Heterologous expression of 4α-glucanotransferase: overproduction and properties for industrial applications

2022 ◽  
Vol 38 (2) ◽  
Author(s):  
Santhana Nakapong ◽  
Suthipapun Tumhom ◽  
Jarunee Kaulpiboon ◽  
Piamsook Pongsawasdi
Keyword(s):  
Heterologous Expression ◽  
Industrial Applications

scholarly journals Extracellular production of the engineered thermostable protease pernisine from Aeropyrum pernix K1 in Streptomyces rimosus

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Marko Šnajder ◽  
Andrés Felipe Carrillo Rincón ◽  
Vasilka Magdevska ◽  
Miha Bahun ◽  
Luka Kranjc ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Prion Proteins ◽  
Signal Sequence ◽  
Industrial Applications ◽  
Streptomyces Rimosus ◽  
Extracellular Production ◽  
Industrial Fermentation ◽  
Aeropyrum Pernix ◽  
Processing Step ◽  
Thermophilic Organisms

Abstract Background The thermostable serine protease pernisine originates from the hyperthermophilic Archaeaon Aeropyrum pernix and has valuable industrial applications. Due to its properties, A. pernix cannot be cultivated in standard industrial fermentation facilities. Furthermore, pernisine is a demanding target for heterologous expression in mesophilic heterologous hosts due to the relatively complex processing step involved in its activation. Results We achieved production of active extracellular pernisine in a Streptomyces rimosus host through heterologous expression of the codon-optimised gene by applying step-by-step protein engineering approaches. To ensure secretion of fully active enzyme, the srT signal sequence from the S. rimosus protease was fused to pernisine. To promote correct processing and folding of pernisine, the srT functional cleavage site motif was fused directly to the core pernisine sequence, this way omitting the proregion. Comparative biochemical analysis of the wild-type and recombinant pernisine confirmed that the enzyme produced by S. rimosus retained all of the desired properties of native pernisine. Importantly, the recombinant pernisine also degraded cellular and infectious bovine prion proteins, which is one of the particular applications of this protease. Conclusion Functional pernisine that retains all of the advantageous properties of the native enzyme from the thermophilic host was successfully produced in a S. rimosus heterologous host. Importantly, we achieved extracellular production of active pernisine, which significantly simplifies further downstream procedures and also omits the need for any pre-processing step for its activation. We demonstrate that S. rimosus can be used as an attractive host for industrial production of recombinant proteins that originate from thermophilic organisms.

scholarly journals Green Production and Biotechnological Applications of Cell Wall Lytic Enzymes

2019 ◽  
Vol 9 (23) ◽  
pp. 5012 ◽  
Author(s):  
Benedetti ◽  
Locci ◽  
Gramegna ◽  
Sestili ◽  
Savatin
Keyword(s):  
Cell Wall ◽  
Heterologous Expression ◽  
Energy Demand ◽  
Fossil Fuels ◽  
Dominant Role ◽  
Value Added ◽  
Industrial Applications ◽  
Lytic Enzymes ◽  
Negative Effects ◽  
Cell Wall Lytic Enzymes

: Energy demand is constantly growing, and, nowadays, fossil fuels still play a dominant role in global energy production, despite their negative effects on air pollution and the emission of greenhouse gases, which are the main contributors to global warming. An alternative clean source of energy is represented by the lignocellulose fraction of plant cell walls, the most abundant carbon source on Earth. To obtain biofuels, lignocellulose must be efficiently converted into fermentable sugars. In this regard, the exploitation of cell wall lytic enzymes (CWLEs) produced by lignocellulolytic fungi and bacteria may be considered as an eco-friendly alternative. These organisms evolved to produce a variety of highly specific CWLEs, even if in low amounts. For an industrial use, both the identification of novel CWLEs and the optimization of sustainable CWLE-expressing biofactories are crucial. In this review, we focus on recently reported advances in the heterologous expression of CWLEs from microbial and plant expression systems as well as some of their industrial applications, including the production of biofuels from agricultural feedstock and of value-added compounds from waste materials. Moreover, since heterologous expression of CWLEs may be toxic to plant hosts, genetic strategies aimed in converting such a deleterious effect into a beneficial trait are discussed.

scholarly journals High Level of Spinosad Production in the Heterologous Host Saccharopolyspora erythraea

