scholarly journals A three-component monooxygenase from Rhodococcus wratislaviensis may expand industrial applications of bacterial enzymes

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Makoto Hibi ◽  
Dai Fukuda ◽  
Chihiro Kenchu ◽  
Masutoshi Nojiri ◽  
Ryotaro Hara ◽  
...  
Keyword(s):  
Proteome Analysis ◽  
Initial Step ◽  
Industrial Applications ◽  
Crystal Structure Analysis ◽  
Methyl Groups ◽  
Monooxygenase System ◽  
Oxidative Reactivity ◽  
Diiron Enzymes ◽  
Amidohydrolase Superfamily ◽  
Rhodococcus Wratislaviensis

AbstractThe high-valent iron-oxo species formed in the non-heme diiron enzymes have high oxidative reactivity and catalyze difficult chemical reactions. Although the hydroxylation of inert methyl groups is an industrially promising reaction, utilizing non-heme diiron enzymes as such a biocatalyst has been difficult. Here we show a three-component monooxygenase system for the selective terminal hydroxylation of α-aminoisobutyric acid (Aib) into α-methyl-D-serine. It consists of the hydroxylase component, AibH1H2, and the electron transfer component. Aib hydroxylation is the initial step of Aib catabolism in Rhodococcus wratislaviensis C31-06, which has been fully elucidated through a proteome analysis. The crystal structure analysis revealed that AibH1H2 forms a heterotetramer of two amidohydrolase superfamily proteins, of which AibHm2 is a non-heme diiron protein and functions as a catalytic subunit. The Aib monooxygenase was demonstrated to be a promising biocatalyst that is suitable for bioprocesses in which the inert C–H bond in methyl groups need to be activated.

scholarly journals Transcriptome and proteome analysis suggest enhanced photosynthesis in tetraploid Liriodendron sino-americanum

2021 ◽  
Author(s):  
Tingting Chen ◽  
Yu Sheng ◽  
Zhaodong Hao ◽  
Xiaofei Long ◽  
Fangfang Fu ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Protein Complexes ◽  
Proteome Analysis ◽  
Tree Height ◽  
Photosynthetic Electron Transport ◽  
Industrial Applications ◽  
Chlorophyll Protein Complexes ◽  
Chlorophyll Protein ◽  
Photosynthesis Genes ◽  
Underlying Mechanisms

Abstract Polyploidy generally provides an advantage in phenotypic variation and growth vigor. However, the underlying mechanisms remain poorly understood. The tetraploid L. sino-americanum exhibits altered morphology compared to its diploid counterpart, including larger, thicker and deeper green leaves, bigger stomata, thicker stems and increased tree height. Such characteristics can be useful in ornamental and industrial applications. To elucidate the molecular mechanisms behind this variation, we performed a comparative transcriptome and proteome analysis. Our transcriptome data indicated that some photosynthesis genes and pathways were differentially altered and enriched in tetraploid L. sino-americanum, mainly related to F-type ATPase, the cytochrome b6/f complex, photosynthetic electron transport, the light harvesting chlorophyll protein complexes, photosystem I and II. Most of the differentially expressed proteins we could identify are also involved in photosynthesis. Our physiological results showed that tetraploids have an enhanced photosynthetic capacity, concomitant with great levels of sugar and starch in leaves. This suggests that tetraploid L. sino-americanum might experience comprehensive transcriptome reprogramming of genes related to photosynthesis. This study has especially emphasized molecular changes involved in photosynthesis that accompany polyploidy, and provides a possible explanation for the altered phenotype of polyploidy plants in comparison to their diploid form.

