CN106148230B - The application of one plant of false chainlet Bifidobacterium and its preparation conjugated linoleic acid or conjugate linolenic acid - Google Patents
The application of one plant of false chainlet Bifidobacterium and its preparation conjugated linoleic acid or conjugate linolenic acid Download PDFInfo
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Abstract
The present invention relates to one plant of false chainlet Bifidobacterium CCFM748 and application thereof.Free linoleic acid, linolenic acid efficiently can be separately converted to the more conjugated linoleic acid of bioactivity, conjugate linolenic acid by present invention vacation chainlet Bifidobacterium CCFM748, it can be directly used for preparation conjugated linoleic acid, conjugate linolenic acid, it can also be used to food of the production rich in conjugated linoleic acid, conjugate linolenic acid.
Description
[technical field]
The present invention relates to the false chainlet Bifidobacterium (Bifidobacterium that one plant is isolated from Neonatal Faeces
Pseudocatenulatum) the application of CCFM748 and the bacterial strain in preparation conjugated linoleic acid or conjugate linolenic acid.
[background technique]
Conjugated linoleic acid (Conjugated linoleic acid, CLA) is a kind of 18 carbon two containing conjugated double bond
Olefin(e) acid general name is the position isomer and geometric isomer of linoleic acid (Linoleic acid, 18:2).It is most abundant, most common
Isomers is suitable 9, anti-11-CLA (c9, t11-CLA), also referred to as rumenic acid (Rumenic acid).In addition, anti-10, along 12-
CLA (t10, c12-CLA) is also the relatively high isomers of nature content.Conjugated linoleic acid is closed because of its biological function
Note, physiological activity specifically include that anticancer, it is anti-inflammatory, slow down atherosclerosis, weight-reducing, alleviate diabetes and promote bone raw
Long and immunological regulation etc..Different isomers has a different physiological functions, wherein c9, t11-CLA and t10, c12-CLA be by
The conjugated linoleic acid isomers of generally acknowledged most physiological activity, the most important function of c9, t11-CLA are anticancer, anti-inflammatory and exempt from
Epidemic disease adjusting etc., and t10, c12-CLA are very significant for the influence in terms of weight-reducing and lipid-metabolism.In addition, t9,
T11-CLA is also reported with anti-inflammatory isoreactivity.
Conjugate linolenic acid (Conjugated linolenic acid, CLNA) be by linolenic acid (Linolenic acid,
LNA it) is derived the general name of octatecatrienoic acid a variety of positions and geometric isomer with conjugated double bond, it has a variety of battalion
It supports and healthcare function, such as anticancer anti-diabetic, antiatherosclerosis, reduces body fat content, insulin resistance, adjusts body
A variety of nutrition and health care functions such as immune, it has also become the research hotspot in the fields such as medicine, chemistry, nutrition.In conjugate linolenic acid
Various stereoisomers in, c9, t11, c15-CLNA (CLNA1), t9, t11, c15-CLNA (CLNA2), t10, c12, c15-
CLNA and c6, c9, t11-CLNA etc. are considered as the isomers of most bioactivity.
Natural conjugated linoleic acid is primarily present in the butterfat and meat products of cud animal ox, sheep etc., in every gram of butterfat
Content is from 2mg-25mg etc., and CLA content increases with the milk cow age and increased.The non-natural origin of CLA mainly passes through manually
Synthetic method obtains, and then because of its raw material and method difference, the content of Isomers differs greatly artificial synthesized CLA.It is natural
Some vegetable seeds pomegranate seeds, bancoul nuts, bitter melon seed, pot marigold seed, snakegourd and the blue flower principal columns of a hall seed on boundary etc. are sub- rich in conjugation
Numb acid.Only Seeds of Trichosanthes kirilowii is direct-edible in numerous vegetable seeds containing conjugate linolenic acid, and the grease in plant seed at
It is point very complicated, realize that the isolation and purification of conjugate linolenic acid is very difficult.
