Myceliophthora thermophila as well as solid-state fermentation method and application thereof
A thermophilic Myceliophthora thermophila and solid-state fermentation technology, applied in the field of microorganisms, can solve the problems of difficult rapid and effective degradation, long decay period, unbalanced nutrients, etc. Effect
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Embodiment 1
[0029] Screening and identification of Myceliophthora thermophila:
[0030] 1) Dilute the obtained soil sample with sterile water, then spread it on a PDA plate containing penicillin, and after culturing at 40°C for 3 days, select mold colonies and transfer them to the PDA plate to obtain a single mold colony.
[0031] The total DNA of the mold was extracted by the CTAB method, and then amplified by PCR with fungal universal primers ITS1 and ITS4. The product was detected by 1% agarose gel electrophoresis and then sequenced. The sequencing results were analyzed and compared on NCBI, and the strain was determined to be thermophilic myceliophthora. mold.
[0032] 2) Mutagenesis and breeding of high-yield xylanase strain Myceliophthora thermophila MT1810.
[0033] Fermentation medium: Miscanthus 5.0g and nutrient solution 10ml, sterilized at 121°C for 30min.
[0034] Nutrient solution: (NH 4 ) 2 SO 4 3.5g, KH 2 PO 4 3.0g, MgSO 4 ·7H 2 O0.5g, CaCl 2 0.5g, Tween80 10.0g a...
Embodiment 2
[0057] Solid-state fermentation method and enzymatic properties of Myceliophthora thermophila MT1810 with high xylanase production:
[0058] Solid-state fermentation medium: 70 parts of wheat bran, 3 parts of yeast extract, 10 parts of ammonium sulfate, 7 parts of potassium dihydrogen phosphate, 10 parts of glucose, the ratio of material to water is 1:1.5, and the initial pH is 7.5-8.0.
[0059] Every 100g of the above-mentioned solid-state fermentation medium was inoculated with 2ml of the spore suspension (the number of spores was 5×10 7 pcs / ml), stirred evenly, 30 ℃ static culture for 3 days, 50 ℃ cold air drying to moisture 12%, recorded xylanase enzyme activity 1860U / g, the number of viable bacteria 2.3 billion CFU / g.
[0060] Prepare Myceliophthora thermophila MT1810 spore suspension: the culture medium is YPD plate or YPD slant bottle, after inoculation with Myceliophthora thermophila MT1810, culture aseptically at 40±3°C for 4-6 days, then use sterile water (> 15ml, C...
Embodiment 3
[0072] Application of Myceliophthora thermophila MT1810 in the preparation of decomposing agent:
[0073] For the decomposing experiment of rice straw, 2kg of rice straw was cut to less than 3cm, and the ingredients of rice straw used were: 32.56% cellulose, 18.23% hemicellulose, 15.03% lignin, 3.56% crude protein, 1.38% crude fat, and 13.24% ash . Add 3% urea by weight of straw, 1% potassium dihydrogen phosphate, and adjust the moisture content to 70%. The spore suspension of Myceliophthora thermophila MT1810 was inoculated in 1kg rice straw powder, the inoculum size was 5×10 5 straw per g; 1 kg grass powder not inoculated with Myceliophthora thermophila was used as the control group. Afterwards, the grass powder was put into nylon mesh bags and placed in a 40°C incubator to decompose, and there was no need to turn the pile during this period.
[0074] After 30 days, the decomposing experiment was finished. After inoculation with Myceliophthora thermophila and decomposing,...
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