Functional and structural features of the oxyanion hole in a thermophilic esterase from Alicyclobacillus acidocaldarius

2007 ◽  
Vol 71 (4) ◽  
pp. 1721-1731 ◽  
Author(s):  
Luigi Mandrich ◽  
Valeria Menchise ◽  
Vincenzo Alterio ◽  
Giuseppina De Simone ◽  
Carlo Pedone ◽  
...  
Keyword(s):  
Structural Features ◽  
Oxyanion Hole ◽  
Thermophilic Esterase ◽  
Alicyclobacillus Acidocaldarius

scholarly journals Alicyclobacillus acidocaldarius Thermophilic Esterase EST2's Activity in Milk and Cheese Models

2006 ◽  
Vol 72 (5) ◽  
pp. 3191-3197 ◽  
Author(s):  
Luigi Mandrich ◽  
Giuseppe Manco ◽  
Mos� Rossi ◽  
Esther Floris ◽  
Tanja Jansen-van den Bosch ◽  
...  
Keyword(s):  
Short Chain Fatty Acid ◽  
Hydrolytic Activity ◽  
Spectrophotometric Assay ◽  
Fatty Acid Esters ◽  
Optimal Conditions ◽  
Ester Synthesis ◽  
Thermophilic Enzyme ◽  
Thermophilic Esterase ◽  
Alicyclobacillus Acidocaldarius ◽  
Acid Esters

ABSTRACT The aim of this work was to investigate the behavior of thermophilic esterase EST2 from Alicyclobacillus acidocaldarius in milk and cheese models. The pure enzyme was used to compare the EST2 hydrolytic activity to the activity of endogenous esterase EstA from Lactococcus lactis. The results indicate that EST2 exhibits 30-fold-higher esterase activity than EstA. As EstA has thioesterase activity, EST2 was assayed for this activity under the optimal conditions determined for EstA (namely, 30�C and pH 7.5). Although it is a thermophilic enzyme, EST2 exhibited eightfold-higher thioesterase activity than EstA with S-methyl thiobutanoate. The abilities of EST2 and EstA to synthesize short-chain fatty acid esters were compared. Two methods were developed to do this. In the first method a spectrophotometric assay was used to monitor the synthesis of esters by the pure enzymes using p-nitrophenol as the alcohol substrate. The synthetic activities were also evaluated under conditions that mimicked those present in milk and/or cheese. The second method involved evaluation of the synthetic abilities of the enzymes when they were directly added to a model cheese matrix. Substantial ester synthesis by EST2 was observed under both conditions. Finally, esterase and thioesterase activities were evaluated in milk using the purified EST2 enzyme and in the model cheese matrix using a strain of L. lactis NZ9000 harboring the EST2 gene and thus overproducing EST2. Both the esterase and thioesterase activities measured in milk and in the cheese matrix were much greater than the activities of the controls.

Enlarging the substrate portfolio of the thermophilic esterase EST2 from Alicyclobacillus acidocaldarius

Extremophiles ◽  
2015 ◽  
Vol 19 (5) ◽  
pp. 1001-1011 ◽  
Author(s):  
Angela Pennacchio ◽  
Luigi Mandrich ◽  
Giuseppe Manco ◽  
Antonio Trincone
Keyword(s):  
Thermophilic Esterase ◽  
Alicyclobacillus Acidocaldarius

Irreversible inhibition of the thermophilic esterase EST2 from Alicyclobacillus acidocaldarius

Extremophiles ◽  
2008 ◽  
Vol 12 (5) ◽  
pp. 719-728 ◽  
Author(s):  
Ferdinando Febbraio ◽  
Sandro Esposito D’Andrea ◽  
Luigi Mandrich ◽  
Luigia Merone ◽  
Mosè Rossi ◽  
...  
Keyword(s):  
Irreversible Inhibition ◽  
Thermophilic Esterase ◽  
Alicyclobacillus Acidocaldarius