Three-Dimensional High-Resolution Optical/X-Ray Stereoscopic Tracking Velocimetry

2004 ◽  
Author(s):  
David J. Lee ◽  
Soyoung S. Cha ◽  
Narayanan Ramachandran
Keyword(s):  
Three Dimensional ◽  
Initial Step ◽  
Industrial Applications ◽  
Materials Processing ◽  
Current Status ◽  
Motion Velocity ◽  
Processing Efficiency ◽  
X Ray ◽  
Fluid Physics ◽  
Component Velocity

Measurement of three-dimensional (3-D) three-component velocity fields is of great importance in a variety of research and industrial applications for understanding materials processing, fluid physics, and strain/displacement measurements. The 3-D experiments in these fields most likely inhibit the use of conventional techniques, which are based only on planar and optically-transparent-field observation. Here, we briefly review the current status of 3-D diagnostics for motion/velocity detection, for both optical and x-ray systems. As an initial step for providing 3-D capabilities, we have developed stereoscopic tracking velocimetry (STV) to measure 3-D flow/deformation through optical observation. The STV is advantageous in system simplicity, for continually observing 3-D phenomena in near real-time. In an effort to enhance the data processing through automation and to avoid the confusion in tracking numerous markers or particles, artificial neural networks are employed to incorporate human intelligence. Our initial optical investigations have proven the STV to be a very viable candidate for reliably measuring 3-D flow motions. With previous activities are focused on improving the processing efficiency, overall accuracy, and automation based on the optical system, the current efforts is directed to the concurrent expansion to the x-ray system for broader experimental applications.

The stereochemistry of B -diketo complexes with trim ethylplatinum iv II. The crystal structure of ethyl(trim ethylplatini-)- acetoacetate

1960 ◽  
Vol 254 (1277) ◽  
pp. 218-228 ◽  
Keyword(s):  
Crystal Structure ◽  
Three Dimensional ◽  
Carbonyl Oxygen ◽  
Crystal Structure Analysis ◽  
Methyl Groups ◽  
Methylene Carbon ◽  
X Ray ◽  
Methylene Carbon Atom ◽  
Critical Account ◽  
Active Methylene

A three-dimensional X-ray crystal structure analysis of the complex between trimethyl platinum and ethylacetoacetate, (CH 3 ) 3 Pt CH 3 . CO. CH. CO OC 2 H 5 , has shown that the molecule is dimeric. In a monoclinic unit cell (a = 8.83, b = 14.12, c = 9.30 Å, β = 95°, space group P2 1 / c ) there are two centrosymmetrical dimeric molecules in which each platinum atom is octahedrally co-ordinated by three methyl groups, in the cis configuration, by the two carbonyl oxygen atoms of one β -ketoester and by the central, or ‘active methylene’ carbon atom of the other β -ketoester in the dimer. The structure is thus essentially the same as that of trimethyl 4:6-dioxononyl platinum (part I) and the result shows that complex formation via a tridentate β -diketo system is preferred to co-ordination through an ester oxygen. A critical account is given of the criteria used to judge the correctness of the results.

scholarly journals Automatic support control of an upper body exoskeleton — Method and validation using the Stuttgart Exo-Jacket

2020 ◽  
Vol 1 ◽  
Author(s):  
Raphael Singer ◽  
Christophe Maufroy ◽  
Urs Schneider
Keyword(s):  
Upper Limb ◽  
Cost Efficiency ◽  
Muscle Activation ◽  
High Reliability ◽  
Initial Step ◽  
High Accuracy ◽  
Industrial Applications ◽  
Upper Body ◽  
Upper Limbs ◽  
Control Approach

Abstract Although passive occupational exoskeletons alleviate worker physical stresses in demanding postures (e.g., overhead work), they are unsuitable in many other applications because of their lack of flexibility. Active exoskeletons that are able to dynamically adjust the delivered support are required. However, the automatic control of support provided by the exoskeleton is still a largely unsolved challenge in many applications, especially for upper limb occupational exoskeletons, where no practical and reliable approach exists. For this type of exoskeletons, a novel support control approach for lifting and carrying activities is presented here. As an initial step towards a full-fledged automatic support control (ASC), the present article focusses on the functionality of estimating the onset of user’s demand for support. In this way, intuitive behavior should be made possible. The combination of movement and muscle activation signals of the upper limbs is expected to enable high reliability, cost efficiency, and compatibility for use in industrial applications. The functionality consists of two parts: a preprocessing—the motion interpretation—and the support detection itself. Both parts were trained with different subjects, who had to move objects. The functionality was validated both in the cases of (A) an unknown subject performing known tasks and (B) a known subject performing unknown tasks. The functionality showed sound results as it achieved a high accuracy ( $$ 95\% $$ ) in training. In addition, the first validation results showed that this functionality is useful for integration in an appropriately adapted ASC and can then enable comfortable working.

