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A microbacterium pt3 with protein degradation function and its application

A microbacterium, PT3 technology, applied in the field of microorganisms, can solve the problems of cost reduction, low utilization rate of feed protein, inability to decompose and transform, etc.

Active Publication Date: 2021-03-12
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Researchers have improved the palatability of aquatic feed, added appropriate attractants, and adjusted the balance of amino acids in feed to increase the feed intake of farmed animals. However, in order to keep costs down, most of the protein in domestic feed is plant protein. , Aquaculture animals have low digestibility of these plant proteins, which cannot be completely decomposed and transformed by the digestive enzymes of aquatic animals, which is also the main limiting factor for the low utilization rate of feed protein

Method used

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  • A microbacterium pt3 with protein degradation function and its application
  • A microbacterium pt3 with protein degradation function and its application
  • A microbacterium pt3 with protein degradation function and its application

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a microbacterium PT3 with protein degradation function and its application, and its preservation number is: GDMCC No: 60923. The microbacterium PT3 of the present invention is isolated from the prawn intestine itself, and it has been verified that the bacterium can secrete protease extracellularly, hydrolyze the protein in the living environment, and after colonization in the intestine of aquaculture animals, it can assist it in the utilization of protein and decompose residues. The remaining protein in the bait is synthesized by its own bacterial protein for use by farmed animals, and then colonized the intestinal tract again to form a virtuous cycle and improve the utilization of feed protein. The gnotobiotic experiment between the bacteria and Artemia shows that the strain will not cause stress or poison to farmed animals at a higher concentration; the strain has a wide tolerance, and the salinity has little effect on the growth of bacteria, so it is very suitable for aquaculture. And it can function efficiently; it is of great significance in the mariculture industry, and the bacterium can be used as a good strain of new microecological preparations, and has a good application prospect.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to a microbacterium PT3 with protein degradation function and its application. Background technique [0002] With the development of the economy and the improvement of people's material living standards, the demand for aquatic products is increasing day by day. At present, my country is one of the important aquatic product production and export countries in the world. From January to November 2018 alone, the national aquatic product output has reached 53.7555 million tons. In order to increase the weight gain of farmed animals between units, the protein content in aquatic feed is always maintained at a high level, while the utilization rate of protein in feed for farmed animals is generally low, and the bait fed is only 25% feed protein It is absorbed and utilized by farmed animals, and the remaining 75% exists in the remaining bait or is discharged into the cultured water body w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P21/00A23K10/18A23K50/80C12R1/01
CPCA23K10/18A23K50/80C12N1/20C12P21/00C12N1/205C12R2001/01
Inventor 陈偿丁雄祺谢媚杨艺滢方建相管云雁云龙
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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