Electron microscopy and diffraction studies of polyimides

1983 ◽  
Vol 41 ◽  
pp. 28-29
Author(s):  
O.C. de Hodgins ◽  
K. R. Lawless ◽  
R. Anderson
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Structural Features ◽  
Inert Atmosphere ◽  
Polyimide Films ◽  
Resolution Electron ◽  
The Matrix ◽  
High Resolution Electron Microscope ◽  
Diffraction Patterns ◽  
Polymeric Samples

Commercial polyimide films have shown to be homogeneous on a scale of 5 to 200 nm. The observation of Skybond (SKB) 705 and PI5878 was carried out by using a Philips 400, 120 KeV STEM. The objective was to elucidate the structural features of the polymeric samples. The specimens were spun and cured at stepped temperatures in an inert atmosphere and cooled slowly for eight hours. TEM micrographs showed heterogeneities (or nodular structures) generally on a scale of 100 nm for PI5878 and approximately 40 nm for SKB 705, present in large volume fractions of both specimens. See Figures 1 and 2. It is possible that the nodulus observed may be associated with surface effects and the structure of the polymers be regarded as random amorphous arrays. Diffraction patterns of the matrix and the nodular areas showed different amorphous ring patterns in both materials. The specimens were viewed in both bright and dark fields using a high resolution electron microscope which provided magnifications of 100,000X or more on the photographic plates if desired.

Skeletal elements of crinoid echinoderms: High-resolution structural and microanalytical results

1983 ◽  
Vol 41 ◽  
pp. 194-195
Author(s):  
D. F. Blake ◽  
L. F. Allard ◽  
D. R. Peacor
Keyword(s):  
High Resolution ◽  
Marine Invertebrates ◽  
Sea Urchins ◽  
Structural Features ◽  
Sea Stars ◽  
High Resolution Tem ◽  
Relationship Of ◽  
The Relationship ◽  
Insight Into

Echinodermata is a phylum of marine invertebrates which has been extant since Cambrian time (c.a. 500 m.y. before the present). Modern examples of echinoderms include sea urchins, sea stars, and sea lilies (crinoids). The endoskeletons of echinoderms are composed of plates or ossicles (Fig. 1) which are with few exceptions, porous, single crystals of high-magnesian calcite. Despite their single crystal nature, fracture surfaces do not exhibit the near-perfect {10.4} cleavage characteristic of inorganic calcite. This paradoxical mix of biogenic and inorganic features has prompted much recent work on echinoderm skeletal crystallography. Furthermore, fossil echinoderm hard parts comprise a volumetrically significant portion of some marine limestones sequences. The ultrastructural and microchemical characterization of modern skeletal material should lend insight into: 1). The nature of the biogenic processes involved, for example, the relationship of Mg heterogeneity to morphological and structural features in modern echinoderm material, and 2). The nature of the diagenetic changes undergone by their ancient, fossilized counterparts. In this study, high resolution TEM (HRTEM), high voltage TEM (HVTEM), and STEM microanalysis are used to characterize tha ultrastructural and microchemical composition of skeletal elements of the modern crinoid Neocrinus blakei.

Study of the Molecular Architecture of Keratin Filaments by Electron Microscopy

1982 ◽  
Vol 40 ◽  
pp. 118-119
Author(s):  
U. Aebi ◽  
P. Rew ◽  
T.-T. Sun
Keyword(s):  
Electron Microscopy ◽  
Structural Organization ◽  
Cell Types ◽  
Structural Features ◽  
Molecular Architecture ◽  
The Past ◽  
Keratin Filaments ◽  
Different Types ◽  
10 Nm Filaments ◽  
Different Cell Types

Various types of intermediate-sized (10-nm) filaments have been found and described in many different cell types during the past few years. Despite the differences in the chemical composition among the different types of filaments, they all yield common structural features: they are usually up to several microns long and have a diameter of 7 to 10 nm; there is evidence that they are made of several 2 to 3.5 nm wide protofilaments which are helically wound around each other; the secondary structure of the polypeptides constituting the filaments is rich in ∞-helix. However a detailed description of their structural organization is lacking to date.