Synthesis, Crystal Structure and Magnetism of a Macrocyclic Binuclear Dicopper(II) Amino Alcohol Complex From a Metal Directed Reaction Involving Formaldehyde and Nitroethane

1990 ◽  
Vol 43 (1) ◽  
pp. 69 ◽  
Author(s):  
P Comba ◽  
GA Lawrance ◽  
TM Manning ◽  
A Markiewicz ◽  
KS Murray ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Binuclear Complex ◽  
Room Temperature ◽  
Energy Gap ◽  
Copper Ions ◽  
Crystal Structure Analysis ◽  
Methyl Groups ◽  
Monoclinic Space Group ◽  
Triplet Energy ◽  
Perchlorate Salt

Condensation of the bis (1,5-diaminopentan-3-ol) dicopper (II) ion with formaldehyde and nitroethane in basic methanol yields the macromonocyclic ligand 3,13-dimethyl-3,13-dinitro-1,5,11,15-tetraazaeicosane-8,18-diol as the dicopper (II) complex. The perchlorate salt of the binuclear complex is very strongly antiferromagnetically coupled, with the magnetic properties measured from 10-300 K identifying a singlet-triplet energy gap of -860 cm-1 and a magnetic moment at room temperature of 0.48 BM. The discrete, binuclear nature of the complex was confirmed by a crystal structure analysis of the nitrite salt, which crystallizes in the monoclinic space group C2/c, a 14.789(6), b 14.560(6), c 12.759(9)Ǻ, β 99.65(4)°, and consists of the centrosymmetric macrocycle containing two copper ions each coordinated by two secondary nitrogen donors and two (shared) RO- groups, with water and nitrite oxygens occupying axial sites. The macrocycle donors and the copper ions are essentially coplanar. The Cu…Cu distance is 3.031(1)Ǻ, with the Cu-O--Cu angle 103.6(2)°. The nitro (and methyl) groups at extremities of the macrocycle are in anti dispositions.

scholarly journals Development of New Antiproliferative Compound against Human Tumor Cells from the Marine Microalgae Nannochloropsis gaditana by Applied Proteomics

2020 ◽  
Vol 22 (1) ◽  
pp. 96
Author(s):  
Rafael Carrasco-Reinado ◽  
Almudena Escobar-Niño ◽  
Carlos Fajardo ◽  
Ines M. Morano ◽  
Francisco Amil-Ruiz ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Protein Interactions ◽  
Human Tumor ◽  
Colon Adenocarcinoma ◽  
Proteome Analysis ◽  
Industrial Applications ◽  
Alternative Source ◽  
Nannochloropsis Gaditana ◽  
Bioinformatic Tools

Proteomics is a crucial tool for unravelling the molecular dynamics of essential biological processes, becoming a pivotal technique for basic and applied research. Diverse bioinformatic tools are required to manage and explore the huge amount of information obtained from a single proteomics experiment. Thus, functional annotation and protein–protein interactions are evaluated in depth leading to the biological conclusions that best fit the proteomic response in the system under study. To gain insight into potential applications of the identified proteins, a novel approach named “Applied Proteomics” has been developed by comparing the obtained protein information with the existing patents database. The development of massive sequencing technology and mass spectrometry (MS/MS) improvements has allowed the application of proteomics nonmodel microorganisms, which have been deeply described as a novel source of metabolites. Between them, Nannochloropsis gaditana has been pointed out as an alternative source of biomolecules. Recently, our research group has reported the first complete proteome analysis of this microalga, which was analysed using the applied proteomics concept with the identification of 488 proteins with potential industrial applications. To validate our approach, we selected the UCA01 protein from the prohibitin family. The recombinant version of this protein showed antiproliferative activity against two tumor cell lines, Caco2 (colon adenocarcinoma) and HepG-2 (hepatocellular carcinoma), proving that proteome data have been transformed into relevant biotechnological information. From Nannochloropsis gaditana has been developed a new tool against cancer—the protein named UCA01. This protein has selective effects inhibiting the growth of tumor cells, but does not show any effect on control cells. This approach describes the first practical approach to transform proteome information in a potential industrial application, named “applied proteomics”. It is based on a novel bioalgorithm, which is able to identify proteins with potential industrial applications. From hundreds of proteins described in the proteome of N. gaditana, the bioalgorithm identified over 400 proteins with potential uses; one of them was selected as UCA01, “in vitro” and its potential was demonstrated against cancer. This approach has great potential, but the applications are potentially numerous and undefined.

