scholarly journals The chitinolytic activity of the Curtobacterium sp. isolated from field-grown soybean and analysis of its genome sequence

PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0259465
Author(s):  
Ivica Dimkić ◽  
Vibha Bhardwaj ◽  
Valeria Carpentieri-Pipolo ◽  
Nemanja Kuzmanović ◽  
Giuliano Degrassi
Keyword(s):  
Specific Activity ◽  
Gene Clusters ◽  
Defense Responses ◽  
Chitinase Activity ◽  
Glycosyl Hydrolases ◽  
Phylogenetic Cluster ◽  
Genome Visualization ◽  
Chitinase Production ◽  
Or Gene ◽  
Hydrolysis Of

Curtobacterium sp. GD1 was isolated from leaves of conventionally grown soybean in Brazil. It was noteworthy that among all bacteria previously isolated from the same origin, only Curtobacterium sp. GD1 showed a strong chitinase activity. The enzyme was secreted and its production was induced by the presence of colloidal chitin in the medium. The chitinase was partially purified and characterized: molecular weight was approximately 37 kDa and specific activity 90.8 U/mg. Furthermore, Curtobacterium sp. GD1 genome was sequenced and analyzed. Our isolate formed a phylogenetic cluster with four other Curtobacterium spp. strains, with ANIb/ANIm ≥ 98%, representing a new, still non described Curtobacterium species. The circular genome visualization and comparison of genome sequences of strains forming new cluster indicated that most regions within their genomes were highly conserved. The gene associated with chitinase production was identified and the distribution pattern of glycosyl hydrolases genes was assessed. Also, genes associated with catabolism of structural carbohydrates such as oligosaccharides, mixed polysaccharides, plant and animal polysaccharides, as well as genes or gene clusters associated with resistance to antibiotics, toxic compounds and auxin biosynthesis subsystem products were identified. The abundance of putative glycosyl hydrolases in the genome of Curtobacterium sp. GD1 suggests that it has the tools for the hydrolysis of different polysaccharides. Therefore, Curtobacterium sp. GD1 isolated from soybean might be a bioremediator, biocontrol agent, an elicitor of the plant defense responses or simply degrader.

scholarly journals Optimization of Colloidal Chitin and Inoculum Concentration in Chitinase Production by Streptomyces sp. PB2 Using Response Surface Methodology

2020 ◽  
Vol 147 ◽  
pp. 03011
Author(s):  
Eri Pramesti ◽  
Indun Dewi Puspita
Keyword(s):  
Mathematical Model ◽  
Chitinase Activity ◽  
Colloidal Chitin ◽  
Plate Count ◽  
Inoculum Concentration ◽  
Modification Process ◽  
Total Plate Count ◽  
Chitinase Production ◽  
The Mathematical Model ◽  
Hydrolysis Of

Chitinase is an enzyme capable of catalyzing the hydrolysis of chitin. Bacteria is one of the sources for chitinase and the modification process of production are continously developed. Streptomyces sp. is one of Actinomycetes group that shows high activity in hydrolyzing chitin. This study aimed to find the optimal conditions of Streptomyces sp. PB2 for producing chitinase at various colloidal chitin (0.5%, 1%, 1.5%) and inoculum (0.5%, 1%, 1.5%) concentration in the chitin broth medium using Respons Surface Methodology (RSM). The examined parameters included Total Plate Count (CFU/ml) and chitinase activity (U/ml). Chitinase activity was statistically analyzed by MiniTab 17 to obtain a mathematical model, then were validated. The mathematical model of chitinase activity was Y = -0.000075 + 0.00056 K + 0.00067 I, with the optimum colloidal chitin concentration (K) of 1.5% and inoculum concentration (I) of 1.5%. The highest chitinase activity of 0.0019 U/ml in day-2 fermentation. The validation test showed that the mathematical model had a low accuracy with the SSE value of 0.5306. This study shows that colloidal chitin and inoculum concentration are important factors to be optimized. However, further model is needed to be developed for a better estimation of process.

scholarly journals Chitinolytic and Antagonistic Activity of Streptomyces Isolated from Fadama Soil against Phytopathogenic Fungi

2021 ◽  
Vol 32 (3) ◽  
pp. 25-38
Author(s):  
Aminu Argungu Umar ◽  
Aminu Bandam Hussaini ◽  
Jibril Yahayya ◽  
Ibrahim Sani ◽  
Habiba Aminu
Keyword(s):  
Antifungal Agents ◽  
Biocontrol Agent ◽  
Specific Activity ◽  
Extracellular Enzyme ◽  
Chitinase Activity ◽  
Phytopathogenic Fungi ◽  
Antagonistic Activity ◽  
Different Temperatures ◽  
Chitinase Production