Measurement and Reduction of Radiation Damage in Frozen Hydrated Crystalline Specimens

1978 ◽  
Vol 36 (3) ◽  
pp. 70-77 ◽  
Author(s):  
R.M. Glaeser ◽  
S.B. Hayward
Keyword(s):  
Diffraction Pattern ◽  
Structural Information ◽  
Structural Disorder ◽  
Structural Features ◽  
Biological Macromolecules ◽  
Diffraction Intensity ◽  
Object Structure ◽  
Specimen Material ◽  
Unit Cells ◽  
Identical Unit

Highly ordered or crystalline biological macromolecules become severely damaged and structurally disordered after a brief electron exposure. Evidence that damage and structural disorder are occurring is clearly given by the fading and eventual disappearance of the specimen's electron diffraction pattern. The fading and disappearance of sharp diffraction spots implies a corresponding disappearance of periodic structural features in the specimen. By the same token, there is a oneto- one correspondence between the disappearance of the crystalline diffraction pattern and the disappearance of reproducible structural information that can be observed in the images of identical unit cells of the object structure. The electron exposures that result in a significant decrease in the diffraction intensity will depend somewhat upon the resolution (Bragg spacing) involved, and can vary considerably with the chemical makeup and composition of the specimen material.

Some Structural Features of Metaphase Chromosomes of Mice and Men

1967 ◽  
Vol 25 ◽  
pp. 190-191
Author(s):  
Godfrey C. Hoskins ◽  
Betty B. Hoskins
Keyword(s):  
Electron Microscopy ◽  
Electron Micrographs ◽  
Structural Features ◽  
Metaphase Chromosomes ◽  
The Future ◽  
Of Mice And Men ◽  
Mouse Cells ◽  
Human And Mouse

Metaphase chromosomes from human and mouse cells in vitro are isolated by micrurgy, fixed, and placed on grids for electron microscopy. Interpretations of electron micrographs by current methods indicate the following structural features.Chromosomal spindle fibrils about 200Å thick form fascicles about 600Å thick, wrapped by dense spiraling fibrils (DSF) less than 100Å thick as they near the kinomere. Such a fascicle joins the future daughter kinomere of each metaphase chromatid with those of adjacent non-homologous chromatids to either side. Thus, four fascicles (SF, 1-4) attach to each metaphase kinomere (K). It is thought that fascicles extend from the kinomere poleward, fray out to let chromosomal fibrils act as traction fibrils against polar fibrils, then regroup to join the adjacent kinomere.

Observations on the Structure of Ta2O5

1972 ◽  
Vol 30 ◽  
pp. 540-541
Author(s):  
P. S. Kotval ◽  
C. J. Dewit
Keyword(s):  
Electron Microscope ◽  
Structural Features ◽  
Amorphous Structure ◽  
Polished Surface ◽  
Distilled Water ◽  
Anodic Films ◽  
Amorphous Films ◽  
Beam Heating ◽  
Diffraction Patterns ◽  
Hexagonal Arrays

The structure of Ta2O5 has been described in the literature in several different crystallographic forms with varying unit cell lattice parameters. Earlier studies on films of Ta2O5 produced by anodization of tantalum have revealed structural features which are not consistent with the parameters of “bulk” Ta2O5 crystalsFilms of Ta2O5 were prepared by anodizing a well-polished surface of pure tantalum sheet. The anodic films were floated off in distilled water, collected on grids, dried and directly examined in the electron microscope. In all cases the films were found to exhibit diffraction patterns representative of an amorphous structure. Using beam heating in the electron microscope, recrystallization of the amorphous films can be accomplished as shown in Fig. 1. As suggested by earlier work, the recrystallized regions exhibit diffraction patterns which consist of hexagonal arrays of main spots together with subsidiary rows of super lattice spots which develop as recrystallization progresses (Figs. 2a and b).

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