CN102827899B - Gracilaria lemaneiformis agar oligosaccharide and preparation method and application thereof - Google Patents
Gracilaria lemaneiformis agar oligosaccharide and preparation method and application thereof Download PDFInfo
- Publication number
- CN102827899B CN102827899B CN2012103471055A CN201210347105A CN102827899B CN 102827899 B CN102827899 B CN 102827899B CN 2012103471055 A CN2012103471055 A CN 2012103471055A CN 201210347105 A CN201210347105 A CN 201210347105A CN 102827899 B CN102827899 B CN 102827899B
- Authority
- CN
- China
- Prior art keywords
- preparation
- agaropectin oligose
- thallus gracilariae
- agar
- membrane
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a gracilaria lemaneiformis agar oligosaccharide and the preparation method and application thereof and relates to an oligosaccharide. The invention provides a gracilaria lemaneiformis agar oligosaccharide, the preparation method of gracilaria lemaneiformis agar oligosaccharide and the application of the gracilaria lemaneiformis agar oligosaccharide in preparation of antioxidant and uvioresistant health care products and cosmetics. Pacific heating color bacillus H2 and gracilaria lemaneiformis are co-cultivated and the gracilaria lemaneiformis is directly degraded by the enzyme system which is generated by bacterial strain to obtain the gracilaria lemaneiformis agar oligosaccharide of which the degree of polymerization is 4-8. The gracilaria lemaneiformis agar oligosaccharide has the physiological activities of antioxidation, absorbing ultraviolet rays and the like. In preparation, the bacterial strain is activated by an activation medium plate and inoculated in a seed medium; fermentation broth is obtained by shake cultivation and inoculated in a saccharide generating medium; after shaking fermental cultivation and centrifugalization, supernate is collected; the supernate is successively filtered by a membrane filtration devices with a 0.45mu m filter membrane, a 0.22mu m filter membrane, a 10000-50000D ultrafiltration membrane and a 300-600D ultrafiltration membrane, and the percolate is the gracilaria lemaneiformis agar oligosaccharide.
Description
Technical field
The present invention relates to a kind of oligosaccharides, relate to a kind of Thallus Gracilariae agaropectin oligose and preparation method thereof, this oligosaccharides utilizes the common cultivation of bacterial strain and Thallus Gracilariae to be prepared from, and has the physiologically actives such as anti-oxidant, antiultraviolet.
Background technology
Oligosaccharides refers to the low polymerization glucide of the straight or branched consisted of 2～20 identical or different monose, claims again oligose, comprises common oligosaccharides and the large class of functional oligosaccharide two.Sucrose that people are familiar with, maltose, lactose etc. are arranged in common oligosaccharides, these sugar can be digested and assimilated by body, intestinal beneficial bacterium be there is no to growth-promoting effect, another kind of is functional oligosaccharide, such as Nutriflora P, xylo-oligosaccharide, soybean oligosaccharide, oligochitosan, agaropectin oligose, algin oligosaccharide etc., they itself can not be digested and assimilated by body, but have multiple physiologically active, in fields such as food, agriculturals, the wide market space are arranged.
Alga oligosaccharide is found to have important physiologically active in recent years, wherein from the agaropectin oligose of the red algaes such as river hedge, gelidium, studies comparatively extensively.The agaropectin oligose of for example advocating in Chinese patent CN200410024380.9 has the effect of prebiotics; the agar oligosaccharide mixture of advocating in Chinese patent CN03138973.2 patent can be used as the hypoglycemic medicine of preparation or foodstuff additive, and the agar-agar oligosaccharide of advocating in Chinese patent CN1171592C patent has fibroblastic growth promoter action and ultraviolet oxidative damage provide protection etc.