Chitinases which degrade chitin have attracted attention as biological antifungal agents. The purpose of this study is to isolate Streptomyces from Fadama soil and assess its chitinolytic and antagonist potential against phytopathogenic fungi for application as biocontrol agent. Streptomyces were isolated from Fadama soil. The selected isolate CT02 exhibited chitinolytic characteristics. Chitinase production was performed under different temperatures, pH and varying incubation period. The highest chitinase production by CT02 isolate was observed after five days of cultivation. The highest chitinase activity was observed at 35°C and pH 7. The crude extracellular enzyme exhibited a specific activity of 4.20 U/μg whereas partially purified extracellular enzyme exhibited a specific activity of 6.19 U/μg with purification fold of 1.47. The selected isolate CT02 and its extracellular crude chitinase showed in vitro antifungal antagonist potential by inhibiting the growth of Aspergillus niger and Aspergillus oryzae. This indicates that Streptomyces derived chitinases are potential biocontrol agents against phytopathogenic fungi.

On the Preparation of Bovine α-Thrombin

1978 ◽  
Vol 39 (01) ◽  
pp. 193-200 ◽  
Author(s):  
Erwin F Workman ◽  
Roger L Lundblad
Keyword(s):  
Immobilized Enzyme ◽  
Catalytic Properties ◽  
Specific Activity ◽  
Guanidine Hydrochloride ◽  
Low Molecular Weight ◽  
Reversible Process ◽  
Gel Beads ◽  
Tissue Thromboplastin ◽  
C 50 ◽  
Hydrolysis Of

SummaryAn improved method for the preparation of bovine α-thrombin is described. The procedure involves the activation of partially purified prothrombin with tissue thromboplastin followed by chromatography on Sulfopropyl-Sephadex C-50. The purified enzyme is homogeneous on polyacrylamide discontinuous gel electrophoresis and has a specific activity toward fibrinogen of 2,200–2,700 N.I.H. U/mg. Its stability on storage in liquid media is dependent on both ionic strenght and temperature. Increasing ionic strength and decreasing temperature result in optimal stability. The denaturation of α-thrombin by guanidine hydrochloride was found to be a partially reversible process with the renatured species possessing properties similar to “aged” thrombin. In addition, the catalytic properties of a-thrombin covalently attached to agarose gel beads were also examined. The activity of the immobilized enzyme toward fibrinogen was affected to a much greater extent than was the hydrolysis of low molecular weight, synthetic substrates.

scholarly journals Salicylic Acid Biosynthesis and Metabolism: A Divergent Pathway for Plants and Bacteria

Biomolecules ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 705
Author(s):  
Awdhesh Kumar Mishra ◽  
Kwang-Hyun Baek
Keyword(s):  
Salicylic Acid ◽  
Bacterial Species ◽  
Shikimate Pathway ◽  
Gene Clusters ◽  
Defense Responses ◽  
Biosynthetic Gene ◽  
Biosynthetic Enzyme ◽  
Biosynthetic Gene Clusters ◽  
Acid Biosynthesis ◽  
Common Precursor

Salicylic acid (SA) is an active secondary metabolite that occurs in bacteria, fungi, and plants. SA and its derivatives (collectively called salicylates) are synthesized from chorismate (derived from shikimate pathway). SA is considered an important phytohormone that regulates various aspects of plant growth, environmental stress, and defense responses against pathogens. Besides plants, a large number of bacterial species, such as Pseudomonas, Bacillus, Azospirillum, Salmonella, Achromobacter, Vibrio, Yersinia, and Mycobacteria, have been reported to synthesize salicylates through the NRPS/PKS biosynthetic gene clusters. This bacterial salicylate production is often linked to the biosynthesis of small ferric-ion-chelating molecules, salicyl-derived siderophores (known as catecholate) under iron-limited conditions. Although bacteria possess entirely different biosynthetic pathways from plants, they share one common biosynthetic enzyme, isochorismate synthase, which converts chorismate to isochorismate, a common precursor for synthesizing SA. Additionally, SA in plants and bacteria can undergo several modifications to carry out their specific functions. In this review, we will systematically focus on the plant and bacterial salicylate biosynthesis and its metabolism.

