Enzymes from conidiobolus brefeldianus and process for preparation thereof
A kind of ear mold, selected technology, applied in the field of enzyme and its preparation from Burfred ear mold (CONIDIOBOLUS BREFELDIANUS), can solve the problem of no inclusion etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0082] This example illustrates the isolation of O. bulfredii MTCC 5185 and its characterization by morphological characteristics. Isolation of fungal cultures from decomposing plant debris in Pune (collection), Maharashtra, India. Spread fine particles of plant debris on MGYP agar (malt extract-0.3%; yeast extract-0.3%; peptone-0.5, glucose-1%, agar-2%) attached to the inner surface of the petri dish lid Block and incubate the plate at 28°C. Single isolated colonies arising from the vigorously irradiated conidia were picked and transferred to MGYP agar slants and incubated at 28°C for 2-3 days. The organism grows rapidly and is identified as belonging to the genus Atomyces based on the morphology of strongly emanating large spherical conidia with basal papillae. The mycelia are non-septate and multinucleate, but become septate in later stages. Conidiophores have short filaments and are indistinguishable from mycelia. At the tip of the molecular spore peduncle, strongly ra...
Embodiment 2A
[0084] This example illustrates the isolation of genomic DNA from O. bulfredii MTCC 5185 by the cetyltrimethylammonium bromide (CTAB) method. To isolate DNA, the fungus was grown in liquid flasks in malt extract dextrose yeast extract peptone (MGYP) medium consisting of (g / L) malt extract-3; yeast extract -3; peptone-5 and glucose-10. Growth was initiated by inoculating spores from 7 day old MGYP slants. The flasks were incubated for 48 hours at 28°C on a rotary shaker (200 rpm). The contents were centrifuged at 8000rpm for 15min, and washed repeatedly to remove medium components. 3-5 g of wet mycelium was ground in liquid nitrogen, then 8-10 ml of CTAB extraction buffer containing 0.2% β-mercaptoethanol, pH 8 was added, followed by 20 μl of proteinase K (20 mg / ml) and incubated at 65°C 1h. This was followed by the addition of 20 μl RNase A (10 mg / ml) and further incubation at 65° C. for 15 min. To the supernatant collected after centrifugation (8000 rpm, 10 min), 10 ml o...
Embodiment 2B
[0086] This example illustrates genomic DNA polymerase chain reaction (PCR) amplification for the 18S rDNA gene. The primers used to identify fungal species were universal fungal 18S ribosomal DNA (rDNA) primers NS1-F (GTA GTC ATA TGC TTG TCT C), NS8-R (TCC GCA GGT TCA CCT ACG GA). Polymerase chain reaction (25 μl) was used to amplify the 18S rDNA gene from genomic DNA. The reaction mixture typically contains genomic DNA - 0.70 μl, 10X PCR buffer - 2.50 μl, 0.2 mM dNTPs - 2.5 μl, forward and reverse primers each 10-20 pmol - 1.25 μl, distilled water - 16.60 μl and 1 unit of Taq DNA Polymerase - 0.20 μl. PCR conditions for 18S rDNA gene propagation were: initial denaturation at -95°C for 3 min, followed by 35 cycles of 94°C for 1 min, 57°C for 30 sec, 72°C for 2 min, and a final extension at 72°C for 10 min. 5 μl of the above PCR amplification product was used to check the amplification on a 1.0% agarose gel.
PUM
| Property | Measurement | Unit |
|---|---|---|
| molecular weight | aaaaa | aaaaa |
| volume | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More