But the application of agaropectin oligose is still less so far, the bottleneck factor that affects its application and development mainly is the preparation method.The preparation process of agaropectin oligose is all to extract agar-agar in the red algaes such as first Congjiang hedge, gelidium at present, and then with chemical method or enzyme process, agar-agar is degraded to the oligosaccharides that molecular weight is less.Chinese patent CN01115094.7 for example, CN021132573.1, CN03138971.6 philosophy advocate to adopt different chemical process degraded agar-agars to obtain oligose; Chinese patent CN03112515.8, CN03112518.2, the opinions such as CN200310114439.9 utilize the agarase preparation that natural bacterium or genetic engineering bacterium obtain to carry out preparation of agar oligosaccharide etc.But be that chemical method or enzymolysis process all exist comparatively serious shortcoming, comprise: 1. these methods all can only act on the Sargassum polysaccharides such as agar-agar, and need pass through alkali, acid processing and washing repeatedly repeatedly from marine alga, extracting Sargassum polysaccharides, caused serious pollution and the consumption of water resources; 2. enzymolysis process need, first through the extensive preparation of zymin, then just can be applied to the degraded of polysaccharide.But the preparation of zymin requires highly to plant and instrument, has improved production cost, and has made integrated artistic complicated; 3. still lack at present efficient agarase, cause agaropectin oligose can't obtain scale operation and application; 4. the chemical method agaropectin oligose molecular weight heterogeneity of producing, unstable product quality, be difficult to carry out scale operation.The current agarase that only has the U.S. to use Atlantic Ocean pseudomonas prepares the product of agar oligosaccharide, but expensive, can't meet the requirement of development and production.
China's algal cultivation is in large scale, and Thallus Gracilariae involved in the present invention is a kind of Gracilaria marine alga, is also one of main algal cultivation kind of China, and its economic worth mainly comes to be extracted agar-agar and, as the abalone feed, be in the devalued stage of utilizing.The method that the present invention adopts bacterial strain and Thallus Gracilariae to cultivate altogether, utilize the enzyme system direct degraded Thallus Gracilariae that Institute of Micro-biology produces to obtain agaropectin oligose, can solve the problems referred to above in the application and development of restriction agaropectin oligose, realizes the higher value application of Thallus Gracilariae.
Summary of the invention
The object of the present invention is to provide a kind of Thallus Gracilariae agaropectin oligose and preparation method thereof.
The 2nd purpose of the present invention is to provide that the Thallus Gracilariae agaropectin oligose is anti-oxidant in preparation, the application in uvioresistant healthcare products and makeup.
Described Thallus Gracilariae agaropectin oligose is to utilize Pacific Ocean heat color bacillus (Flammeovirga Pacifica) H2 and Thallus Gracilariae to cultivate altogether, utilizing the enzyme that bacterial strain produces is that the direct degraded Thallus Gracilariae acquisition polymerization degree is 4～8 agaropectin oligose, has the physiologically actives such as anti-oxidant, that absorption is ultraviolet.
Described Pacific Ocean heat color bacillus (Flammeovirga Pacifica) H2 is separated and obtains voluntarily by the inventor, and on June 13rd, 2012, be preserved in Chinese Typical Representative culture collection center, deposit number is CCTCC NO:M2012229, and address is Wuhan, China university (postcode 430072).The source of sample be nero deep 5378m depths the mud sample (E157 ° 24 ' 31 ", N19 ° of 30 ' 30 ").The screening and separating of bacterial strain and cultural method are: abyssal sediment 2g is put into to the 2216E liquid nutrient medium that 30mL contains 1% gardon asparagus powder of sieving through 60～80 orders, enrichment in 4 ℃.Then by 1000 times of pregnant solution dilutions, coat and contain 1% on the 2216E flat board of the gardon asparagus powder that 60～80 orders sieve, in 16 ℃ of cultivations.The bacterial strain obtained is further purified to conservation, and identifies.
The preparation method of Thallus Gracilariae agaropectin oligose of the present invention is as follows:
1) after bacterial strain Pacific Ocean heat color bacillus (Flammeovirga Pacifica) H2 is activated with the activation medium flat board, be inoculated in seed culture medium, shaking culture obtains fermented liquid;
2) fermented liquid obtained in step 1) is inoculated in the product sugar culture-medium, centrifugal after the continuation oscillation and fermentation cultivation, collect supernatant liquor;
3) adopt membrane filter plant, supernatant liquor is used to 0.45 μ m successively, 0.22 μ m filter membrane, 10000～50000D ultra-filtration membrane and 300～600D nanofiltration membrane, the gained filtrate is the Thallus Gracilariae agaropectin oligose.
In step 1), described activation medium flat board can be: contain the gardon asparagus powder 1% after 60～80 orders sieve, and yeast extract paste 0.2%, NaCl3%, agar-agar powder 1.5%, the distilled water preparation, through 121 ℃, the sterilizing rear use of 20min; Described seed culture medium can be: contain CMC-Na
20.5%～1.5%, yeast extract paste 0.2%, NaCl3%, the distilled water preparation, through 121 ℃, the sterilizing rear use of 20min; The condition of described shaking culture can be: under 25～37 ℃ of conditions, the 250r/min shaking culture is to OD
600=1.0～1.5.
