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Exiguobacterium sp. P6 for converting livestock and poultry breeding wastes and application thereof

A technology of livestock and poultry waste and microbacteria, which is applied in the direction of solid waste removal, application, and microbial-based methods, can solve problems such as difficult hydrolysis, and achieve excellent degradation effects, huge application potential, and excellent degradation ability.

Active Publication Date: 2022-05-27
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as a special kind of protein, feather keratin has a dense and complex structure and is rich in disulfide bonds, so it is difficult to be hydrolyzed by common proteases
The microbacteria disclosed in the above-mentioned published materials have a certain degradation effect on common protein substrates. For keratin with a dense and complex structure and rich in disulfide bonds, whether the above-mentioned strains still have the same effect and whether they can achieve ideal degradation effect, the above patent documents do not disclose
[0006] In the application field of agricultural microorganisms, Microbacterium aureus 8220605 (preservation number CGMCC No.14676, CN108102992 A) and Microbacterium acetyl AMCC 101217 (CGMCC No.16305, CN 110951656 A) have a significant effect on the control of tomato root-knot nematodes. In addition, there are no relevant literature disclosures on the utilization of microbacteria to realize resource conversion and utilization of low-value protein waste generated in the field of livestock and poultry breeding and processing

Method used

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  • Exiguobacterium sp. P6 for converting livestock and poultry breeding wastes and application thereof
  • Exiguobacterium sp. P6 for converting livestock and poultry breeding wastes and application thereof
  • Exiguobacterium sp. P6 for converting livestock and poultry breeding wastes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The inventors isolated and purified Exiguobacterium tumefaciens P6, as follows:

[0045] Exactobacter P6 was isolated and purified:

[0046] (1) Feather liquid medium (g / L): Feather 15, NaCl 0.5, K 2 HPO 4 1.0, KH 2 PO 4 0.4, MgCl 2 ·7H 2 O 0.1, CaCl 2 0.06, adjust pH to 7.5 with NaOH;

[0047] LB liquid medium (g / L): peptone 10, NaCl 10, yeast powder 10;

[0048] (2) Take 1 g of soil as seed inoculant, and cultivate it in 100 mL of feather liquid medium for 7 days; culture conditions: 25° C., 180 rpm;

[0049] (3) Take 100 μL of bacterial liquid to coat the LB plate, invert the plate in a 25°C incubator for 24 hours, and select the colonies with good growth on the plate to streak 3 consecutive times on the LB plate to purify the strain, and eliminate the growth in the passage process. Weak strains;

[0050] (4) Inoculate the purified bacterial strain in the feather liquid medium, cultivate at 25° C. at 180 rpm for 48 hours, observe the degree of feather de...

Embodiment 2

[0054] About strain identification of strain P6 in Example 1

[0055] (1) Morphological characteristics of strains

[0056] After culturing at 25°C for 48h on the LB plate, the colonies on the plate were observed to be light orange with a diameter of about 3mm, and the colonies were flat ( figure 2 A), Gram stain is positive ( figure 2 B). Bacteria were rod-shaped as observed by scanning electron microscopy, with a size of 0.71 μm × 1.36 to 2.21 μm ( figure 2 C).

[0057] (2) Phylogenetic analysis

[0058] A single colony of P6 strain was inoculated into liquid LB medium and cultured at 37°C overnight. Take 1 mL of the cultured bacterial solution and extract the total bacterial DNA according to the instructions of "Bacterial Genome Extraction Kit".

[0059] 16S universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-ACGGCTACCTTGTTACGACTT-3') were designed. Using bacterial genome as template, follow I-5 TM 2×High-Fidelity MasterMix (Beijing Qingke Biotechno...

Embodiment 3

[0064] The method for obtaining keratinase by fermentation of Exiguobacterium P6 is as follows:

[0065] In Example 1, in the process of separation, purification, fermentation and cultivation of Exiguobacterium tumefaciens P6, (4) after the cultivation, the medium residue was filtered off with gauze, centrifuged at 11,000 rpm for 15 min, and the supernatant of the fermentation broth was collected to be keratinase.

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Abstract

The invention belongs to the field of recycling and harmless utilization of agricultural and forestry wastes, particularly relates to Exiguobacterium sp. P6, and further relates to application of the Exiguobacterium sp. P6 in degradation of livestock and poultry breeding wastes and resource utilization of the wastes. The Exiguobacterium sp. P6 is preserved in the China General Microbiological Culture Collection Center (CGMCC) on March 01, 2022, the strain preservation number is CGMCC No.24453, and the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The invention mainly discloses an Exiguobacterium sp. P6 strain, application of the Exiguobacterium sp. P6 strain in degradation of livestock and poultry breeding wastes, a method for obtaining keratinase by fermentation culture of the strain, and application of the obtained keratinase in degradation of livestock and poultry breeding wastes. The Exiguobacterium sp. P6 provided by the invention can efficiently degrade wastes such as feathers and the like, the degradation rate of the feathers is up to 98%, and the Exiguobacterium sp. P6 has a wide application prospect in the aspects of livestock and poultry breeding wastes and waste resource utilization.

Description

technical field [0001] The invention belongs to the field of harmless utilization of agricultural and forestry waste resources, and particularly relates to an Exiguobacterium sp. P6, and also relates to the application of the above-mentioned Exiguobacterium P6 in degrading livestock and poultry breeding waste and recycling the waste. Background technique [0002] The protein content of poultry feathers exceeds 80%, and it is rich in essential amino acids and rich in macroelements, trace elements, vitamins and growth factors. It is considered to be a high-quality protein feed with unique value. The main component of poultry feathers, wool, animal fur, and hoof horns is keratin. Keratin is a hard protein with a dense and complex structure. It is difficult to be hydrolyzed by common proteases such as trypsin, pepsin and papain, and can only be hydrolyzed by proteases called "keratinase". Therefore, keratinase plays an important role in the degradation and recycling of keratin....

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/52A23K10/12A23K10/26B09B3/60C12R1/01
CPCC12N1/20C12N9/52A23K10/12A23K10/26B09B3/00Y02P60/87
Inventor 边斐张燕岳寿松姚强宋恩亮王艳芹姜富贵成海建马德源
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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