Expression, purification, crystallization and preliminary X-ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum

Conjugated bile salt hydrolase (BSH) catalyses the hydrolysis of the amide bond that conjugates bile acids to glycine and to taurine. The BSH enzyme from Bifidobacterium longum was overexpressed in Escherichia coli BL21(DE3), purified and crystallized. Crystallization conditions were screened using the hanging-drop vapour-diffusion method. Crystal growth, with two distinct morphologies, was optimal in experiments carried out at 303 K. The crystals belong to the hexagonal system, space group P622 with unit-cell parameters a = b = 124.86, c = 219.03 Angstrom, and the trigonal space group P321, with unit-cell parameters a = b = 125.24, c = 117.03 Angstrom. The crystals diffracted X-rays to 2.5 Angstrom spacing. Structure determination using the multiple isomorphous replacement method is in progress

Expression, purification, crystallization and preliminary X-ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum

Conjugated bile salt hydrolase (BSH) catalyses the hydrolysis of the amide bond that conjugates bile acids to glycine and to taurine. The BSH enzyme from Bifidobacterium longum was overexpressed in Escherichia coli BL21(DE3), purified and crystallized. Crystallization conditions were screened using the hanging-drop vapour-diffusion method. Crystal growth, with two distinct morphologies, was optimal in experiments carried out at 303 K. The crystals belong to the hexagonal system, space group P622 with unit-cell parameters a = b = 124.86, c = 219.03 Angstrom, and the trigonal space group P321, with unit-cell parameters a = b = 125.24, c = 117.03 Angstrom. The crystals diffracted X-rays to 2.5 Angstrom spacing. Structure determination using the multiple isomorphous replacement method is in progress

X-ray studies on crystalline complexes involving amino acids and peptides. Part XIV. Closed conformation and head-to-tail arrangement in a new crystal form of L-histidine L-aspartate monohydrate

A new form of L-histidine L-aspartate monohydrate crystallizes in space group P21 with a = 5·131(1), b = 6·881(1), c= 18·277(2) Å, β= 97·26(1)° and Z = 2. The structure has been solved by the direct methods and refined to an R value of 0.044 for 1377 observed reflections. Both the amino acid molecules in the complex assume the energetically least favourable allowed conformation with the side chains staggered between the α-amino and α-carboxylate groups. This results in characteristic distortions in some bond angles. The unlike molecules aggregate into alternating double layers with water molecules sandwiched between the two layers in the aspartate double layer. The molecules in each layer are arranged in a head-to-tail fashion. The aggregation pattern in the complex is fundamentally similar to that in other binary complexes involving commonly occurring L amino acids, although the molecules aggregate into single layers in them. The distribution of crystallographic (and local) symmetry elements in the old form of the complex is very different from that in the new form. So is the conformation of half the histidine molecules. Yet, the basic features of molecular aggregation, particularly the nature and the orientation of head-to-tail sequences, remain the same in both the forms. This supports the thesis that the characteristic aggregation patterns observed in crystal structures represent an intrinsic property of amino acid aggregation

Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants

The crystallization of three catalytically inactive mutants of penicillin Vacylase (PVA) from Bacillus sphaericus in precursor and processed forms is reported. The mutant proteins crystallize in different primitive monoclinic space groups that are distinct from the crystal forms for the native enzyme. Directed mutants and clone constructs were designed to study the post-translational autoproteolytic processing of PVA. The catalytically inactive mutants will provide threedimensional structures of precursor PVA forms, plus open a route to the study of enzyme-substrate complexes for this industrially important enzyme

Analysing Amino Acids in Galanin Graph Theoretical Approach

Graph theoretical analysis is an important area of the research in biological networks. Here first we introduce Pt-graph of peptide/protein based on physicochemical properties and adjacency of amino acids in the corresponding peptide/protein. Based on the Pt-graph, we introduce the graph of species which containing the peptide/protein named as SPt-graph. Finally, we analyze graph theoretically Pt-graphs of fourteen species of animals containing Galanin, a neuropeptide and their SPt-graph. From the graph theoretical analyses of Pt-graph and SPt-graph we get some observations about the relations among the amino acids, physicochemical properties of amino acids, peptide/protein and species containing peptide/protein and it may help in the field of evolution of peptide/protein and drug design in future

A STUDY ON MODERN TECHNIQUES USED IN IRRIGATION FOR FARMING IN COIMBATORE CITY

Water is most powerful natural resource it is mostly influences in health and wealth of the people and production of food. India economic backbone is agricultural. We are depending the water resources for development agriculture. Irrigation plays main role in the food production. The future expansion of food production will be increasing dependents upon sound irrigation and water management systems and now days the resources are most challenging because of environment changes. These days’ people prefer to use modern irrigational techniques because they are extremely efficient and effective. Modern irrigational techniques are also pivotal in increasing the crop yield. Also since these days farming is done with the help of fertilizers, crops need a certain amount of water to grow properly. Modern irrigational techniques are also cost effective, if done on a large scale. Also modern irrigational methods are important because of scarcity of water and increasing demand for food crops. For example: drip irrigation, if done properly can save up to 95% of water. According to me, modern irrigational techniques should be used and promoted because they save a lot of water