In step 2) in, described product sugar culture-medium can be: contain the gardon asparagus powder 1%～5% of sieving through 60～80 orders, MgCl
20.1%～0.5%, the distilled water preparation, through 121 ℃, the sterilizing rear use of 20min; The inoculative proportion of fermented liquid can be 5%; The condition of described oscillation and fermentation cultivation can be: under 25～37 ℃ of conditions, and 300r/min oscillation and fermentation cultivation 24～72h.
In step 3), the centrifugal condition of described fermented liquid can be: the centrifugal 15min of 10000 * g; Described filter membrane can be the rolling organic membrane.
Thallus Gracilariae agaropectin oligose of the present invention can be applied in anti-oxidant, the uvioresistant healthcare products of preparation and makeup.
The agaropectin oligose of Thallus Gracilariae described in the present invention preparation technology's advantage is:
1. adopt bacterial strain and Thallus Gracilariae to cultivate altogether, the enzyme system that utilizes bacterial strain to produce, single stage method directly obtains agaropectin oligose.And enzymolysis process described in existing research report or patent prepares oligosaccharides and all needs a plurality of steps, mainly comprise by producing the agarase strain fermentation and prepare agarase, extract agar-agar, agarase reacted to obtain oligosaccharides etc. with agar-agar by marine algas such as Thallus Gracilariaes.Technique in the present invention has obvious novelty, and technique is simple, pollution-free, cost is low, efficiency is high, has good practicality.
2. in producing sugar culture-medium, utilize the MgCl of lower concentration
2Replace NaCl(concentration commonly used in fermentation generally more than 1%), not only reduce the corrosion to fermentation equipment, and reduced the cost of alga oligosaccharide subsequent purification.
The physiologically active of the agaropectin oligose of Thallus Gracilariae described in the present invention is characterized as:
Have significant oxidation-resistance, comprise CCl
4The acute liver caused, and the fruit bat death that Paraquat causes has significant protective effect; Have that very strong absorption is short, the activity of ultraviolet B radiation.In current disclosed alga oligosaccharide Patents, the alga oligosaccharide that CN1843325A advocates is algin oligosaccharide, is mainly used in that makeup are sun-proof, the activity of moisturizing; The alga oligosaccharide that CN1843153A advocates is mainly used in fodder additives; The agaropectin oligose that CN1593433A advocates as prebiotics for animal and human's body.The activity of the agaropectin oligose therefore, obtained in the present invention is showed no other research report or patent.Although the alga oligosaccharide that CN1843325A advocates is mentioned, have sunscreen actives, active relevant to uv-absorbing, the described alga oligosaccharide of this patent is algin oligosaccharide, not agaropectin oligose described in the present invention.In sum, in the present invention, prepared agaropectin oligose has obvious creativeness and practicality.
The accompanying drawing explanation
Fig. 1 is the thin layer chromatography analysis of asparagus active oligosaccharides.In Fig. 1, M: be followed successively by from top to bottom and be respectively Neoagarotetraose, neoagarohexaose and neoagarooctaose standard substance; 1,2: the Thallus Gracilariae agaropectin oligose.
Fig. 2 is liver section HE coloration result.
Fig. 3 is Thallus Gracilariae agaropectin oligose full wavelength scanner figure.In Fig. 3, X-coordinate is wavelength (nm), and ordinate zou is absolute strength Abs.
Embodiment
Below by embodiment, the present invention is described in further detail, but the present invention not only is confined to following examples.
The microbe fermentation method preparation of embodiment 1 Thallus Gracilariae agaropectin oligose
1) bacterial strain Pacific Ocean heat color bacillus (Flammeovirga Pacifica) H2 (is contained to the gardon asparagus powder 1% of sieving through 60～80 orders with activation medium, yeast extract paste 0.2%, NaCl3%, agar-agar powder 1.5%, the distilled water preparation, pH7～8, used after 121 ℃ of sterilizing 20min) activate after, picking list colony inoculation (contains CMC-Na in seed culture medium
21%, yeast extract paste 0.2%, NaCl3%, the distilled water preparation, pH7～8, used after 121 ℃ of sterilizing 20min) in, shaking culture is to OD under the condition of 30 ℃
600=1.0;
2) fermented liquid is added and produces sugar culture-medium and (contain the gardon asparagus powder 3% of sieving through 60～80 orders, MgCl according to 3% inoculative proportion
20.25%, the distilled water preparation, pH7～8, used after 121 ℃ of sterilizing 20min) in, continue oscillation and fermentation cultivation 48h under 30 ℃ of conditions;
3) by fermented liquid through the centrifugal 15min of 10000 * g, collect supernatant liquor;
4) adopt membrane filter plant, supernatant liquor is used to 0.45 μ m successively, 0.22 μ m filter membrane, 30000D ultra-filtration membrane and 300D nanofiltration membrane, the gained filtrate is the Thallus Gracilariae agaropectin oligose;
5) by the DNS method, measure oligosaccharide concentration, preserve in 10 ℃ of following temperature.
