Isolation and partial characterization of the parasporal body of Bacillus popillae

1978 ◽  
Vol 24 (12) ◽  
pp. 1557-1561 ◽  
Author(s):  
B. A. Weiner

The sporangial wall of Bacillus popilliae encloses a spore and parasporal body and is very resistant to breakage. High-energy sonication can disrupt this envelope and release free spores and parasporal bodies which retain refractility. Parasporal bodies, free spores, and sporangial debris were separated by centrifugation in linear sucrose gradients. Purified parasporal bodies are proteinaceous. Of the 17 amino acids detected, glutamic and aspartic acids are highest in concentration and account for the low isoelectric point of 4.5; a lipid moiety was not found. Parasporal bodies are soluble in 0.1 N NaOH, 8 M guanidine hydrochloride, 8 M urea, or alkaline thioglycollate. Paper electrophoresis at 3000 V separated soluble parasporal protein into three cathodic components. Solubilized or whole intact parasporal bodies fed to third-instar larvae of the Japanese beetle were not detectably toxic. However, solubilized parasporal protein was twice as toxic as the intact parasporal bodies when injected into the hemocoel.

1989 ◽  
Vol 44 (9-10) ◽  
pp. 749-753 ◽  
Author(s):  
Daniela Woitzik ◽  
Roland Dierstein ◽  
Jürgen Weckesser

The lipoprotein-peptidoglycan complex (rigid layer) from Rhodobacter sphaeroides was isolated. Treatment of the complex with N.O-diacetyl-muramidase cleaved off from the rigid layer the lipoprotein moiety with covalently bound peptidoglycan-fragments. The lipoprotein with the bound fragments showed a broad single band (Mr about 19,000) on SDS-polyacrylamide gels and had an isoelectric point of about 5.6. There was no serological cross-reaction with the rigid layer of Escherichia coli K12. The lipid moiety of lipoprotein with residual amino acids (mainly Gly, Ser, Gly, Asp) was obtained by Pronase E treatment of the rigid layer and chloroform -methanol extraction. It was free from phosphate and contained amide- and esterbound 18:1 and esterbound 16:0 and 18:0. The isolated peptidoglycan, after enzymatical cleavage from the lipoprotein, had a chemical composition indicating Aly-type structure. Comparable studies, performed with Rhodobacter capsulatus 37b4 resulted in essentially similar results for rigid layer, lipoprotein and peptidoglycan compositions.


1972 ◽  
Vol 127 (3) ◽  
pp. 489-502 ◽  
Author(s):  
I. M. Bartley ◽  
B. Hodgson ◽  
J. S. Walker ◽  
G. Holme

Cell extracts from Bacillus brevis (A.T.C.C. 10068), grown with various media, incorporated certain 14C-labelled amino acids that are normally components of tyrothricin into material that was extracted by ethanol from the precipitate formed by adding acid. When this material was separated by paper and silica-gel thin-layer chromatography and paper electrophoresis 14C was located in those regions that also contained gramicidin and tyrocidine. From a study of the properties of the system responsible for the incorporation it was deduced that non-tyrothricin materials were present. It was shown that the methods normally used to characterize tyrothricin do not adequately distinguish between tyrothricin and non-tyrothricin materials. However, a method for separating these materials was devised. This involved elution with ethanol from columns of acid alumina followed by gel filtration on Sephadex LH-20 with dimethylformamide–water solvent. The behaviour of gramicidin and tyrocidine on the Sephadex LH-20 column was examined, and it was concluded that the separation was not caused simply by gel filtration of unassociated molecules. Also, tyrocidine molecules with different amino acid compositions seemed to have different affinities for the Sephadex LH-20 column.


2016 ◽  
Author(s):  
Antonio Lanzirotti ◽  
◽  
Stephen R. Sutton ◽  
Matt Newville ◽  
Jeffrey P. Fitts ◽  
...  

