Microbacterium PT3 with protein degradation function and application thereof
A microbacterium, PT3 technology, applied in the field of microorganisms, can solve the problems of low digestibility of plant protein, inability to decompose and transform, low utilization rate of feed protein, etc.
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Embodiment 1
[0023] Example 1 Enrichment and screening of microbacterium PT3 that can secrete protease extracellularly
[0024] Sample source: Litopenaeus vannamei aquaculture water body in intelligent mariculture room of South China Sea Institute of Oceanology, Chinese Academy of Sciences.
[0025] The specific implementation steps are as follows: select healthy prawns with large vitality and full digestive tract in the prawn breeding pond, and bring them back to the laboratory immediately. The remaining seawater on the surface of the prawns was absorbed by filter paper, the body surface was wiped with 75% v / v alcohol for disinfection, dissected under aseptic conditions, and the complete intestinal tract was taken out. Rinse the intestinal tissue with sterile PBS (pH=7.4) buffer solution for 3 to 4 times, then put it into a 1.5mL sterile EP tube, add 1mL of PBS buffer solution to the sterile EP tube, and grind it evenly with a sterile grinding rod. Bacterial suspension was prepared by 10...
Embodiment 2
[0026] The identification of embodiment 2 microbacteria PT3 bacterial classification
[0027] 1. Morphological observation
[0028] The purely cultured PT3 strain was inoculated on the surface of 2216E solid medium by streaking method, and cultured in a 30°C incubator for 24 hours to observe the morphological characteristics of the colony. The colony is dot-shaped, opaque white, with smooth edges, and the colony is moist.
[0029] The 2216E solid medium is prepared by adding agar powder with a mass fraction of 1.5% to the 2216E liquid medium components, mixing and sterilizing. The 2216E liquid medium contains 5g of peptone per liter, 1g of yeast extract powder, 0.1g of ferric citrate, 19.45g of sodium chloride, 5.98g of magnesium chloride, 3.24g of sodium sulfate, 1.8g of calcium chloride, and 0.55g of potassium chloride , sodium carbonate 0.16g, potassium bromide 0.08g, strontium chloride 0.034g, boric acid 0.022g, sodium silicate 0.004g, sodium fluoride 0.0024g, ammonium n...
Embodiment 3
[0041] Example 3 Exploration of Optimal Growth Conditions for Microbacterium PT3
[0042] The effects of different pH, temperature and NaCl mass fraction (salinity) on the growth of Microbacterium PT3 were studied respectively. Among them, the different temperatures are: 20°C, 25°C, 30°C, 37°C, 43°C; the different pH values are: 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0; the different NaCl mass fractions are: 0 %, 1%, 2%, 3%, 4%, 5%.
[0043] In addition to the factors investigated in the above experiments, the fermentation conditions are to inoculate the microbacterium PT3 culture solution in the LB fermentation medium with an inoculation amount of 1% (v / v), and after cultivating at 30°C and 200rpm for 24 hours, take samples to measure the conditions OD 600 Value, the LB fermentation medium without inoculation was used as blank control. Take culture temperature, pH value, NaCl mass fraction as abscissa, OD 600 Values plot the respective curves on the ordinate. see results ...
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