The thin-layer chromatogram of gained Thallus Gracilariae agaropectin oligose composition is referring to Fig. 1, and prepared oligosaccharides is the mixture of 4～8 sugar.
Below provide the experimental verification of the anti-oxidant physiologically active of gained Thallus Gracilariae agaropectin oligose.
1. the provide protection of Thallus Gracilariae agaropectin oligose to the mouse carbon tetrachloride acute liver damage
Mouse peritoneal injection CCl
4After, CCl
4In liver, produce free radical under the catalysis of enzyme and make cytolemma generation lipid peroxidation, thereby destroy the integrity of membrane structure and function, enzyme in tenuigenin is infiltrated in blood, cause activity of serum enzyme to change, cell generation balloon sample is become and necrosis, this process is optionally destroyed the liver cell of leaflet central area.
Experimentation on animals:
Laboratory animal and grouping: select healthy male, BALB/c mouse, 6 ~ 7 weeks are big or small, body weight 18 ~ 20g.Method by random packet is divided into 4 groups by mouse, 3 every group.The 1st group is high dosage Thallus Gracilariae agaropectin oligose administration group, and the 2nd group is low dosage Thallus Gracilariae agaropectin oligose administration group, and the 3rd group is the blank group, and the 4th group is CCl
4The damage control group.
Dosage and method: A group perfusion amount is 200mg/ (kg.BW), and B group perfusion amount is 150mg/ (kg.BW), and C, D organize and fill with the equivalent sterile purified water every day.Each organized continuous irrigation 7 days, after 7 days, A, B, D group mouse was pressed to the consumption abdominal injection CCl of 1mL/kg body weight
4(according to the ratios of 2: 5 by CCl
4Be diluted in whiteruss), after injection 48h, eyeball is got blood system from serum, dissects and gets liver analysis.
The mensuration of mouse observation index: MAIN OUTCOME MEASURES is Mouse Weight, liver weight, Serum ALT content and hepatic pathology tissue examination.Mouse Weight is weighing before getting blood, and liver carries out weighing after focusing on dissection.ALT content in serum adopts reitman-frankel method to measure, and 50 times of left and right of serum dilution are recorded to its absorbance with the ALT test kit afterwards; The reitman-frankel method unit conversion obtained is returned to unit of enzyme IU/L, 1 Ka Menshi unit under 25 ℃ of conditions=0.482I U/L.And the hepatic pathology tissue examination adopts hematoxylin-eosin (HE) staining to observe, choose after the mouse liver tissue is made paraffin section and carry out under Electronic Speculum, observing after HE dyeing.
Experimental result:
By measuring body weight, the liver of respectively organizing mouse, weigh and the serum alt result, obtain the data as table 1:
Table 1
By the weight ratio to each group mouse, can see, wherein A group 200mg/ (kg.BW) Mouse Weight is on average higher than all the other Mouse Weights of several groups, and the weight average of B group 150mg/ (kg.BW) mouse is lower than all the other groups, the alga oligosaccharide that 200mg/ (kg.BW) concentration is described has promoter action to the body weight of mouse, and the alga oligosaccharide of 150mg/ (kg.BW) concentration has opposite effect to the body weight of mouse.And can obtain similar Mouse Weight result relatively by the liver anharmonic ratio to each group mouse.To ALT result in each group mice serum relatively find taken the ALT level of experimental mice of alga oligosaccharide generally lower than CCl
4The damage control group, wherein the ALT content of A group 200mg/ (kg.BW) mouse is basic consistent with the level of blank group, and B organizes the level of 150mg/ (kg.BW) mouse ALT content a little more than the blank group, but also well below CCl
4The damage control group.Illustrate that alga oligosaccharide is for CCl
4The liver injury caused has good repairing effect.
