CN104099273A - Culture medium for specifically separating intestinal bacteria capable of decomposing TMAO (tirmethylamine N-oxide) as well as preparation method and application thereof - Google Patents

Culture medium for specifically separating intestinal bacteria capable of decomposing TMAO (tirmethylamine N-oxide) as well as preparation method and application thereof Download PDF

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CN104099273A
CN104099273A CN201410330857.XA CN201410330857A CN104099273A CN 104099273 A CN104099273 A CN 104099273A CN 201410330857 A CN201410330857 A CN 201410330857A CN 104099273 A CN104099273 A CN 104099273A
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tmao
intestinal bacteria
substratum
decomposing
decomposed
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CN104099273B (en
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杨金龙
殷素会
许国洋
薛梅
杨晶旭
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Chongqing weighing detection Certification Technology Co., Ltd.
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Chongqing Academy of Animal Sciences
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor

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Abstract

The invention discloses a culture medium for specifically separating intestinal bacteria capable of decomposing TMAO (tirmethylamine N-oxide) as well as a preparation method and an application thereof. The culture medium comprises components as follows: 0.1-2.0 g/L of K2PO4, 0.1-2.0 g/L of yeast extract powder, 0.1-2.0 g/L of TMAO and a solvent of water, and the pH is 7.1-7.5. The components of the culture medium are simple and low in cost, and TMAO is taken as the sole carbon source, so that development of specifically separated bacteria capable of decomposing TMAO is better facilitated, and the culture medium can be applied to specific separation of the intestinal bacteria capable of decomposing TMAO.

