biodiesel standards. Qualitative analysis of FAME was done while cultivating mixed microalgae consortia under three concentrations of coal fired flue gas (1%, 3.0% and 5.5% CO2). Under 1% CO2 concentration (flue gas), the FAME content was 280.3 µg/mL, whereas the lipid content was 14.03 µg/mL/D (day). Both FAMEs and lipid contents were low at other CO2 concentrations (3.0 and 5.5%). However, mixed consortia in the presence of phosphate buffer and flue gas (PB + FG) showed higher saturated fatty acids (SFA) (36.28%) and unsaturated fatty acids (UFA) (63.72%) versus 5.5% CO2 concentration, which might be responsible for oxidative stability of biodiesel. Subsequently, higher cetane number (52) and low iodine value (136.3 gI2/100 g) biodiesel produced from mixed consortia (PB + FG) under 5.5% CO2 along with 50 mM phosphate buffer were found in accordance with European (EN 14214) standard. Results revealed that phosphate buffer significantly enhanced the biodiesel quality, but reduced the FAME yield. This study intended to develop an integrated approach for significant improvement in biodiesel quality under surplus phosphorus by utilizing waste flue gas (as CO2 source) using microalgae. The CO2 sequestration from industrial flue gas not only reduced greenhouse gases, but may also ensure the sustainable and eco-benign production of biodiesel.


Subject(s)
Biofuels , Carbon Dioxide/metabolism , Fatty Acids/analysis , Microalgae/metabolism , Biomass , Carbon Dioxide/chemistry , Chromatography, Gas , Coal/analysis , Fatty Acids/metabolism , Gases/chemistry , Mass Spectrometry , Microalgae/growth & development
13.
Int Microbiol ; 21(4): 163-174, 2018 Dec.
Article in English | MEDLINE | ID: mdl-30810899

ABSTRACT

Massive production of keratinaceous byproducts in the form of agricultural and industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This review summarizes the potential utility of some bacterial and fungal species for the production of keratinase using a variety of keratinaceous wastes as growth substrates. The application of microbial keratinases in waste management; animal feed, detergent, and fertilizer manufacturing; and leather, cosmetic, and pharmaceutical industries is also abridged in this review.


Subject(s)
Bacteria/enzymology , Fungi/enzymology , Industrial Microbiology/methods , Keratins/metabolism , Peptide Hydrolases/metabolism , Agriculture/methods , Bacteria/metabolism , Biotechnology/methods , Biotransformation , Fungi/metabolism , Industrial Waste , Waste Management/methods
14.
Carbohydr Polym ; 136: 551-9, 2016 Jan 20.
Article in English | MEDLINE | ID: mdl-26572387

ABSTRACT

Thermally-induced phase separation (TIPS) method was used to synthesize polymer matrix (PM) membranes for reverse osmosis from cellulose acetate/polyethylene glycol (CA/PEG300) conjugated with silica nanoparticles (SNPs). Experimental data showed that the conjugation of SNPs changed the surface properties as dense and asymmetric composite structure. The results were explicitly determined by the permeability flux and salt rejection efficiency of the PM-SNPs membranes. The effect of SNPs conjugation on MgSO4 salt rejection was more significant in magnitude than on permeation flux i.e. 2.38 L/m(2)h. FTIR verified that SNPs were successfully conjugated on the surface of PM membrane. DSC of PM-SNPs shows an improved Tg from 76.2 to 101.8 °C for PM and PM-S4 respectively. Thermal stability of the PM-SNPs membranes was observed by TGA which was significantly enhanced with the conjugation of SNPs. The micrographs of SEM and AFM showed the morphological changes and increase in the valley and ridges on membrane surface. Experimental data showed that the PM-S4 (0.4 wt% SNPs) membrane has maximum salt rejection capacity and was selected as an optimal membrane.


Subject(s)
Cellulose/analogs & derivatives , Membranes, Artificial , Nanoparticles/chemistry , Osmosis , Polyethylene Glycols/chemistry , Silicon Dioxide/chemistry , Cellulose/chemistry , Magnesium Sulfate/chemistry
15.
Nat Prod Res ; 30(6): 720-3, 2016.
Article in English | MEDLINE | ID: mdl-26373305

ABSTRACT

In this work chitin was extracted chemically from shrimp shells. Seventeen Bacillus isolates were screened for chitinolytic activity. The chitinolytic strains of Bt. were screened at different temperatures and pHs for their hydrolytic potentials. By using a pair of specific primers, endochitinase gene was amplified from SBS Bt-5 strain through PCR, and then cloned into pTZ57 TA cloning vector and transferred in Escherichia coli DH5α strain. The sequenced gene (GenBank Accession No: HE995800) consists of 2031 nucleotides capable of encoding 676 residues. The protein consisted of three functional domains with a calculated molecular mass of 74.53 kDa and a pI value of 5.83. The amino acid sequence of chi gene showed 99% similarity to the genes of Bt MR11 endochitinase, Bt serovar kurstaki chitinase (kchi), Bt strain MR21 endochitinase and Bacillus cereus B4264.


Subject(s)
Bacillus/enzymology , Bacterial Proteins/genetics , Chitin/metabolism , Chitinases/genetics , Amino Acid Sequence , Bacillus/genetics , Cloning, Molecular , DNA, Bacterial/genetics , Escherichia coli , Polymerase Chain Reaction , Sequence Analysis, DNA
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