scholarly journals Glycine Betaine Monooxygenase, an Unusual Rieske-Type Oxygenase System, Catalyzes the Oxidative N -Demethylation of Glycine Betaine in Chromohalobacter salexigens DSM 3043

2018 ◽  
Vol 84 (13) ◽  
Author(s):  
Ya-Hui Shao ◽  
Li-Zhong Guo ◽  
Yu-Qing Zhang ◽  
Hao Yu ◽  
Bai-Suo Zhao ◽  
...  
Keyword(s):  
Glycine Betaine ◽  
Biochemical Characterization ◽  
Initial Step ◽  
Degradation Pathway ◽  
Nuclear Magnetic Resonance Analysis ◽  
Monooxygenase System ◽  
Flavin Reductase ◽  
Prosthetic Group ◽  
Content Type ◽  
Chromohalobacter Salexigens

ABSTRACT Although some bacteria, including Chromohalobacter salexigens DSM 3043, can use glycine betaine (GB) as a sole source of carbon and energy, little information is available about the genes and their encoded proteins involved in the initial step of the GB degradation pathway. In the present study, the results of conserved domain analysis, construction of in-frame deletion mutants, and an in vivo functional complementation assay suggested that the open reading frames Csal_1004 and Csal_1005, designated bmoA and bmoB , respectively, may act as the terminal oxygenase and the ferredoxin reductase genes in a novel Rieske-type oxygenase system to convert GB to dimethylglycine in C. salexigens DSM 3043. To further verify their function, BmoA and BmoB were heterologously overexpressed in Escherichia coli , and 13 C nuclear magnetic resonance analysis revealed that dimethylglycine was accumulated in E. coli BL21(DE3) expressing BmoAB or BmoA. In addition, His-tagged BmoA and BmoB were individually purified to electrophoretic homogeneity and estimated to be a homotrimer and a monomer, respectively. In vitro biochemical analysis indicated that BmoB is an NADH-dependent flavin reductase with one noncovalently bound flavin adenine dinucleotide (FAD) as its prosthetic group. In the presence of BmoB, NADH, and flavin, BmoA could aerobically degrade GB to dimethylglycine with the concomitant production of formaldehyde. BmoA exhibited strict substrate specificity for GB, and its demethylation activity was stimulated by Fe 2+ . Phylogenetic analysis showed that BmoA belongs to group V of the Rieske nonheme iron oxygenase (RO) family, and all the members in this group were able to use quaternary ammonium compounds as substrates. IMPORTANCE GB is widely distributed in nature. In addition to being accumulated intracellularly as a compatible solute to deal with osmotic stress, it can be utilized by many bacteria as a source of carbon and energy. However, very limited knowledge is presently available about the molecular and biochemical mechanisms for the initial step of the aerobic GB degradation pathway in bacteria. Here, we report the molecular and biochemical characterization of a novel two-component Rieske-type monooxygenase system, GB monooxygenase (BMO), which is responsible for oxidative demethylation of GB to dimethylglycine in C. salexigens DSM 3043. The results gained in this study extend our knowledge on the catalytic reaction of microbial GB degradation to dimethylglycine.

scholarly journals Substrate and Inhibitor Spectra of Ethylbenzene Dehydrogenase: Perspectives on Application Potential and Catalytic Mechanism

2012 ◽  
Vol 78 (18) ◽  
pp. 6475-6482 ◽  
Author(s):  
Daniel Knack ◽  
Corina Hagel ◽  
Maciej Szaleniec ◽  
Agnieszka Dudzik ◽  
Aleksander Salwinski ◽  
...  
Keyword(s):  
Catalytic Mechanism ◽  
Initial Step ◽  
Industrial Applications ◽  
Calculation Model ◽  
Kinetic Properties ◽  
Chemical Calculation ◽  
Substrate Analogs ◽  
New Process ◽  
Application Potential ◽  
Apparent Kinetic Parameters

