Low-cost agro-industrial sources as a substrate for the production of L-asparaginase using filamentous fungi

Five strains of filamentous fungi (Aspergillus niger strains NRRL 3122 and T0005007-2, Aspergillus oryzae CCT 3940, Aspergillus awamori NRRL 3112 and a Trichoderma sp.) were compared for their capacity to produce endo-polygalacturonase (endo-PG) in solid state fermentation. Maximum pectinolytic activity was reached in 72 h of growth, the best two fungal strains being A. niger T0005007-2 and A. oryzae CCT 3940. Three types of commercial purified pectin and four of unprocessed pectin (tangerine, orange, Tahiti lime and sweet lime rind) were used to assess the effect of pectin on the production of endo-PG by A. niger T0005007-2. Maximum pectinolytic activity was achieved using 6 and 10% (w/w) of purified pectin as inducer. Depending on the origin of the commercial pectin used as inducer, maximum endo-PG levels varied from 223 to 876 units per gram of dry medium (one endo-PG unit (U) was defined as the quantity of enzyme which caused a reduction in viscosity of 50% in a 1% w/v solution of pectin in 30 min), indicating that care should be taken when choosing this component of the medium. When the crude pectins were used as inducers at the same concentration as purified pectin, maximum endo-PG activities were 250-300 units/g. However, by increasing the amount of Tahiti lime rind to 50% (w/w) maximum endo-PG was 919 U/g, thus opening up the possibility of a low cost medium for endo-PG production.

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Intron Retention Coupled with Nonsense-Mediated Decay is Involved in Cellulase Biosynthesis in Cellulolytic Fungi

10.21203/rs.3.rs-1214111/v1 ◽

2022 ◽

Author(s):

Yichen Gao ◽

Ai-Ping Pang ◽

Leyao Ma ◽

Haiyan Wang ◽

Samran Durrani ◽

...

Keyword(s):

Gene Expression ◽

Filamentous Fungi ◽

Regulatory Networks ◽

Intron Retention ◽

Low Cost ◽

Cellulase Production ◽

Mrna Levels ◽

Nonsense Mediated Decay ◽

Tor Pathway ◽

Regulatory Systems

Abstract Background Knowledge on regulatory networks associated with cellulase biosynthesis is prerequisite for exploitation of such regulatory systems in ehancing cellulase production with low cost. The biological functions of intron retention (IR) and nonsense-mediated mRNA decay (NMD) in filamentous fungi is lack of study, let alone their roles in cellulase biosynthesis. Result We found that major cellulase genes (cel7a, cel7b, and cel3a) exhibited concomitant decrease in IR rates and increase in their gene expression in T. reesei under cellulase-producing condition (cellulose and lactose) that was accompanied with a more active NMD pathway, as compared to non cellulase-producing condition (glucose). In the presence of the NMD pathway inhibitor that successfully repressed the NMD pathway, the mRNA levels of cellulase genes were sharply down-regulated, but the rates of IR in these genes were significantly up-regulated. Consistently, the cellulase activities were severely inhibited. In addition, the NMD pathway inhibitor caused the downregulated mRNA levels of two important genes of the target of rapamycin (TOR) pathway, trfkbp12 and trTOR1. The absence of gene trfkbp12 made the cellulase production in T. reesei more sensitive to the NMD pathway inhibitor. Conclusion All these findings suggest that the IR of cellulase genes regulates their own gene expression by coupling with the NMD pathway, which might involve the TOR pathway. Our results provide better understanding on intron retention, the NMD pathway, and cellulase production mechanism in filamentous fungi.

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Decomposition of the metastable beta titanium alloy Ti-15-3

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100075117 ◽

1983 ◽

Vol 41 ◽

pp. 264-265

Author(s):

Y. L. Chen ◽

S. Fujlshiro

Keyword(s):

Low Cost ◽

High Strength ◽

Alpha Phase ◽

Thick Sheet ◽

Omega Phase ◽

Beta Titanium Alloy ◽

Beta Titanium ◽

Beta Matrix ◽

Metastable Beta ◽

Very High

Metastable beta titanium alloys have been known to have numerous advantages such as cold formability, high strength, good fracture resistance, deep hardenability, and cost effectiveness. Very high strength is obtainable by precipitation of the hexagonal alpha phase in a bcc beta matrix in these alloys. Precipitation hardening in the metastable beta alloys may also result from the formation of transition phases such as omega phase. Ti-15-3 (Ti-15V- 3Cr-3Al-3Sn) has been developed recently by TIMET and USAF for low cost sheet metal applications. The purpose of the present study was to examine the aging characteristics in this alloy.The composition of the as-received material is: 14.7 V, 3.14 Cr, 3.05 Al, 2.26 Sn, and 0.145 Fe. The beta transus temperature as determined by optical metallographic method was about 770°C. Specimen coupons were prepared from a mill-annealed 1.2 mm thick sheet, and solution treated at 827°C for 2 hr in argon, then water quenched. Aging was also done in argon at temperatures ranging from 316 to 616°C for various times.