Description

Specific isolation is decomposed intestinal bacteria substratum of trimethylamine oxide and its preparation method and application
Technical field
The invention belongs to microorganism field, be specifically related to the intestinal bacteria substratum that specific isolation is decomposed trimethylamine oxide, also relate to the preparation method and application of this substratum.
Background technology
Trimethylamine oxide (Tirmethylamine N-oxide, TMAO) has special delicate flavour, is a kind of novel animal phagostimulant, and its chemical structure is similar to methyl donor choline, trimethyl-glycine and S-adenosylmethionine etc.As intermediate metabolites important in animal body, there is important biochemical functions at aspects such as stabilizing protein structure, osmoregulation, anti-ionic unstabilities, in animal cultivation, be widely applied, but the increase of TMAO content in animal body, the cholesterol that can make to remove on vessel wall becomes more difficult, thereby the cardiovascular disordeies such as arteriosclerosis occur.
Flora in enteron aisle has a significant impact the accumulation of TMAO.In mouse animal experiment, supplementing TMAO lead can increase TMAO accumulation, also makes cardiovascular disorder probability double, and after killing normal the gut flora with microbiotic, supplementary TMAO lead does not affect cardiovascular disorder probability.And vegetarian and predator's intestinal microflora are widely different, so the utilization of TMAO is just had to difference.In addition, the intestinal microflora of these " food meat " is can be derivative, after having eaten for some time meat, these bacteriums are induced out, produce the ability of TMAO with regard to grow, therefore, be necessary that the intestinal bacteria to decomposing TMAO carries out separation and Culture, the nutritional condition that decomposes the growth of TMAO entero-bacte is studied.And the TMAO intestinal bacteria of decomposing being separated to is added in food after breeding, thereby reach the object that the cardiovascular disordeies such as prevention of arterial sclerosis occur.
The substratum of the intestinal bacteria of TMAO is decomposed in existing cultivation, the concentration of its composition and each composition is as follows: peptone 10g/L, beef extract powder 10g/L, sodium-chlor 5g/L, yeast soaks powder 1g/L, trimethylamine oxide 1g/L, agar 15g/L, but the intestinal bacteria that this substratum separation and Culture obtains is not because being taking trimethylamine oxide as sole carbon source, in the time of separation of bacterial, not only can be separated to the bacterium of decomposing trimethylamine oxide, can also be separated to the bacterium of decomposition of protein peptone simultaneously, its specificity is poor, and cost is high.Therefore, be badly in need of a kind of culture effect good, the specificity that cost is low is decomposed TMAO intestinal bacteria substratum.
Summary of the invention
In view of this, one of object of the present invention is to provide specific isolation to decompose the intestinal bacteria substratum of trimethylamine oxide, and scientific formulation is reasonable, is conducive to the growth that specificity is decomposed the intestinal bacteria of trimethylamine oxide; Two of object of the present invention is to provide specific isolation to decompose the preparation method of the intestinal bacteria substratum of trimethylamine oxide; Three of object of the present invention is to provide specific isolation to decompose the application of the intestinal bacteria substratum of trimethylamine oxide.
For achieving the above object, the invention provides following technical scheme:
1, specific isolation is decomposed the intestinal bacteria substratum of trimethylamine oxide, comprises following component: K 2pO 40.1-2.0g/L, yeast soaks powder 0.1-2.0g/L, TMAO0.1-2.0g/L, solvent is water, pH value is 7.1～7.5.
This substratum is taking TMAO as sole carbon source, does not contain peptone.
Preferably, also comprise agar 10-20g/L.
Preferred, comprise following component: K 2pO 40.2g/L, yeast soaks powder 0.6g/L, TMAO0.6g/L, solvent is water, pH value is 7.1.
Preferred, comprise following component: K 2pO 40.1g/L, yeast soaks powder 0.5g/L, TMAO0.5g/L, solvent is water, pH value is 7.2.
Preferred, comprise following component: K 2pO 41g/L, yeast soaks powder 1g/L, TMAO0.8g/L, solvent is water, pH value is 7.3.
Preferred, comprise following component: K 2pO 40.8g/L, yeast soaks powder 0.9g/L, TMAO0.9g/L, solvent is water, pH value is 7.5.
2, described specific isolation is decomposed the preparation method of the intestinal bacteria substratum of trimethylamine oxide, comprise the steps, by proportioning take K2PO4, yeast soaks powder and TMAO is dissolved in the water of proportional quantity, mixes, and regulates pH value to 7.3 ± 0.2.
3, the intestinal bacteria substratum that described specific isolation is decomposed trimethylamine oxide is in the application separating in the intestinal bacteria of decomposing trimethylamine oxide.
Beneficial effect of the present invention is: the invention discloses specific isolation and decompose intestinal bacteria substratum of trimethylamine oxide and its preparation method and application, this substratum can ensure under the condition of the good growth of intestinal bacteria of decomposing trimethylamine oxide (TMAO), can greatly reduce costs, taking trimethylamine oxide as sole carbon source, more scientific reasonable, decompose the intestinal bacteria of oxidation TMAO for specificity and created suitable growing environment, compared with commercially available TMAO substratum, not only cost 20%-30%, and the specificity growing decomposition trimethylamine oxide thalline number has improved 50%-500%, can be widely used in specific isolation and decompose the intestinal bacteria of TMAO.
Brief description of the drawings
In order to make object of the present invention, technical scheme and beneficial effect clearer, the invention provides following accompanying drawing:
Fig. 1 is that embodiment 5 separates the intestinal bacteria result figure that decomposes TMAO.
Fig. 2 is that comparative example 1 separates the intestinal bacteria result figure that decomposes TMAO.