Characterization of a factor that stimulates hydrolysis of GTP bound to ras gene product p21 (GTPase-activating protein) and correlation of its activity to cell density

1988 ◽  
Vol 8 (10) ◽  
pp. 4169-4173
Author(s):  
M Hoshino ◽  
M Kawakita ◽  
S Hattori
Keyword(s):  
Gene Product ◽  
Cell Density ◽  
Cultured Cells ◽  
Specific Activity ◽  
Gtpase Activating Protein ◽  
Ras Gene ◽  
Cell Extracts ◽  
Mouse Tissues ◽  
Almost All ◽  
Hydrolysis Of

The postmicrosomal fraction of the extract from NIH 3T3 and BALB/c 3T3 cells stimulated the hydrolysis of GTP bound to H-ras gene product p21 by severalfold. The stimulation was observed with normal p21 but not with p21 with valine as the 12th residue. This specificity is similar to that of GTPase-activating protein (GAP) for N-ras p21 described by M. Trahey and F. McCormick (Science 238:542-545, 1987). Consistent with this specificity, analysis of p21-bound nucleotides in living cells revealed that almost all normal p21 bound GDP, whereas oncogenic mutant p21s bound both GTP and GDP. Similar activity was also found in various mouse tissues, with brain tissue showing the highest specific activity. When cell extracts were prepared from cultured cells, there was a linear relationship between GAP activity and cell density. These results suggest the factor is involved in the regulation of cell proliferation.

scholarly journals A Monoacylglycerol Lipase from Mycobacterium smegmatis Involved in Bacterial Cell Interaction

2010 ◽  
Vol 192 (18) ◽  
pp. 4776-4785 ◽  
Author(s):  
Rabeb Dhouib ◽  
Françoise Laval ◽  
Frédéric Carrière ◽  
Mamadou Daffé ◽  
Stéphane Canaan
Keyword(s):  
Mycobacterium Smegmatis ◽  
Wild Type Strain ◽  
Specific Activity ◽  
Cell Interaction ◽  
Monoacylglycerol Lipase ◽  
Homologous Proteins ◽  
Dependent Activity ◽  
Hydrolysis Of

ABSTRACT MSMEG_0220 from Mycobacterium smegmatis, the ortholog of the Rv0183 gene from M. tuberculosis, recently identified and characterized as encoding a monoacylglycerol lipase, was cloned and expressed in Escherichia coli. The recombinant protein (rMSMEG_0220), which exhibits 68% amino acid sequence identity with Rv0183, showed the same substrate specificity and similar patterns of pH-dependent activity and stability as the M. tuberculosis enzyme. rMSMEG_0220 was found to hydrolyze long-chain monoacylglycerol with a specific activity of 143 ± 6 U mg−1. Like Rv0183 in M. tuberculosis, MSMEG_0220 was found to be located in the cell wall. To assess the in vivo role of the homologous proteins, an MSMEG_0220 disrupted mutant of M. smegmatis (MsΔ0220) was produced. An intriguing change in the colony morphology and in the cell interaction, which were partly restored in the complemented mutant containing either an active (ComMsΔ0220) or an inactive (ComMsΔ0220S111A) enzyme, was observed. Growth studies performed in media supplemented with monoolein showed that the ability of both MsΔ0220 and ComMsΔ0220S111A to grow in the presence of this lipid was impaired. Moreover, studies of the antimicrobial susceptibility of the MsΔ0220 strain showed that this mutant is more sensitive to rifampin and more resistant to isoniazid than the wild-type strain, pointing to a critical structural role of this enzyme in mycobacterial physiology, in addition to its function in the hydrolysis of exogenous lipids.

scholarly journals OPTIMALISASI PRODUKSI ENZIM KITINASE PADA ISOLAT JAMUR KITINOLITIK DARI SAMPEL TANAH RIZOSFER

2018 ◽  
Vol 3 (01) ◽  
pp. 62-69
Author(s):  
Eka Corneliyawati ◽  
Massora Massora ◽  
Khikmah Khikmah ◽  
As’ad Syamsul Arifin
Keyword(s):  
Biological Control ◽  
Cell Wall ◽  
Rhizosphere Soil ◽  
Plant Root ◽  
Optimal Growth ◽  
Chitinase Activity ◽  
Fungal Species ◽  
Plant Roots ◽  
Chitinase Production

The rhizosphere is the zone of soil surrounding a plant root where plant roots, soil and the soil biota interact with each other. Chitinolytic fungi has been effectively used in biological control agens. The chitinase activity causes lysis of the fungi cell wall pathogen. The aim of the research was to find optimization of activity chitinase enzyme from rhizosphere soil was conducted in vitro. Optimal growth chitinase production for TKR3 fungi isolate were concentration of chitin 0,2% (b/v), pH 5,5, temperature 30ºC, agitation 150 rpm and incubation time at four days. The optimum yield of chitinase production is influenced by fungal species and environmental conditions.

scholarly journals Structural and Catalytic Characterization of TsBGL, a β-Glucosidase From Thermofilum sp. ex4484_79

2021 ◽  
Vol 12 ◽  
Author(s):  
Anke Chen ◽  
Dan Wang ◽  
Rui Ji ◽  
Jixi Li ◽  
Shaohua Gu ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
Specific Activity ◽  
Maximum Activity ◽  
Homologous Proteins ◽  
Glycosidic Bonds ◽  
Catalytic Sites ◽  
Potential Applications ◽  
Beta Glucosidase ◽  
Hydrolysis Of