2016 ◽  
Vol 82 (18) ◽  
pp. 5603-5611 ◽  
Author(s):  
Jun Huang ◽  
Zhen Yu ◽  
Mei-Hong Li ◽  
Ji-Dong Wang ◽  
Hua Bai ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster ◽  
Gene Clusters ◽  
Industrial Applications ◽  
Global Market ◽  
Saccharopolyspora Erythraea ◽  
Market Demand ◽  
Saccharopolyspora Spinosa ◽  
Biosynthesis Gene Cluster ◽  
Content Type

ABSTRACTSpinosad, a highly effective insecticide, has an excellent environmental and mammalian toxicological profile. Global market demand for spinosad is huge and growing. However, after much effort, there has been almost no improvement in the spinosad yield from the original producer,Saccharopolyspora spinosa. Here, we report the heterologous expression of spinosad usingSaccharopolyspora erythraeaas a host. The native erythromycin polyketide synthase (PKS) genes inS. erythraeawere replaced by the assembled spinosad gene cluster through iterative recombination. The production of spinosad could be detected in the recombinant strains containing the whole biosynthesis gene cluster. Both metabolic engineering and UV mutagenesis were applied to further improve the yield of spinosad. The final strain, AT-ES04PS-3007, which could produce spinosad with a titer of 830 mg/liter, has significant potential in industrial applications.IMPORTANCEThis work provides an innovative and promising way to improve the industrial production of spinosad. At the same time, it also describes a successful method of heterologous expression for target metabolites of interest by replacing large gene clusters.

Molecular strategies to increase keratinase production in heterologous expression systems for industrial applications

2021 ◽  
Author(s):  
Radin Shafierul Radin Yahaya ◽  
Yahaya M. Normi ◽  
Lai Yee Phang ◽  
Siti Aqlima Ahmad ◽  
Janna Ong Abdullah ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Industrial Applications ◽  
Expression Systems ◽  
Heterologous Expression Systems

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.

The mensuration of interfaces

1992 ◽  
Vol 50 (1) ◽  
pp. 216-217
Author(s):  
W.M. Stobbs
Keyword(s):  
Electron Microscopy ◽  
50Th Anniversary ◽  
Numerical Data ◽  
Industrial Applications ◽  
Dynamical Theory ◽  
Large Strains ◽  
Reasonable Approach ◽  
Lattice Resolution ◽  
Analogue To Digital ◽  
The Way

I do not have access to the abstracts of the first meeting of EMSA but at this, the 50th Anniversary meeting of the Electron Microscopy Society of America, I have an excuse to consider the historical origins of the approaches we take to the use of electron microscopy for the characterisation of materials. I have myself been actively involved in the use of TEM for the characterisation of heterogeneities for little more than half of that period. My own view is that it was between the 3rd International Meeting at London, and the 1956 Stockholm meeting, the first of the European series , that the foundations of the approaches we now take to the characterisation of a material using the TEM were laid down. (This was 10 years before I took dynamical theory to be etched in stone.) It was at the 1956 meeting that Menter showed lattice resolution images of sodium faujasite and Hirsch, Home and Whelan showed images of dislocations in the XlVth session on “metallography and other industrial applications”. I have always incidentally been delighted by the way the latter authors misinterpreted astonishingly clear thickness fringes in a beaten (”) foil of Al as being contrast due to “large strains”, an error which they corrected with admirable rapidity as the theory developed. At the London meeting the research described covered a broad range of approaches, including many that are only now being rediscovered as worth further effort: however such is the power of “the image” to persuade that the above two papers set trends which influence, perhaps too strongly, the approaches we take now. Menter was clear that the way the planes in his image tended to be curved was associated with the imaging conditions rather than with lattice strains, and yet it now seems to be common practice to assume that the dots in an “atomic resolution image” can faithfully represent the variations in atomic spacing at a localised defect. Even when the more reasonable approach is taken of matching the image details with a computed simulation for an assumed model, the non-uniqueness of the interpreted fit seems to be rather rarely appreciated. Hirsch et al., on the other hand, made a point of using their images to get numerical data on characteristics of the specimen they examined, such as its dislocation density, which would not be expected to be influenced by uncertainties in the contrast. Nonetheless the trends were set with microscope manufacturers producing higher and higher resolution microscopes, while the blind faith of the users in the image produced as being a near directly interpretable representation of reality seems to have increased rather than been generally questioned. But if we want to test structural models we need numbers and it is the analogue to digital conversion of the information in the image which is required.