In addition, the prior art also can produce conjugate linolenic acid, but this method yield by alkali process linolenic acid isomerization process
It is lower, and there is reagent residual to be difficult to put into practical application in product, therefore the industrialized production of conjugate linolenic acid is not yet realized so far.
At present studies have found that by microorganism conversion CLA, CLNA, some special lactic acid bacterias are with conjugated linoleic acid and altogether
Conjugated linolenic conversion capability, such as Gorissen etc. carry out more than 30 plants of Bifidobacterium bioconversion CLA and conjugate linolenic acid
Research, CLA or CLNA can be produced by finding 6 plants in 36 plants of Bifidobacteriums, be turned in 6 plants of Bifidobacteriums to two kinds of conjugated fatty acids
Highest rate is respectively 53%, 78% (Gorissen L, et al.Production of conjugated linoleic
acid and conjugatedlinolenic acid isomers by Bifidobacterium species[J].Appl
Microbiol Biotechnol,2010,87(6):2257-2266.).However, gained is recorded according to it, gained fatty acid
Isomers is not with c9, t11-CLA and the t10 of most bioactivity, c12-CLA or c9, t11, c15-CLNA and t9, t11,
Based on c15-CLNA.
[summary of the invention]
The purpose of the present invention is overcoming prior art defect, acquisition conjugated linoleic acid, conjugate linolenic acid yield are higher, convert
Rate is higher and product in c9, t11-CLA and t10, c12-CLA or c9, t11, c15-CLNA and t9, t11, c15-CLNA be
The false chainlet Bifidobacterium of main isomer.
The present invention also provides application of the above-mentioned bacterial strains in preparation conjugated linoleic acid or conjugate linolenic acid.
To achieve the goals above, the present invention provides one plant of vacation chainlet Bifidobacterium CCFM748 bacterial strain in 2016 6
The moon 12 is in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, deposit number CGMCC
No.12603。
False chainlet Bifidobacterium CCFM748 of the invention is new separation, screening acquisition, extracted by bacterial genomes DNA,
The amplification of 16S rDNA specific primer PCR, amplified production purifying, DNA sequencing, sequence alignment and etc. bacterial 16 S rDNA sequence survey
Sequence method carries out Species estimation, is accredited as false chainlet Bifidobacterium, and be named as false chainlet Bifidobacterium CCFM748.
False chainlet Bifidobacterium CCFM748 of the invention has following biological characteristics:
Thallus feature: thallus is creamy white.
Colony characteristics: the bacterium colony protrusion on mMRS solid plate, smooth, round, milky, translucent, diameter be 1~
2mm。
Growth characteristics: under conditions of 37 DEG C of constant-temperatureanaerobic anaerobics, false chainlet Bifidobacterium CCFM748 is trained in mMRS culture medium
It supports and about reaches late log phase for 24 hours.
False chainlet Bifidobacterium is one of Bifidobacterium.Bifidobacterium includes 35 kinds altogether, including the youth is double
Discrimination bacillus, animal bifidobacteria animal subspecies, bifidobacterium animalis acid subspecies (i.e. bifidobacterium lactis), bifidobacterium bifidum,
Bear peak Bifidobacterium, cloth nurse Bifidobacterium, bifidobacterium breve, chain Bifidobacterium, globefish Bifidobacterium, bifidobacterium coryneforme, rabbit
The long subspecies of Bifidobacterium, bifidobacterium dentium, bifidobacterium gallicum, Bifidobacterium gallinarum, bifidobacterium longum, bifidobacterium longum baby
Subspecies, big Bifidobacterium, Bifidobacterium minimum, false chainlet Bifidobacterium, the false long subspecies of bifidobacterium pseudolongum, bifidobacterium pseudolongum
Ball revolves worm subspecies, Bifidobacterium pullorum, bifidobacterium subtile, bifidobacterium thermophilum etc..