Beta-glucosidase is an enzyme that catalyzes the hydrolysis of the glycosidic bonds of cellobiose, resulting in the production of glucose, which is an important step for the effective utilization of cellulose. In the present study, a thermostable β-glucosidase was isolated and purified from the Thermoprotei Thermofilum sp. ex4484_79 and subjected to enzymatic and structural characterization. The purified β-glucosidase (TsBGL) exhibited maximum activity at 90°C and pH 5.0 and displayed maximum specific activity of 139.2μmol/min/mgzne against p-nitrophenyl β-D-glucopyranoside (pNPGlc) and 24.3μmol/min/mgzen against cellobiose. Furthermore, TsBGL exhibited a relatively high thermostability, retaining 84 and 47% of its activity after incubation at 85°C for 1.5h and 90°C for 1.5h, respectively. The crystal structure of TsBGL was resolved at a resolution of 2.14Å, which revealed a classical (α/β)8-barrel catalytic domain. A structural comparison of TsBGL with other homologous proteins revealed that its catalytic sites included Glu210 and Glu414. We provide the molecular structure of TsBGL and the possibility of improving its characteristics for potential applications in industries.

Anterior midgut proteinase inhibitor from Glossina morsitans morsitans Westwood (Diptera: Glossinidae) and its effects upon tsetse digestive enzymes

1980 ◽  
Vol 58 (1) ◽  
pp. 79-87 ◽  
Author(s):  
Jon G. Houseman
Keyword(s):  
Proteinase Inhibitor ◽  
Periplaneta Americana ◽  
Specific Activity ◽  
Glossina Morsitans Morsitans ◽  
Tsetse Flies ◽  
Trypsin Activity ◽  
Glossina Morsitans ◽  
Trypsin Hydrolysis ◽  
Hydrolysis Of ◽  
Anterior Midgut

The anterior midgut of Glossina morsitans morsitans Westwood contains a proteinase inhibitor, molecular weight 5000 ± 2000daltons, stable to 1 M HCl, heat, and dialysis, but unstable to 1% trichloroacetic acid. Inhibitor activity is not associated with anticoagulant in the anterior midgut. The specific activity of the proteinase inhibitor is similar in mated and unmated females and greater than in male tsetse flies. Proteinase inhibitor inhibits proteinase VI and trypsin hydrolysis of N-benzoyl-L-arginine ethyl ester (BAEE) and benzoyl-DL-arginine-p-nitroanilide (BAPNA) but has no effect on proteinase VI hydrolysis of haemoglobin. Inhibition of trypsin hydrolysis of haemoglobin is noncompetitive. Proteinase inhibitor levels in the anterior midgut decreased immediately after feeding and then increased, reaching a maximum 60–100 h after ingestion of the bloodmeal. Postteneral flies contained higher levels of proteinase inhibitor than teneral individuals. Trypsin activity in gut homogenates of Phormia regina and Aedes aegypti was inhibited by the tsetse inhibitor. There was no detectable inhibition of bovine or Pterostichus adstrictus trypsin activity. Inhibition of Periplaneta americana trypsin occurred but was less than fly trypsin inhibition. The possible role of the inhibitor in terminating proteinase production is discussed.

scholarly journals Purification and characterization of a heat-stable esterase from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius

1988 ◽  
Vol 250 (2) ◽  
pp. 453-458 ◽  
Author(s):  
H Sobek ◽  
H Görisch
Keyword(s):  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sulfolobus Acidocaldarius ◽  
Sedimentation Equilibrium ◽  
Prolonged Storage ◽  
Heat Stable ◽  
Equilibrium Data ◽  
Poly Ethylene ◽  
Hydrolysis Of

A heat-stable esterase has been purified 1080-fold to electrophoretic homogeneity from Sulfolobus acidocaldarius, a thermoacidophilic archaebacterium; 20% of the starting activity is recovered. The purified enzyme shows a specific activity of 158 units/mg, based on the hydrolysis of p-nitrophenyl acetate. The esterase hydrolyses short-chain p-nitrophenyl esters, aliphatic esters and triacylglycerols. It is strongly inhibited by paraoxon and phenylmethanesulphonyl fluoride, but only weakly by eserine. From sedimentation-equilibrium data and molecular sieving in polyacrylamide gels, the Mr of the esterase is estimated to be 117000-128000. SDS/polyacrylamide-gel electrophoresis reveals a single band of protein, of Mr 32000. The purified esterase crystallizes in the presence of poly(ethylene glycol) in short rods. The enzyme is inactivated only on prolonged storage at temperature above 90 degrees C.

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