The influence of confocal microscope design on imaging performance

1993 ◽  
Vol 51 ◽  
pp. 144-145
Author(s):  
C J R Sheppard
Keyword(s):  
Image Quality ◽  
Fluorescence Imaging ◽  
Confocal Microscope ◽  
Industrial Applications ◽  
Three Dimensions ◽  
Design Parameters ◽  
Weak Signals ◽  
Size And Shape ◽  
Imaging Performance ◽  
Fundamental Limitation

The confocal microscope is now widely used in both biomedical and industrial applications for imaging, in three dimensions, objects with appreciable depth. There are now a range of different microscopes on the market, which have adopted a variety of different designs. The aim of this paper is to explore the effects on imaging performance of design parameters including the method of scanning, the type of detector, and the size and shape of the confocal aperture.It is becoming apparent that there is no such thing as an ideal confocal microscope: all systems have limitations and the best compromise depends on what the microscope is used for and how it is used. The most important compromise at present is between image quality and speed of scanning, which is particularly apparent when imaging with very weak signals. If great speed is not of importance, then the fundamental limitation for fluorescence imaging is the detection of sufficient numbers of photons before the fluorochrome bleaches.

Microstructural characterization of sol-gel coating on PET films

1994 ◽  
Vol 52 ◽  
pp. 886-887
Author(s):  
R. T. Chen ◽  
R.A. Norwood
Keyword(s):  
Ion Beam ◽  
Sol Gel ◽  
Industrial Applications ◽  
Hard Coatings ◽  
Nanometer Scale ◽  
Ion Beam Sputtering ◽  
Process Technology ◽  
Refractive Index Measurement ◽  
Organically Modified ◽  
Pet Films

Sol-gel processing has been used to control the structure of a material on a nanometer scale in preparing advanced ceramics and glasses. Film coating using the sol-gel process was also found to be a viable process technology in applications such as optical, porous, antireflection and hard coatings. In this study, organically modified silicate (Ormosil) coatings are applied to PET films for various industrial applications. Sol-gel materials are known to exhibit nanometer scale structures which havepreviously been characterized by small-angle X-ray scattering (SAXS), neutron scattering and light scattering. Imaging of the ultrafine sol-gel structures has also been performed using an ultrahigh resolution replica/TEM technique. The objective of this study was to evaluate the ultrafine structures inthe sol gel coatings using a direct imaging technique: atomic force microscopy (AFM). In addition, correlation of microstructures with processing parameters, coating density and other physical properties will be discussed.The materials evaluated are organically modified silicate coatings on PET film substrates. Refractive index measurement by the prism coupling method was used to assess density of the sol-gel coating.AFM imaging was performed on a Nanoscope III AFM (by Digital Instruments) using constant force mode. Solgel coating samples coated with a thin layer of Ft (by ion beam sputtering) were also examined by STM in order to confirm the structures observed in the contact type AFM. In addition, to compare the previous results, sol-gel powder samples were also prepared by ultrasonication followed by Pt/Au shadowing and examined using a JEOL 100CX TEM.

Electron energy loss near edge structures of intermetallic alloys and grain boundaries in NiAl

1996 ◽  
Vol 54 ◽  
pp. 522-523
Author(s):  
G.A. Botton ◽  
C.J. Humphreys
Keyword(s):  
High Temperature ◽  
Transition Metal ◽  
Energy Loss ◽  
Grain Boundaries ◽  
Industrial Applications ◽  
Edge Structure ◽  
Structural Applications ◽  
Yield Behaviour ◽  
Late Transition ◽  
Metal Aluminides

Transition metal aluminides are of great potential interest for high temperature structural applications. Although these materials exhibit good mechanical properties at high temperature, their use in industrial applications is often limited by their intrinsic room temperature brittleness. Whilst this particular yield behaviour is directly related to the defect structure, the properties of the defects (in particular the mobility of dislocations and the slip system on which these dislocations move) are ultimately determined by the electronic structure and bonding in these materials. The lack of ductility has been attributed, at least in part, to the mixed bonding character (metallic and covalent) as inferred from ab-initio calculations. In this work, we analyse energy loss spectra and discuss the features of the near edge structure in terms of the relevant electronic states in order to compare the predictions on bonding directly with spectroscopic experiments. In this process, we compare spectra of late transition metal (TM) to early TM aluminides (FeAl and TiAl) to assess whether differences in bonding can also be detected. This information is then discussed in terms of bonding changes at grain boundaries in NiAl.

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