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Chromic Acid Etching of Polyethylene

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100070801 ◽

1973 ◽

Vol 31 ◽

pp. 72-73

Author(s):

J. D. Muzzy ◽

R. D. Hester ◽

J. L. Hubbard

Keyword(s):

Chromic Acid ◽

Low Cost ◽

Acid Etching ◽

Crystalline Morphology ◽

Bulk Specimen ◽

Morphological Studies ◽

Perfect Form ◽

Molecular Length ◽

Lamellar Texture ◽

Cost Studies

Polyethylene is one of the most important plastics produced today because of its good physical properties, ease of fabrication and low cost. Studies to improve the properties of polyethylene are leading to an understanding of its crystalline morphology. Polyethylene crystallized by evaporation from dilute solutions consists of thin crystals called lamellae. The polyethylene molecules are parallel to the thickness of the lamellae and are folded since the thickness of the lamellae is much less than the molecular length. This lamellar texture persists in less perfect form in polyethylene crystallized from the melt.Morphological studies of melt crystallized polyethylene have been limited due to the difficulty of isolating the microstructure from the bulk specimen without destroying or deforming it.

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Glass Knife Geometry as Seen by the SEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100066322 ◽

1971 ◽

Vol 29 ◽

pp. 454-455

Author(s):

J. Temple Black

Keyword(s):

Low Cost ◽

Amorphous Material ◽

Stress Concentrations ◽

Basic Tool ◽

Tool Materials ◽

Optical Inspection ◽

Surface Flaws ◽

Slip Planes ◽

Glass Knife ◽

Diamond Knife

In ultramicrotomy, the two basic tool materials are glass and diamond. Glass because of its low cost and ease of manufacture of the knife itself is still widely used despite the superiority of diamond knives in many applications. Both kinds of knives produce plastic deformation in the microtomed section due to the nature of the cutting process and microscopic chips in the edge of the knife. Because glass has no well defined slip planes in its structure (it's an amorphous material), it is very strong and essentially never fails in compression. However, surface flaws produce stress concentrations which reduce the strength of glass to 10,000 to 20,000 psi from its theoretical or flaw free values of 1 to 2 million psi. While the microchips in the edge of the glass or diamond knife are generally too small to be observed in the SEM, the second common type of defect can be identified. This is the striations (also termed the check marks or feathers) which are always present over the entire edge of a glass knife regardless of whether or not they are visable under optical inspection. These steps in the cutting edge can be observed in the SEM by proper preparation of carefully broken knives and orientation of the knife, with respect to the scanning beam.

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Spectraplot: A PC-based spectrum translation and display program

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010010665x ◽

1988 ◽

Vol 46 ◽

pp. 918-919

Author(s):

H. O. Colijn

Keyword(s):

Low Cost ◽

Transfer Program ◽

File Transfer ◽

Transfer Data ◽

Error Checking ◽

Data Files ◽

The University

Many labs today wish to transfer data between their EDS systems and their existing PCs and minicomputers. Our lab has implemented SpectraPlot, a low- cost PC-based system to allow offline examination and plotting of spectra. We adopted this system in order to make more efficient use of our microscopes and EDS consoles, to provide hardcopy output for an older EDS system, and to allow students to access their data after leaving the university.As shown in Fig. 1, we have three EDS systems (one of which is located in another building) which can store data on 8 inch RT-11 floppy disks. We transfer data from these systems to a DEC MINC computer using “SneakerNet”, which consists of putting on a pair of sneakers and running down the hall. We then use the Hermit file transfer program to download the data files with error checking from the MINC to the PC.