And the liver organization of each group mouse is carried out to paraffin embedding, and then carry out HE dyeing, after observed under electron microscope, take a picture and preserve referring to Fig. 2.According to liver section HE coloration result, can find CCl
4The circumvascular cell injury of the hepatic tissue that can cause, and the circumvascular cell of hepatic tissue the not damaged of A group 200mg/ (kg.BW) mouse, and the circumvascular cell of B group 150mg/ (kg.BW) murine liver tissue has damage, body weight and the heavy lighter reason of liver that this has also explained B group 150mg/ (kg.BW) mouse, illustrate the CCl of the alga oligosaccharide of 150mg/ (kg.BW) concentration to mouse
4The hepar damnification repair caused does not have the good of 200mg/ (kg.BW) concentration, but the alga oligosaccharide of 200mg/ (kg.BW) concentration is to the CCl of mouse
4The hepar damnification repair successful caused.Illustrate that alga oligosaccharide is for CCl
4The liver injury caused has good repairing effect.
2. the fruit bat of take detects the oxidation-resistance of Thallus Gracilariae agaropectin oligose to the resistance of Paraquat as model
(1) detection method
With the pyrogallol Autoxidation Method, measure.With 10mM HCl preparation 0.1mM pyrogallol, by table 2, get 50mMTris-HCl damping fluid (pH8.0), 10mM HCl, the Thallus Gracilariae agaropectin oligose, after mixing, at 25 ℃ of water bath heat preservation 20min, after taking-up, add immediately the 0.1mM pyrogallol (cumulative volume 3mL) of 25 ℃ of preheatings of 0.4mL, shake up rapidly, pour cuvette into, measure OD
325Value.
Table 2
| Reaction system | Tris-HCl（mL） | HCl（mL） | Thallus Gracilariae agaropectin oligose (mL) |
| Blank | 2 | 1 | 0 |
| Contrast | 2 | 0.6 | 0 |
| A1* | 2 | 0.3 | 0.3 |
| A2* | 2 | 0.2 | 0.4 |
| A3* | 2 | 0.1 | 0.5 |
* A1 oligosaccharides final concentration is 170mg/L; A2 oligosaccharides final concentration is 227mg/L; A3 oligosaccharides final concentration is 283mg/L.
(2) experimental result
Inhibiting rate calculation formula: inhibiting rate=(A
Right-A
Place)/A
Right
In formula, A
RightFor ..., A
PlaceFor ...
Table 3
| Process | Regression equation | R2 | △OD/min | Inhibiting rate (%) |
| Contrast | y=0.0418x+0.002 | 0.9973 | 0.0852 | 0 |
| A1 | y=0.0257x+1.1369 | 0.9874 | 0.0586 | 31.22 |
| A2 | y=0.0267x+1.1208 | 0.9938 | 0.0565 | 33.69 |
| A3 | y=0.0262x+1.4755 | 0.9981 | 0.0523 | 38.62 |
By table 3, illustrated: the Thallus Gracilariae agaropectin oligose can obviously slow down the autooxidation of pyrogallol, and inhibiting rate can reach 30%, and the higher restraining effect of concentration is stronger.
3. the fruit bat of take is that model detects the oxidation resistant practical application effect of Thallus Gracilariae agaropectin oligose to the resistance of Paraquat
(1) experimental technique: female, male each 420 of the wild-type OR fruit bat of getting firm emergence, its male and female are separately raised, after 4～7 days, test.6 metafiltration paper are laid in bottom in flat based tubes, in each flat-ended tube, add the processing sample of 500 μ L, and 6 experimental group (referring to table 4) are set, each 3 repetitions of each experimental group male and female, and each repeats to place 20 fruit bats.Every 12h, record the death condition of fruit bat.Record fruit bat mortality ratio after 108h.
(2) experimental result:
Paraquat can act on the redox reaction of cell, and activation is its toxic action basis for oxyradical in cell, formed excessive super oxide anion free radical (O2-) and hydrogen peroxide (H
2O
2) etc. can cause the peroxidation of many histoorgan Cell membrane lipids, thereby cause the infringement of multisystem histoorgan.The Thallus Gracilariae agaropectin oligose can obviously reduce the mortality ratio of fruit bat, and especially female fruit bat shows that the Thallus Gracilariae agaropectin oligose has the effect of removing superoxide radical.