A meglumine catalyst–based synthesis, molecular docking, and antioxidant studies of dihydropyrano[3, 2-b]chromenedione derivatives

A simple method was employed for the synthesis of dihydropyrano[3, 2-b]chromenedione derivatives (4a-o) in high yields by condensation of 5, 5-dimethylcyclohexane-1, 3-dione(1), different aromatic aldehydes (2a-o), and 5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one(3), using meglumine as a stable and reusable catalyst. Meglumine, an amino sugar, was employed as an environmentally benign catalyst, due to its splendid properties such as being inexpensive, recyclable, and biodegradable. The accomplished protocol employs low catalyst loading and easy work-up for the synthesis of 5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one derivatives. A great asset is that without any significant loss, the catalyst could be recovered and reused for extended synthetic steps. This offer huge advantage to overcome recyclability issues. Our synthesized compounds were analyzed by IR, 1H, 13C NMR, mass spectra and evaluated for their antioxidant properties by 1, 1-diphenyl-2-picryl hydrazyl radical (DPPH), hydrogen peroxide(H2O2), and nitric oxide (NO) scavenging methods. The correlation in exhibition of antioxidant activity was effective at all doses. The binding interactions and molecular docking studies for entitled compounds were studied against 3MNG protein; 4k exhibited marked binding affinity with excellent docking score of −7.6 Kcal/mol and emerged as a lead compound. © 2019 Wiley Periodicals, Inc.Ural Federal University, UrFUThe authors G. Sravya and N. Bakthavatchala Reddy are thankful to Ural Federal University, Yekaterinburg, Russia, for postdoctoral fellowship

Magnetic and the magnetocaloric properties of Ce1-xRxFe2 and Ce(Fe1-xMx)2 compounds

We have studied selected rare earth doped and transition metal doped CeFe2 compounds by examining their structural, magnetic and magneto-thermal properties. With substitution of Ce by 5 and 10% Gd and 10% Ho, the Curie temperature can be tuned to the range of 267-318 K. Localization of Ce 4f electronic state with rare earth substitutions is attributed for the enhancement of Curie temperature. On the other hand, with Ga and Al substitution at the Fe site, system undergoes paramagnetic to ferromagnetic transition and then to an antiferromagnetic phase on cooling. The magnetocaloric effect across the transitions has been studied from both magnetization isotherms and heat capacity data. It is shown that by choosing the appropriate dopant and its concentration, the magnetocaloric effect around room temperature can be tuned.Comment: 13 pages, 6 figures, 2 table

First and second trimester bleeding and pregnancy outcome: a prospective study in a tertiary government hospital

Background: Bleeding in first and second trimester of pregnancy is one of the common complications of pregnancy. there is evidence from various prospective and retrospective studies that first and second trimester vaginal bleeding which continue with pregnancy is associated with adverse pregnancy outcome, including preterm delivery, low birth weight babies, perinatal death and congenital anomalies. Objective of this study was to know the outcome of pregnancies who have bleeding in first and second trimester of pregnancy.Methods: This study was prospective study done in the department of obstetrics and gynaecology, Vanivilas Hospital, Bangalore from September 2018 to August 2019.Results: This study concludes that I trimester vaginal bleeding are at increased risk of abortion than in II trimester vaginal bleeding. Risk of placenta previa was more in II trimester vaginal bleeding than in I trimester vaginal bleeding.Conclusions: This study concludes that I trimester vaginal bleeding are at increased risk of abortion than in II trimester vaginal bleeding. Risk of placenta previa was more in II trimester vaginal bleeding than in I trimester vaginal bleeding. Bleeding in I trimester and II trimester call for special attention in view of increased risk of preterm birth and perinatal death. Recognition of these association will be useful for detection and follow up of pregnancies being at high risk

A prospective study of role of doppler in pregnancy and the perinatal outcome

Background: The development of doppler ultrasonographic technology has provided an opportunity to obtain a qualitative and quantitative assessment of maternal and foetal circulation using a non-invasive method. It has been proved by many studies that doppler has a very important role in screening of high-risk pregnancies. Objective of this study was to evaluate the role of colour doppler study in normal and high-risk pregnancy in relation to perinatal outcome.Methods: A prospective study was done including 75 women with high risk pregnancy and 75 normal pregnant women during the period October 2018 to September 2019 in hospitals attached to Bangalore Medical College and Research Institute. Doppler examination was done after recording patients’ history, clinical examination and ultrasound. Results were analysed and conclusions were made.Results: Out of the 22 patients with PIH, 20 patients had abnormal umbilical artery S/D ratio and all 22 had abnormal MCA PI. Out of 12 patients with diabetes, 10 had abnormal umbilical artery S/D ratio. All the patients with IUGR had abnormal umbilical artery S/D ratio and abnormal MCA PI.Conclusions: Colour doppler flow velocimetry done repeatedly can predict adverse foetal events with a great degree of accuracy