Fig. 3 is that embodiment 6 separates the intestinal bacteria result figure that decomposes TMAO.
Fig. 4 is that comparative example 1 separates the intestinal bacteria result figure that decomposes TMAO.
Embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.The experimental technique of unreceipted actual conditions in embodiment, the condition of conventionally advising according to normal condition or according to manufacturer.
Embodiment 1
Specific isolation is decomposed the intestinal bacteria substratum of TMAO, and its formula is as follows: K 2pO 40.2g/L, yeast soaks powder 0.6g/L, TMAO0.6g/L, solvent is water.
The preparation method that specific isolation is decomposed the intestinal bacteria substratum of TMAO, gets 0.2g K 2pO 4, 0.6g yeast soaks powder and 0.6g TMAO is dissolved in 1L distilled water, mixes, and regulating pH value is 7.1.
Embodiment 2
Specific isolation is decomposed the intestinal bacteria substratum of TMAO, and its formula is as follows: K 2pO 40.1g/L, yeast soaks powder 0.5g/L, TMAO0.5g/L, solvent is water.
The preparation method that specific isolation is decomposed the intestinal bacteria substratum of TMAO, gets 0.1g K 2pO 4, 0.5g yeast soaks powder and 0.5g TMAO is dissolved in 1L distilled water, mixes, and regulating pH value is 7.2.
Embodiment 3
Specific isolation is decomposed the intestinal bacteria substratum of TMAO, and its formula is as follows: K 2pO 41g/L, yeast soaks powder 1g/L, TMAO0.8g/L, solvent is water.
The preparation method that specific isolation is decomposed the intestinal bacteria substratum of TMAO, gets 1g K 2pO 4, 1g yeast soaks powder and 0.8g TMAO is dissolved in 1L distilled water, mixes, and regulating pH value is 7.3.
Embodiment 4
Specific isolation is decomposed the intestinal bacteria substratum of TMAO, and its formula is as follows: K 2pO 40.8g/L, yeast soaks powder 0.9g/L, TMAO0.9g/L, solvent is water.
The preparation method that specific isolation is decomposed the intestinal bacteria substratum of TMAO, gets 0.8g K 2pO 4, 0.9g yeast soaks powder and 0.9g TMAO is dissolved in 1L distilled water, mixes, and regulating pH value is 7.5.
The substratum of above-described embodiment 1～4 is the composition of liquid nutrient medium, as need preparation solid medium adds 10-20g agar in substratum, uses front at 121 DEG C of autoclaving 20min.
Embodiment 5
The intestinal bacteria substratum that utilizes the specific isolation making in embodiment 2 to decompose TMAO separates the intestinal bacteria of decomposing TMAO, and concrete steps are as follows:
Get after adding stroke-physiological saline solution 10mL in intestine of young pigs faecal samples 1g and place 30 minutes in room temperature (18-25 DEG C) is lower after sufficient concussion, abandon throw out, getting 1mL supernatant liquid is inoculated in the separation that embodiment 2 obtains and decomposes in the intestinal bacteria solid medium of TMAO, under 30-37 DEG C of condition, cultivate 1 day, result as shown in Figure 1.
Comparative example 1
Utilize commercially available trimethylamine oxide substratum, each component and concentration are as follows: peptone 10g/L, beef extract powder 10g/L, sodium-chlor 5g/L, yeast soaks powder 1g/L, trimethylamine oxide 1g/L and agar 15g/L, separate the intestinal bacteria of decomposing TMAO according to the method for embodiment 5, result as shown in Figure 2.
Result shows, the colony number that the intestinal bacteria substratum that utilizes separation that embodiment 2 makes to decompose TMAO grows is higher than the height of commercially available substratum, high approximately 5%, show that separation that the present invention obtains decomposes the growth that the intestinal bacteria substratum of TMAO is conducive to the intestinal bacteria of decomposing TMAO, and cost is lower, low 20%-30%.
Embodiment 6
The intestinal bacteria substratum that utilizes the separation making in embodiment 1 to decompose TMAO separates the intestinal bacteria of decomposing TMAO, and the present embodiment uses solid medium, and concrete grammar is as follows:
Get and in intestine of young pigs faecal samples 1g, add after stroke-physiological saline solution 10mL room temperature after sufficient concussion to place 30 minutes, abandon throw out, on the intestinal bacteria substratum of the separation decomposition TMAO that liquid inoculation is obtained in embodiment 1 (solid), carry out four ride cultivations, cultivate 2 days at 20-30 DEG C, result as shown in Figure 3.
Comparative example 2
Utilize commercially available trimethylamine oxide substratum, each component and concentration are as follows: peptone 10g/L, beef extract powder 10g/L, sodium-chlor 5g/L, yeast soaks powder 1g/L, trimethylamine oxide 1g/L and agar 15g/L, separate the intestinal bacteria of decomposing TMAO according to the method for embodiment 5, result as shown in Figure 4.
Result shows, the colony number that the intestinal bacteria substratum that utilizes separation that embodiment 1 makes to decompose TMAO grows is higher than commercially available substratum, high approximately 10%, show that separation that the present invention obtains decomposes the growth that the intestinal bacteria substratum of TMAO is conducive to the intestinal bacteria of decomposing TMAO, and cost is lower, low 20%-30%.
Embodiment 3 and 4 makes and separates that to decompose the intestinal bacteria substratum of TMAO similar with 2 to embodiment 1, and specificity colony number is respectively higher than 8% and 12% of commercially available substratum.
Hence one can see that, and the intestinal bacteria substratum that TMAO is decomposed in the separation that the embodiment of the present invention 1～4 makes is the substratum of a high efficiency, low cost, is more conducive to decompose the growth of TMAO intestinal bacteria.
Finally explanation is, above preferred embodiment is only unrestricted in order to technical scheme of the present invention to be described, although the present invention is described in detail by above preferred embodiment, but those skilled in the art are to be understood that, can make various changes to it in the form and details, and not depart from the claims in the present invention book limited range.