1984 ◽  
Vol 49 (8) ◽  
pp. 1846-1853 ◽  
Author(s):  
Karel Hauzer ◽  
Tomislav Barth ◽  
Linda Servítová ◽  
Karel Jošt

A post-proline endopeptidase (EC 3.4.21.26) was isolated from pig kidneys using a modified method described earlier. The enzyme was further purified by ion exchange chromatography on DEAE-Sephacel. The final product contained about 95% of post-proline endopeptidase. The enzyme molecule consisted of one peptide chain with a relative molecular mass of 65 600 to 70 000, containing a large proportion of acidic and alifatic amino acids (glutamic acid, aspartic acid and leucine) and the N-terminus was formed by aspartic acid or asparagine. In order to prevent losses of enzyme activity, thiol compounds has to be added.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sonja Christiane Bäßler ◽  
Ákos Kenéz ◽  
Theresa Scheu ◽  
Christian Koch ◽  
Ulrich Meyer ◽  
...  

AbstractMetabolic consequences of an energy and protein rich diet can compromise metabolic health of cattle by promoting a pro-inflammatory phenotype. Laminitis is a common clinical sign, but affected metabolic pathways, underlying pathophysiology and causative relationships of a systemic pro-inflammatory phenotype are unclear. Therefore, the aim of this study was to elucidate changes in metabolome profiles of 20 months old Holstein bulls fed a high energy and protein diet and to identify novel metabolites and affected pathways, associated with diet-related laminitis. In a randomized controlled feeding trial using bulls fed a high energy and protein diet (HEP; metabolizable energy [ME] intake 169.0 ± 1.4 MJ/day; crude protein [CP] intake 2.3 ± 0.02 kg/day; calculated means ± SEM; n = 15) versus a low energy and protein diet (LEP; ME intake 92.9 ± 1.3 MJ/day; CP intake 1.0 ± 0.01 kg/day; n = 15), wide ranging effects of HEP diet on metabolism were demonstrated with a targeted metabolomics approach using the AbsoluteIDQ p180 kit (Biocrates Life Sciences). Multivariate statistics revealed that lower concentrations of phosphatidylcholines and sphingomyelins and higher concentrations of lyso-phosphatidylcholines, branched chain amino acids and aromatic amino acids were associated with an inflammatory state of diet-related laminitis in Holstein bulls fed a HEP diet. The latter two metabolites share similarities with changes in metabolism of obese humans, indicating a conserved pathophysiological role. The observed alterations in the metabolome provide further explanation on the underlying metabolic consequences of excessive dietary nutrient intake.


Polymers ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 1971
Author(s):  
Lihua Ye ◽  
Muhammad Muzamal Ashfaq ◽  
Aiping Shi ◽  
Syyed Adnan Raheel Shah ◽  
Yefan Shi

In this research, the aim relates to the material characterization of high-energy lithium-ion pouch cells. The development of appropriate model cell behavior is intended to simulate two scenarios: the first is mechanical deformation during a crash and the second is an internal short circuit in lithium-ion cells during the actual effect scenarios. The punch test has been used as a benchmark to analyze the effects of different state of charge conditions on high-energy lithium-ion battery cells. This article explores the impact of three separate factors on the outcomes of mechanical punch indentation experiments. The first parameter analyzed was the degree of prediction brought about by experiments on high-energy cells with two different states of charge (greater and lesser), with four different sizes of indentation punch, from the cell’s reaction during the indentation effects on electrolyte. Second, the results of the loading position, middle versus side, are measured at quasi-static speeds. The third parameter was the effect on an electrolyte with a different state of charge. The repeatability of the experiments on punch loading was the last test function analyzed. The test results of a greater than 10% state of charge and less than 10% state of charge were compared to further refine and validate this modeling method. The different loading scenarios analyzed in this study also showed great predictability in the load-displacement reaction and the onset short circuit. A theoretical model of the cell was modified for use in comprehensive mechanical deformation. The overall conclusion found that the loading initiating the cell’s electrical short circuit is not instantaneously instigated and it is subsequently used to process the development of a precise and practical computational model that will reduce the chances of the internal short course during the crash.


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