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Transmission Electron Microscopy of oriented Co84Cr14Ta2 thin films for longitudinal magnetic recording

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100088099 ◽

1991 ◽

Vol 49 ◽

pp. 756-757

Author(s):

T. P. Nolan

Keyword(s):

Thin Film ◽

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Low Cost ◽

Low Noise ◽

Information Storage ◽

High Coercivity ◽

Magnetic Alloy ◽

Magnetic Media ◽

Transmission Electron

Thin film magnetic media are being used as low cost, high density forms of information storage. The development of this technology requires the study, at the sub-micron level, of morphological, crystallographic, and magnetic properties, throughout the depth of the deposited films. As the microstructure becomes increasingly fine, widi grain sizes approaching 100Å, the unique characterization capabilities of transmission electron microscopy (TEM) have become indispensable to the analysis of such thin film magnetic media.Films were deposited at 225°C, on two NiP plated Al substrates, one polished, and one circumferentially textured with a mean roughness of 55Å. Three layers, a 750Å chromium underlayer, a 600Å layer of magnetic alloy of composition Co84Cr14Ta2, and a 300Å amorphous carbon overcoat were then sputter deposited using a dc magnetron system at a power of 1kW, in a chamber evacuated below 10-6 torr and filled to 12μm Ar pressure. The textured medium is presently used in industry owing to its high coercivity, Hc, and relatively low noise. One important feature is that the coercivity in the circumferential read/write direction is significandy higher than that in the radial direction.

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Influence of MBE Growth Conditions on Dislocation Densities of GaAs/Ge Epilayers Grown on Silicon Substrates

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100118692 ◽

1985 ◽

Vol 43 ◽

pp. 364-365

Author(s):

K.M. Hones ◽

P. Sheldon ◽

B.G. Yacobi ◽

A. Mason

Keyword(s):

High Efficiency ◽

Lattice Mismatch ◽

Low Cost ◽

Growth Conditions ◽

Nonradiative Recombination ◽

Device Structure ◽

Si Substrates ◽

Transmission Electron ◽

Dislocation Densities ◽

Dislocation Type

There is increasing interest in growing epitaxial GaAs on Si substrates. Such a device structure would allow low-cost substrates to be used for high-efficiency cascade- junction solar cells. However, high-defect densities may result from the large lattice mismatch (∼4%) between the GaAs epilayer and the silicon substrate. These defects can act as nonradiative recombination centers that can degrade the optical and electrical properties of the epitaxially grown GaAs. For this reason, it is important to optimize epilayer growth conditions in order to minimize resulting dislocation densities. The purpose of this paper is to provide an indication of the quality of the epitaxially grown GaAs layers by using transmission electron microscopy (TEM) to examine dislocation type and density as a function of various growth conditions. In this study an intermediate Ge layer was used to avoid nucleation difficulties observed for GaAs growth directly on Si substrates. GaAs/Ge epilayers were grown by molecular beam epitaxy (MBE) on Si substrates in a manner similar to that described previously.

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A technique for protecting delicate specimens during processing

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100156894 ◽

1989 ◽

Vol 47 ◽

pp. 982-983

Author(s):

R.J. Mount ◽

R.V. Harrison

Keyword(s):

Basilar Membrane ◽

Low Cost ◽

Organ Of Corti ◽

Region Of Interest ◽

Sensory Epithelium ◽

Physical Damage ◽

Air Drying ◽

Specimen Handling ◽

Two Component

The sensory end organ of the ear, the organ of Corti, rests on a thin basilar membrane which lies between the bone of the central modiolus and the bony wall of the cochlea. In vivo, the organ of Corti is protected by the bony wall which totally surrounds it. In order to examine the sensory epithelium by scanning electron microscopy it is necessary to dissect away the protective bone and expose the region of interest (Fig. 1). This leaves the fragile organ of Corti susceptible to physical damage during subsequent handling. In our laboratory cochlear specimens, after dissection, are routinely prepared by the O-T- O-T-O technique, critical point dried and then lightly sputter coated with gold. This processing involves considerable specimen handling including several hours on a rotator during which the organ of Corti is at risk of being physically damaged. The following procedure uses low cost, readily available materials to hold the specimen during processing ,preventing physical damage while allowing an unhindered exchange of fluids.Following fixation, the cochlea is dehydrated to 70% ethanol then dissected under ethanol to prevent air drying. The holder is prepared by punching a hole in the flexible snap cap of a Wheaton vial with a paper hole punch. A small amount of two component epoxy putty is well mixed then pushed through the hole in the cap. The putty on the inner cap is formed into a “cup” to hold the specimen (Fig. 2), the putty on the outside is smoothed into a “button” to give good attachment even when the cap is flexed during handling (Fig. 3). The cap is submerged in the 70% ethanol, the bone at the base of the cochlea is seated into the cup and the sides of the cup squeezed with forceps to grip it (Fig.4). Several types of epoxy putty have been tried, most are either soluble in ethanol to some degree or do not set in ethanol. The only putty we find successful is “DUROtm MASTERMENDtm Epoxy Extra Strength Ribbon” (Loctite Corp., Cleveland, Ohio), this is a blue and yellow ribbon which is kneaded to form a green putty, it is available at many hardware stores.

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