ABSTRACTEthylbenzene dehydrogenase (EbDH) catalyzes the initial step in anaerobic degradation of ethylbenzene in denitrifying bacteria, namely, the oxygen-independent hydroxylation of ethylbenzene to (S)-1-phenylethanol. In our study we investigate the kinetic properties of 46 substrate analogs acting as substrates or inhibitors of the enzyme. The apparent kinetic parameters of these compounds give important insights into the function of the enzyme and are consistent with the predicted catalytic mechanism based on a quantum chemical calculation model. In particular, the existence of the proposed substrate-derived radical and carbocation intermediates is substantiated by the formation of alternative dehydrogenated and hydroxylated products from some substrates, which can be regarded as mechanistic models. In addition, these results also show the surprisingly high diversity of EbDH in hydroxylating different kinds of alkylaromatic and heterocyclic compounds to the respective alcohols. This may lead to attractive industrial applications of ethylbenzene dehydrogenase for a new process of producing alcohols via hydroxylation of the corresponding aromatic hydrocarbons rather than the customary procedure of reducing the corresponding ketones.

Preparation of Electron Transparent Metal-Matrix Composites

1968 ◽  
Vol 26 ◽  
pp. 334-335 ◽  
Author(s):  
M. R. Pinnel ◽  
A. Lawley
Keyword(s):  
Stainless Steel ◽  
Load Transfer ◽  
Volume Fraction ◽  
Steel Wire ◽  
Initial Step ◽  
Interfacial Region ◽  
Matrix Composites ◽  
Specific Test ◽  
The Matrix ◽  
The One

Numerous phenomenological descriptions of the mechanical behavior of composite materials have been developed. There is now an urgent need to study and interpret deformation behavior, load transfer, and strain distribution, in terms of micromechanisms at the atomic level. One approach is to characterize dislocation substructure resulting from specific test conditions by the various techniques of transmission electron microscopy. The present paper describes a technique for the preparation of electron transparent composites of aluminum-stainless steel, such that examination of the matrix-fiber (wire), or interfacial region is possible. Dislocation substructures are currently under examination following tensile, compressive, and creep loading. The technique complements and extends the one other study in this area by Hancock.The composite examined was hot-pressed (argon atmosphere) 99.99% aluminum reinforced with 15% volume fraction stainless steel wire (0.006″ dia.).Foils were prepared so that the stainless steel wires run longitudinally in the plane of the specimen i.e. the electron beam is perpendicular to the axes of the wires. The initial step involves cutting slices ∼0.040″ in thickness on a diamond slitting wheel.

The Application of Ultra-Thin Sectioning Techniques to Non-Biological Samples

1968 ◽  
Vol 26 ◽  
pp. 332-333
Author(s):  
C. F. Oster
Keyword(s):  
Biological Sciences ◽  
Epoxy Resin ◽  
Cross Sections ◽  
Biological Samples ◽  
Industrial Applications ◽  
Photographic Film ◽  
Silver Particles ◽  
Thin Sectioning ◽  
Embedding Medium

Although ultra-thin sectioning techniques are widely used in the biological sciences, their applications are somewhat less popular but very useful in industrial applications. This presentation will review several specific applications where ultra-thin sectioning techniques have proven invaluable.The preparation of samples for sectioning usually involves embedding in an epoxy resin. Araldite 6005 Resin and Hardener are mixed so that the hardness of the embedding medium matches that of the sample to reduce any distortion of the sample during the sectioning process. No dehydration series are needed to prepare our usual samples for embedding, but some types require hardening and staining steps. The embedded samples are sectioned with either a prototype of a Porter-Blum Microtome or an LKB Ultrotome III. Both instruments are equipped with diamond knives.In the study of photographic film, the distribution of the developed silver particles through the layer is important to the image tone and/or scattering power. Also, the morphology of the developed silver is an important factor, and cross sections will show this structure.

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