Since Bifidobacterium species are various, belong to Bifidobacterium not of the same race form, chemical physiological characteristic, metabolism and
There are significant differences for the various aspects such as physiological function, so far, not yet studies have found that false chainlet Bifidobacterium can convert altogether
Yoke fatty acid, only minority belongs to bacterial strain not of the same race and has compared with low-conversion.Currently, also there has been no determine to lead to this for educational circles
The reason of kind function difference or mechanism.
Invention also provides application of the vacation chainlet Bifidobacterium CCFM748 in preparation conjugated linoleic acid, particularly will
Linoleic acid is conjugated linoleic acid, is especially converted into c9, t11-CLA and t10, c12-CLA, and have conversion outstanding
Rate.
The present invention also provides the vacation chainlet Bifidobacterium CCFM748 to prepare the application in conjugate linolenic acid, particularly
Conjugate linolenic acid is converted by linolenic acid, is especially converted into c9, t11, c15-CLNA and t9, t11, c15-CLNA, and have
Conversion ratio outstanding.
Application the present invention also provides the vacation chainlet Bifidobacterium PA as food additives.
False chainlet Bifidobacterium (Bifidobacterium pseudocatenulatum) CCFM748, the bacterial strain is in 2016
On June 12, in is in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, address: Chaoyang District, Beijing City north
The institute 3 of occasion West Road 1, Institute of Microorganism, Academia Sinica, deposit number are CGMCC No.12603.
[Detailed description of the invention]
Fig. 1 is separation, purifying and the preservation operational flowchart of false chainlet Bifidobacterium in the present invention；
Fig. 2 is the conversion ratio of the false chainlet Bifidobacterium CCFM748 conjugated linoleic acid of the present invention；
(A) conjugated linoleic acid total concentration with incubation time variation；
(B) distribution of culture solution, conjugated linoleic acid intracellular after cultivating 72 hours
Fig. 3 is the conversion ratio of the false chainlet Bifidobacterium CCFM748 conjugate linolenic acid of the present invention；
(A) conjugate linolenic acid total concentration with incubation time variation；
(B) culture solution, conjugate linolenic acid distribution intracellular after cultivating 72 hours
In figure: SA: stearic acid, VA: vaccenic acid, OA: oleic acid, LA: linoleic acid, CLA1:c9, t11-CLA, CLA2:t9,
t11-CLA
[specific embodiment]
It will be better understood that the present invention by following embodiments, but do not explain technical side of the invention restrictively
Case.
In the present invention, unless otherwise specified, for illustrating that " % " or percentage of concentration or ratio are weight percent
Than.
The present invention relates to following culture mediums:
MMRS fluid nutrient medium: tryptone 10g, beef extract 10g, yeast powder 5g, glucose 20g, hydrogen citrate two
Ammonium 2g, sodium acetate 5g, dipotassium hydrogen phosphate 2g, epsom salt 0.5g, Manganous sulfate monohydrate 0.25g, Tween 80 1mL and 0.5g
Cysteine adds water to 1000mL.
MMRS solid medium is to be obtained in the above basis addition with total 1.5% agar of restatement of fluid nutrient medium.
Embodiment 1: the acquisition of sample and the separation of Bifidobacterium are identified
1g Neonatal Faeces sample is taken, mMRS solid medium is coated on after gradient dilution, is placed under anaerobic environment 37
72h is cultivated at DEG C, observes and records colonial morphology, bacterium colony scribing line purifying is chosen, then in MMRS fluid nutrient medium at 37 DEG C
48h is cultivated, gained bacterium colony carries out Gram's staining and records strain morphology, and the gram negative strain in reject bacterium colony and leather are blue
Family name's positive cocci is selected to obtain Gram-positive bacillus, the reject catalase-positive organism strain after catalase is analyzed,
Retain catalase negative strain, with fructose-6-phosphate kinase assay with reject negative strain, gained passes through 16S rDNA
Sequencing is accredited as false chainlet Bifidobacterium, is named as false chainlet Bifidobacterium CCFM748.Gained vacation chainlet Bifidobacterium is passed
It is commissioned to train feeding, collects thallus and be placed in 3000r/min centrifugation 10min washing in centrifuge tube, be repeated 3 times, gained thallus is added matrix and protects
It is frozen in shield agent.
