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Rumen bacteria and application thereof

A technology of rumen bacteria and preservation number, which is applied in the field of microorganisms, can solve the problems of low caproic acid rate and low output, and achieve the effect of realizing waste recycling and recycling

Active Publication Date: 2017-06-13
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems of low production rate and low output of caproic acid-producing bacteria, the present invention provides a new strain of rumen bacteria (Clostridium sp.) CPB6, which was preserved in Guangdong Province on December 20, 2016. Microbial Culture Collection Center, preservation address: 5th Floor, Building 59, Guangdong Institute of Microbiology, No. 100 Xianlie Middle Road, Guangzhou City, preservation number: GDMCCNo: 60133

Method used

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  • Rumen bacteria and application thereof
  • Rumen bacteria and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Isolation, purification and cultivation of embodiment 1 bacterial classification

[0033] 1. Separation medium

[0034] Medium composition (100mL): 1g casein peptone, 0.25g yeast juice, 0.4g NaHCO 3 , 0.1g cysteine, 0.045g K 2 HPO4, 0.045g KH 2 PO 4 ,0.09g NaCl,0.009g MgSO 4 ·7H 2 O, 0.009g CaCl 2 , 0.1mg resazurin, 1mg hemin, 0.5mL lactic acid, 0.1ml trace element solution, 1ml vitamin solution. Distilled water 100ml. in:

[0035] Trace element solution (L): FeCl 2 4H 2 O 1.5g; CoCl 2 ·6H 2 O 0.19g; MnCl 2 4H 2 O 0.1g; ZnCl 2 0.07g; NaMO 4 2H 2 O 0.036g; NiCl 2 ·6H 2 O 0.024g; H 3 BO 3 0.006g; CuCl 2 2H 2 O0.002g; HCl (25% solution) 10ml.

[0036] Vitamin solution (L): p-aminobenzoic acid 0.04g; biotin 0.01g; niacin 0.1g; B 5 0.05g; B 6 0.15g; B 1 0.1g; B 12 0.05g; D.L-lipoic acid 0.03g; B 2 0.03g; folic acid 0.01g.

[0037] 2. Separation process

[0038] (1) Enrichment of strains

[0039] Caproic acid bacteria belong to Clostridium...

Embodiment 2

[0050] Characteristics and identification of embodiment 2 strains

[0051] 1. Morphological characteristics

[0052] (1) Colony morphology

[0053] The colonies are round, white raised, and sticky in texture.

[0054] (2) Cell morphology

[0055] Gram staining is positive, the bacteria are short rod-shaped, and the size of the bacteria is 2μm×0.5μm, such as figure 1 shown.

[0056] 2. Physiological and biochemical characteristics

[0057] Using Biolog AN MicroPlates TM The characteristics of the bacteria are shown in Table 1 and Table 2 through analysis with API 20A anaerobic identification system.

[0058] Table 1 Biolog characteristics of strain CPB6

[0059]

[0060]

[0061] Table 2 API characteristics of strain CPB6

[0062]

[0063] 3. Molecular biological characteristics

[0064] The whole genome of CPB6 was sequenced through the bacterial Pacbio platform, and the whole genome map of CPB6 was obtained. The length of its 16SrRNA gene is 1511bp, and the ...

Embodiment 3

[0070] The influence of embodiment 3 initial pH value on CPB6 producing hexanoic acid

[0071] From image 3 It can be seen that when the pH is 5-6.5, the hexanoic acid content of the strain increases significantly; and when the pH is greater than 7.5, almost no hexanoic acid is produced. When the pH is 5.5-6, the ability to produce acid is the strongest, and the pH is 5.5-6 is the best.

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Abstract

The invention relates to rumen bacteria and application thereof, and belongs to the technical field of microorganism. Aiming at the problem of low rate and low yield of hexanoic acid production of hexanoic acid production bacteria in the prior art, the invention provides a new strain of rumen bacteria (Clostridium sp.) CPB6 GDMCCNo:60133. The colonial morphology of the strain is a round and white bump, and thick in character. Scanning electron microscopy shows that the rumen bacteria is short-rod-shaped, the size of the rumen bacteria is 2 mu m * 0.5 mu m, the rumen bacteria is Gram staining positive bacteria, the rumen bacteria suitable growth temperature is 30-40 DEG C, pH is 5.0-6.5, and an acid and a gas can be produced by anaerobic growth for fermentation of glucose. The strain can use lactic acid for production of hexanoic acid, hexanoic acid production concentration is 16.6g / L, the highest yield can reach 5.29g / L / d; the strain also can be used for the production of the high added value of hexanoic acid from organic waste and wastewater containing the lactic acid, and waste resource recovery and recycling can be achieved.

Description

technical field [0001] The invention relates to a strain of rumen bacteria and its application, belonging to the technical field of microorganisms. Background technique [0002] Caproic acid is an important chemical raw material, which can be used in food spices, animal feed, green antibacterial agents, corrosion inhibitors and biofuels and other fields. In particular, caproic acid is also the precursor of ethyl caproate, the main aroma component of Luzhou-flavor liquor, and has broad application prospects. Clostridium klyuveri is the most researched and most in-depth microbial catalyzed production of caproic acid, which uses ethanol as electron donor and acetic acid as electron acceptor to synthesize caproic acid. Currently, this process is commonly used for the biosynthesis of hexanoic acid. [0003] In recent years, studies have shown that lactic acid can also be used as an electron donor for the synthesis of caproic acid, and the efficiency is even higher than that of ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/40C02F3/34C12R1/145C02F101/34
CPCC02F3/34C02F2101/34C12P7/40C12N1/205C12R2001/145
Inventor 陶勇朱晓宇何晓红李大平
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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