Claims (8)

1. specific isolation is decomposed the intestinal bacteria substratum of trimethylamine oxide, it is characterized in that, comprises following component: K 2pO 40.1-2.0g/L, yeast soaks powder 0.1-2.0g/L, TMAO0.1-2.0g/L, solvent is water, pH value is 7.1～7.5.
2. specific isolation is decomposed the intestinal bacteria substratum of trimethylamine oxide according to claim 1, it is characterized in that: also comprise agar 10-20g/L.
3. specific isolation is decomposed the intestinal bacteria substratum of trimethylamine oxide according to claim 1, it is characterized in that, comprises following component: K 2pO 40.2g/L, yeast soaks powder 0.6g/L, TMAO0.6g/L, solvent is water, pH value is 7.1.
4. specific isolation is decomposed the intestinal bacteria substratum of trimethylamine oxide according to claim 1, it is characterized in that, comprises following component: K 2pO 40.1g/L, yeast soaks powder 0.5g/L, TMAO0.5g/L, solvent is water, pH value is 7.2.
5. specific isolation is decomposed the intestinal bacteria substratum of trimethylamine oxide according to claim 1, it is characterized in that, comprises following component: K 2pO 41g/L, yeast soaks powder 1g/L, TMAO0.8g/L, solvent is water, pH value is 7.3.
6. specific isolation is decomposed the intestinal bacteria substratum of trimethylamine oxide according to claim 1, it is characterized in that, comprises following component: K 2pO 40.8g/L, yeast soaks powder 0.9g/L, TMAO0.9g/L, solvent is water, pH value is 7.5.
7. the preparation method that described in claim 1-6 any one, specific isolation is decomposed the intestinal bacteria substratum of trimethylamine oxide, is characterized in that: comprise the steps, get K by proportioning 2pO 4, yeast soaks powder and TMAO, is then dissolved in the water of proportional quantity, mixes, and regulates pH value to 7.1～7.5.
8. the application of the intestinal bacteria substratum of specific isolation decomposition trimethylamine oxide in the intestinal bacteria of specific isolation decomposition trimethylamine oxide described in claim 1-6 any one.
CN201410330857.XA 2014-07-11 2014-07-11 Specific isolation decomposes intestinal bacteria culture medium of trimethyloxamine and its preparation method and application Active CN104099273B (en)

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CN201410330857.XA CN104099273B (en) 2014-07-11 2014-07-11 Specific isolation decomposes intestinal bacteria culture medium of trimethyloxamine and its preparation method and application
PCT/CN2014/083520 WO2016004658A1 (en) 2014-07-11 2014-08-01 Culture medium for specifically separating intestinal bacteria capable of decomposing trimethylamine n-oxide as well as preparation method therefor and application thereof

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107325993A (en) * 2017-08-31 2017-11-07 重庆市畜牧科学院 High-efficient culture chicken rhinitis haemophilus paragallinarum culture medium and its preparation method and application

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WO2000029604A1 (en) * 1998-11-18 2000-05-25 The University Of Akron Production of biological materials by simultaneous aerobic and anaerobic respiration
CN103734530A (en) * 2013-12-31 2014-04-23 佛山市信豚生物科技有限公司 Feed inducing agent for aquatic feed

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WO2000029604A1 (en) * 1998-11-18 2000-05-25 The University Of Akron Production of biological materials by simultaneous aerobic and anaerobic respiration
CN103734530A (en) * 2013-12-31 2014-04-23 佛山市信豚生物科技有限公司 Feed inducing agent for aquatic feed

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张晓静: "腐败希瓦氏菌代谢氧化二甲胺相关研究", 《中国优秀硕士学位论文全文数据库》, 15 February 2013 (2013-02-15), pages 12 *
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107325993A (en) * 2017-08-31 2017-11-07 重庆市畜牧科学院 High-efficient culture chicken rhinitis haemophilus paragallinarum culture medium and its preparation method and application

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Effective date of registration: 20170818

Address after: 402460, room 4, building 53, 301 Chang Long Avenue, Chang Zhou street, Rongchang District, Chongqing

Patentee after: Chongqing weighing detection Certification Technology Co., Ltd.

Address before: 402460 Rongchang County State Road, Chongqing, No. 770

Co-patentee before: Yang Jingxu

Patentee before: Chongqing Academy of Animal Sciences