High-performance feather degrading bacterium and application thereof

A feather and strain technology, applied in the field of microorganisms, can solve unseen problems and achieve the effects of high production efficiency, increased yield and simple production process

Active Publication Date: 2019-10-08
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There is no report in the prior art that can efficiently degrade feathers into high-purity proline

Method used

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  • High-performance feather degrading bacterium and application thereof
  • High-performance feather degrading bacterium and application thereof
  • High-performance feather degrading bacterium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: the screening of chryseobacterium X-Y4

[0063] (1) Sample source: Soil samples from pigeon farms where rotten feathers have been piled up for a long time.

[0064] (2) Strain screening:

[0065] The screening process includes the following steps:

[0066] a. Preparation of selection medium

[0067] b. Primary screening of feather-degrading bacteria

[0068] c. Re-screening of feather-degrading bacteria

[0069] The step a may further include: preparing feather solid culture medium and preparing feather liquid culture medium.

[0070] The step b may further include: from the air-dried pigeon plant soil sample, weigh 5g of soil and add it to 50ml of sterile water for enrichment culture, and cultivate it at 30°C and 160r / min for 12h; take the enrichment culture solution and press 10 -1 、10 -2 , 10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 Gradient dilution was spread on the feather solid medium, and when a single colony grew out, it was purified and...

Embodiment 2

[0077] Embodiment 2: strain identification.

[0078] 16sRNA sequence analysis method: PCR amplification uses bacterial 16sRNA universal primers:

[0079] 27F5'-AGAGTTTGATCMTGGCTCAG-3'

[0080] 1492R5'-GGTTACCTTGTTACGACTT-3'

[0081] dNTP (2 μL), primer F (2 μL), primer R (2 μL), Taq DNA polymerase (0.25 μL), deionized water (37.75 μL). Reaction conditions: pre-denaturation at 95°C for 10 minutes, deformation at 95°C for 30s, annealing at 55°C for 30s, extension at 72°C for 30s, a total of 25 cycles, and full extension at 72°C for 10 minutes.

[0082] Most of the 16S rDNA sequence was determined, as shown in SEQ ID No:1. The base sequences of the measured sequences were compared with BLAST on the NCBI website, and a phylogenetic tree based on the complete sequence of 16S rRNA was constructed. The results showed that the strain was 98% homologous to Chrysobacterium aureus. Therefore, it is determined that the present invention uses Flavobacterium tirrenicum, and the specifi...

Embodiment 3

[0083] Embodiment 3: the culture condition optimization of feather degrading strain

[0084] (1) bacterial strain: the bacterial strain Flavobacterium tirrenicum X-Y4 that embodiment 1 screens out

[0085] (2) Method steps:

[0086] A single factor test was used to investigate the effects of the strain's feather addition amount, shaker speed, initial pH, inoculation amount and fermentation temperature on the soluble protein yield.

[0087] a. Optimal test of feather addition amount

[0088] The fermentation conditions were 100mL / 250mL of feather liquid culture medium, 2% (v / v) inoculum size, 30°C, 160r / min, pH=7.0, 72h of shake flask fermentation, and the filling amount of feather was 0.5, 1.0g, 1.5g, 2.0g, 2.5g and 3.0g.

[0089] b. Optimization experiment of shaking table speed

[0090] The amount of feather added is based on the optimal result determined by the above test. Other fermentation conditions are 100mL / 250mL of feather liquid culture medium, 2% (v / v) inoculum,...

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Abstract

The invention discloses a high-performance feather degrading bacterium, having a classification name of Flavobacterium tirrenicum, has a strain number of X-Y4, and is collected at China Center for Type Culture Collection under CCTCC NO: M2019286 on April 23th, 2019. The invention also discloses application of Flavobacterium tirrenicum in the preparation of proline by degrading feather. The bacterium bred herein helps effectively achieve reuse of waste feather; the waste feather can be used as a liquid fertilizer for farmland irrigation; biomass resources are integrally utilized; environmentalpollution due to poultry farming is alleviated. The bacterium herein has high degrading capacity, the fermenting period is short, and a fermentation broth of the bacterium has proline content of 02% and above. The fermentation broth with rich proline can be prepared into liquid fertilizers which has the effects of promoting plant growth, relieving salt stress, relieving acid stress and the like, and the nutritional value and utilization rate of the fertilizers are increased.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a feather-degrading bacterium and its application. Background technique [0002] The mass consumption of animals produces a lot of feather waste. The traditional treatment methods are mainly landfill, incineration and gas production, which cause great harm to the environment. The research on the use of poultry feathers to prepare animal feed was earlier, and a certain scale of application development has been achieved, but 90% of the keratin in feathers has not been efficiently utilized, resulting in a waste of available resources. As the main component of feathers, keratin has been used in the cosmetics industry, or as a component of composite materials and fiber products. my country is extremely rich in keratin resources, especially in modern agriculture, large-scale poultry farming produces a large amount of keratin waste, of which feather waste is the most produ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/24C05F15/00C05F11/08C05F17/00C12R1/20
CPCC05F1/005C05F11/08C05F17/00C12P13/24C12N1/205C12R2001/20Y02W30/40Y02A40/20
Inventor 谢婧婧宋天顺李洋
Owner NANJING UNIV OF TECH
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