16S rDNA amplification condition: 95 DEG C of 5min；35 circulations (95 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 2min)；72℃
10min
Amplimer:
27F:(5 '-AGAGTTTGATCCTGGCTCAG-3 ')
1492R:(5 '-TACGGCTACCTTGTTACGACTT-3 '
Amplified production purifying and sequence alignment process press document (Turroni F et al.Exploring the
Diversity of the Bifidobacterial Population in the Human Intestinal Tract[J]
.Appl Environ Microb.2009；75 (6): 1534-45.) method recorded carries out.
The identification of Bifidobacterium: the Bifidobacterium isolated and purified, the method analyzed through 16S rDNA sequence are accredited as false small
Chain Bifidobacterium, fundamental characteristics are shown in Table 1.
The fundamental characteristics of table 1Bifidobacterium pseudocatenulatum CCFM748
Embodiment 2: false chainlet Bifidobacterium CCFM748 Bioconversion of conjugated linoleic acid
Specific experiment is as follows:
1, bacterial strain activates
The glycerol tube for possessing false chainlet Bifidobacterium CCFM748 is taken out from -80 DEG C of refrigerators, and bacterium solution is taken to line mMRS solid
On culture medium, the lower 37 DEG C of cultures 48h of anaerobic environment.Single colonie that picking is grown simultaneously is inoculated in mMRS fluid nutrient medium, anaerobism
The lower 37 DEG C of cultures 48h of environment, it is continuous to activate for 3 generations.
2, the preparation of linoleic acid mother liquor
It weighs 300mg linoleic acid (LA) and 200mg Tween-80 is dissolved in water and is settled to 10mL, after emulsification is sufficiently stirred,
- 20 DEG C are stored in after 0.45 μm of sterilised membrane filter filtration sterilization to be kept in dark place.
3, it is co-cultured with linoleic acid
Bacterium solution activated in step 1 is seeded to the 10mL of the LA containing 0.528mg/mL according to 2% (v/v) inoculum concentration
In mMRS fluid nutrient medium, the lower 37 DEG C of cultures of anaerobic environment 0,12,24,48,72h, while to add equivalent linoleic acid without adding
Add the culture medium of bacterium solution for control.After culture, bacterium solution is moved in centrifuge tube respectively, 5000rpm is centrifuged 5min, every part of culture
Take 3 portions of 3mL fermentation liquids stand-by to clean centrifuge tube.
4, fatty acid extracts
The extraction of fatty acid in fermentation liquid: it is extremely final concentration of that Heptadecanoic acide (C17:0) is added into 3mL fermentation liquid
Then 0.075mg/mL adds 2mL isopropanol as internal standard, sufficiently oscillation 30s；3mL n-hexane is added again, sufficiently oscillation 30s；
5000rpm is centrifuged 3min.N-hexane layer is absorbed to completely mentioning in rouge bottle, is dried with nitrogen.
The extraction of fatty acid in thallus: thallus obtained by aforementioned centrifugation with 2mL salting liquid (NaCl containing 0.137mol/L,
7.0mmol/L K2HPO4With 2.5mmoL/L KH2PO4) washing, 4000rpm centrifugation 5min, repeated washing step.By thallus weight
It is suspended from the aforementioned salting liquid of 2mL, addition Heptadecanoic acide (C17:0) to final concentration of 0.0575mg/mL is identical by fermentation liquid
Method carries out fatty acid extraction and is dried with nitrogen, and extraction obtains the fatty acid in thallus.
5, methyl esterification of fatty acid
To aforementioned fermentation liquid fatty acid, thallus fatty acid respectively with 400 μ L methanol of addition in rear are dried with nitrogen, sufficiently vibrate
Esterification is directly carried out with 150 μ L diazomethane reagents after mixing, 1mL n-hexane back dissolving is used after being dried with nitrogen, is transferred to gas phase
Bottle, pending GC-MS detection.
6, GC-MS is detected
Shimadzu gas chromatograph (GC 2010plus), gas phase column Rtx-wax (30m × 0.25mm × 0.25 μm), mass spectrograph
(Shimadzu Ultra QP2010).Temperature programming condition: initial 150 DEG C, 200 DEG C is warming up to the rate of 5 DEG C/min, is kept
10min, after with 4 DEG C/min be warming up to 230 DEG C, keep 18min.Using split sampling, 1 μ L of sample volume, split ratio 50: 1, helium
For carrier gas.Sample injector temperature and detector temperature are 240 DEG C.220 DEG C of ion source, intensity 70eV.
7, experimental result
Gained fatty acid contains: stearic acid 0.0144mg/mL, oleic acid 0.0682mg/mL, linoleic acid 0.1649mg/mL,
CLA0.3118mg/mL。
In the cumulative process of CLA, false chainlet Bifidobacterium CCFM748 is grown in the mMRS containing 0.528mg/mL LA
Start to generate CLA when 12h, with thalli growth, the content of conjugated linoleic acid is gradually increased (for 24 hours, 36h), cultivates 36h post-fermentation
The concentration of conjugated linoleic acid tends to be saturated in liquid, as shown in Figure 2.The bacterium is culture 72h or so, CLA to the maximum conversion rate of LA
Total concentration reached 0.3118mg/mL, with substrate LA total amount be calculated CLA total conversion be 59.05%.
By Analysis of Fatty Acids Composition, from the point of view of CLA Isomers content, contained only in product CLA1 (c9, t11-CLA) and
Two kinds of isomers of CLA2 (t10, c12-CLA).Bacterial strain starts to convert CLA1 after cultivating 12h.With thalli growth, isomers
CLA1 accelerated accumulation in 12h to 36h, content tends to be saturated after strain culturing 36h, and CLA2 starts to accumulate in bacterial strain
Content is lower (for 24 hours) when CLA, and with the extension of incubation time, the concentration of CLA2 is also further increased.After cultivating 72h, CLA1's
At concentrations up to for 0.223mg/mL, the 72.02% of the total CLA of Zhan.
False chainlet Bifidobacterium CCFM748 fermentation liquid and thallus fatty acid composition after cultivating 72h are analyzed it is found that the bacterium is to LA
Absorption it is high with conversion ratio, be above the prior art.It is only remained in fermentation liquid almost not residual in a small amount of substrate LA and thallus
Stay the LA being not yet converted.The gained CLA overwhelming majority is in fermentation liquid, and amount retained is also very few in thallus.Should the result shows that
Most of product is transported to extracellular not in intracellular accumulation, and CLA accounts for the ratios of total fatty acids in fermentation liquid and reaches
To 55.7%, purity is significantly higher than the prior art, because can simplify isolated purifying of the later period to CLA in fermentation liquid.
Embodiment 3: false chainlet Bifidobacterium CCFM748 bioconversion conjugate linolenic acid
1, bacterial strain activates
With embodiment 2.
2, the preparation of flax acid mother liquor
It weighs 300mg alpha-linolenic acid (α-LNA) and 200mg Tween-80 is dissolved in water and is settled to 10mL, cream is sufficiently stirred
After change, -20 DEG C are stored in after 0.45 μm of sterilised membrane filter filtration sterilization and is kept in dark place.
3, it is co-cultured with linolenic acid
Activated bacterium solution is seeded to the 10mL mMRS liquid of-LNA of α containing 0.3759mg/mL according to 2% (v/v) inoculum concentration
In body culture medium, the lower 37 DEG C of cultures of anaerobic environment 0,12,24,36,48,72h, and to add equivalent linolenic acid without adding bacterium solution
Culture medium be control.After culture, bacterium solution is moved in centrifuge tube, 5000rpm is centrifuged 5min；3 parts of 3mL are taken to every part of culture
Fermentation liquid is stand-by to clean centrifuge tube.
4, fatty acid extracts
The extraction of fatty acid in fermentation liquid: it is extremely final concentration of that Heptadecanoic acide (C17:0) is added into 3mL fermentation liquid
0.0767mg/mL makees internal standard, then adds 2mL isopropanol, sufficiently oscillation 30s；3mL n-hexane is added again, sufficiently oscillation 30s；
5000rpm is centrifuged 3min, absorbs n-hexane layer to clean and mentions rouge bottle；It is dried with nitrogen, obtains the fatty acid in fermentation liquid.
The extraction of fatty acid in thallus: thallus obtained by aforementioned centrifugation with 2mL salting liquid (NaCl containing 0.137mol/L,
7.0mmol/L K2HPO4With 2.5mmoL/L KH2PO4) wash, 4000rpm is centrifuged 5min, repeats to wash.By thallus weight
Be suspended from 2mL salting liquid, addition Heptadecanoic acide (C17:0) to final concentration of 0.0575mg/mL, by the identical method of fermentation liquid into
Row fatty acid is extracted and is dried with nitrogen, and is obtained extraction and is obtained the fatty acid in thallus.
5, methyl esterification of fatty acid
It respectively forwardly states to obtain after fatty acid is dried with nitrogen and 400 μ L methanol is added, sufficiently with 150 μ L diazonium after oscillation mixing
Methane reagent directly carries out esterification, and 1mL n-hexane back dissolving is used after being dried with nitrogen, and is transferred to gas phase bottle, pending GC-MS inspection
It surveys.
6, GC-MS is detected
Shimadzu gas chromatograph (GC 2010plus), gas phase column Rtx-wax (30m × 0.25mm × 0.25 μm), mass spectrograph
(Shimadzu Ultra QP2010).Temperature programming condition: initial 150 DEG C, 200 DEG C is warming up to the rate of 5 DEG C/min, is kept
10min, then 230 DEG C are warming up to 4 DEG C/min, keep 18min.Using split sampling, 1 μ L of sample volume, split ratio 50: 1, helium
For carrier gas.Sample injector temperature and detector temperature are 240 DEG C.220 DEG C of ion source, intensity 70eV.
7, experimental result
False chainlet Bifidobacterium CCFM748 starts to generate when growing 12h in the mMRS containing 0.3759mg/mL α-LNA
CLNA cultivates the content of conjugate linolenic acid after 36h as the content of thalli growth conjugate linolenic acid gradually increases (for 24 hours, 36h)
Increase is gradually delayed.The bacterium is culture 72h or so to the maximum conversion rate of LNA, and the total content of CLNA has reached 0.3402mg/mL (this
In outer product also contain minute quantity vaccenic acid, oleic acid, linoleic acid and substrate α-LNA), in terms of substrate α-LNA total amount its conversion
Rate is 90.5%, is significantly better than the prior art.
According to fatty acid analysis, from the point of view of CLNA Isomers content, in the tunning of the bacterium only have CLNA1 and
Two kinds of isomers of CLNA2, for the generally acknowledged highest two kinds of isomers of bioactivity.Bacterial strain starts conversion conjugation after cultivating 12h
Linolenic acid, with thalli growth, isomer C LNA1 accelerated accumulation in 12h to 36h, content tends to after strain culturing 36h
Saturation, and CLNA2 bacterial strain start accumulate CLNA when (for 24 hours) content it is lower, with the extension of incubation time, the isomers it is dense
Degree also further increases.In the CLNA isomers converted, CLNA1 accounts for absolute leading position, and ultimate density reaches 0.3267mg/
The 96% of the total CLNA content of mL, Zhan.
From Bifidobacterium CCFM748 fermentation liquid after culture 72h and thallus fatty acid composition it can be found that the bacterial strain is to LNA
Absorption it is high with conversion ratio, be significantly better than the prior art, it is remaining almost without substrate LNA in fermentation liquid, and in thallus
Also minimal amount of LNA is only remained not yet to be converted.In addition, the overwhelming majority is in fermentation liquid for the CLNA converted, bacterium
The amount retained in vivo is also considerably less.In addition, CLNA1 and CLNA2 is almost the same in fermentation liquid and endobacillary distribution situation, the knot
Fruit also indicates that most of product not in intracellular accumulation, but is transported to extracellular, therefore is conducive to the further of later period
Isolation and purification.
Claims (5)
1. application of the vacation chainlet Bifidobacterium CCFM748 in preparation conjugated linoleic acid, the vacation chainlet Bifidobacterium CCFM748
China Committee for Culture Collection of Microorganisms's General Microbiological Culture collection, preservation are preserved on June 12nd, 2016
Number CGMCC No.12603；The conjugated linoleic acid is c9, t11-CLA and t10, c12-CLA.
2. application according to claim 1, it is characterised in that the vacation chainlet Bifidobacterium CCFM748 is by linoleic acid
For conjugated linoleic acid.
3. vacation chainlet Bifidobacterium CCFM748 is preparing the application in conjugate linolenic acid, the vacation chainlet Bifidobacterium CCFM748
China Committee for Culture Collection of Microorganisms's General Microbiological Culture collection, preservation are preserved on June 12nd, 2016
Number CGMCC No.12603.
4. application according to claim 3, it is characterised in that the vacation chainlet Bifidobacterium CCFM748 converts linolenic acid
For conjugate linolenic acid.
5. application according to claim 4, it is characterised in that the conjugate linolenic acid is c9, t11, c15-CLNA and t9,
t11,c15-CLNA。
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| KR20190116248A (en) | 2017-02-10 | 2019-10-14 | 퍼펙트 (차이나) 씨오., 엘티디 | New Probiotic Bifidobacterium Strains |
| CN110106103B (en) * | 2019-03-19 | 2022-05-03 | 江南大学 | Bifidobacterium pseudocatenulatum CCFM1047, composition thereof, fermented food, application, microbial inoculum and preparation method of microbial inoculum |
| CN110093287B (en) * | 2019-03-19 | 2022-05-20 | 江南大学 | Bifidobacterium pseudocatenulatum CCFM1045, composition thereof, fermented food, application, microbial inoculum and preparation method of microbial inoculum |
| CN110106104B (en) * | 2019-03-19 | 2021-12-17 | 江南大学 | Bifidobacterium pseudocatenulatum CCFM1048, composition thereof, fermented food, application, microbial inoculum and preparation method of microbial inoculum |
| CN112695024B (en) * | 2019-10-23 | 2022-08-23 | 江南大学 | Linoleic acid isomerase and application thereof in conjugated linoleic acid production |
| CN113061169B (en) * | 2020-03-24 | 2022-09-27 | 江南大学 | Transcription regulation protein and application thereof in conjugated linoleic acid production |
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| WO2003087385A1 (en) * | 2002-04-12 | 2003-10-23 | Kabushiki Kaisha Yakult Honsha | Process for producing conjugated fatty acid and food/drink obtained by the process |
| CN1928101A (en) * | 2006-09-07 | 2007-03-14 | 河北工业大学 | Method for preparing conjugated linoleic acid |
| CN102559532A (en) * | 2010-12-13 | 2012-07-11 | 北京农业生物技术研究中心 | Strain for producing conjugated linoleic acid and application thereof |
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