Table 4 be take fruit bat and is detected the oxidation-resistance of Thallus Gracilariae agaropectin oligose as model
| Process | Female fruit bat mortality ratio (%) | Male drosophila mortality ratio (%) |
| Sucrose | 0 | 1.3 |
| Sucrose+Paraquat | 90 | 91.7 |
| Sucrose+oligosaccharides (0.2mg/mL) | 0 | 1.7 |
| Sucrose+oligosaccharides (0.2mg/mL)+Paraquat | 11.7 | 75 |
Below provide the detection example that gained Thallus Gracilariae agaropectin oligose absorbs ultraviolet performance
(1) measuring method: use the UV1800 type uv-spectrophotometric instrument of SHIMADZU company, the Thallus Gracilariae agaropectin oligose is added in cuvette, with the distilled water zeroing, according to following parameter measurement: measurement pattern ABS; Sweep limit 800～200nm; Recording interval-4.000～4.000A; Scanning step 0.5nm.
(2) measurement result:
Wavelength all belongs to ultraviolet ray range on spectrum in the radiation between 100～400nm, therefore be called as ultraviolet radiation, and by wavelength division, generally be divided into 3 kinds: the UVC(short wave ultraviolet), wavelength is 100～280nm; The UVB(ultraviolet B radiation), wavelength is 280～320nm; The UVA(long wave ultraviolet), wavelength is 320～400nm.
In these 3 kinds of ultraviolet rays, UVC is blocked in outside earth's surface by ozonosphere.10% even lower UVB radiation can reach earth's surface, is one of reason caused skin aging, sunburn, immunosuppression, DNA damage and skin carcinoma.UVA radiation more than 90% can reach earth's surface, and its sunburn to skin and destructive intensity are not so good as UVB, but can suppress the immunity system function of skin.
As can be seen from Figure 3, the absorption value of Thallus Gracilariae agaropectin oligose in 230～310nm scope is the highest, shows the maximum UVB of harm in ultraviolet ray is had to good sorption, can be used for the exploitation of ultra-violet radiation resisting product.
Embodiment 2
Similar to Example 1, its difference is in step 1) and 2) described in culture temperature be 25 ℃, CMC-Na in seed culture medium described in step 1)
2Content be 0.5%, step 2) in gardon asparagus powder be 1%, MgCl
2Content is 0.1%, and incubation time is 72h.
Embodiment 3
Similar to Example 1, its difference is in step 1) and 2) described in culture temperature be 37 ℃, CMC-Na in seed culture medium described in step 1)
2Content be 1.5%, step 2) in gardon asparagus powder be 5%, MgCl
2Content is 0.5%, and incubation time is 24h.
Embodiment 4
Similar to Example 1, its difference is in step 1) and 2) described in culture temperature be 30 ℃, step 2) in gardon asparagus powder be 1%, MgCl
2Content is 0.1%, and incubation time is 72h.
Embodiment 5
Similar to Example 1, its difference is in step 1) and 2) described in culture temperature be 25 ℃, step 2) in MgCl
2Content is 0.5%, and incubation time is 48h.
Embodiment 6
Similar to Example 1, its difference is in step 1) and 2) described in culture temperature be 30 ℃, CMC-Na in seed culture medium described in step 1)
2Content be 1.5%, step 2) in gardon asparagus powder be 2%, MgCl
2Content is 0.4%, and incubation time is 48h.
Embodiment 7
Similar to Example 1, its difference is in step 1) and 2) described in culture temperature be 37 ℃, CMC-Na in seed culture medium described in step 1)
2Content be 0.5%.
Claims (6)
1. the preparation method of a Thallus Gracilariae agaropectin oligose is characterized in that comprising the following steps:
1) after bacterial strain Pacific Ocean heat color bacillus (Flammeovirga Pacifica) H2 is activated with the activation medium flat board, be inoculated in seed culture medium, shaking culture obtains fermented liquid; Described activation medium flat board is: contain the gardon asparagus powder 1% after 60～80 orders sieve, and yeast extract paste 0.2%, NaCl 3%, agar-agar powder 1.5%, the distilled water preparation, through 121 ℃, the sterilizing rear use of 20min; Described seed culture medium is: contain CMC-Na
20.5%～1.5%, yeast extract paste 0.2%, NaCl 3%, the distilled water preparation, through 121 ℃, the sterilizing rear use of 20min; Described Pacific Ocean heat color bacillus (Flammeovirga Pacifica) H2 is preserved in Chinese Typical Representative culture collection center on June 13rd, 2012, and deposit number is CCTCC NO:M2012229;
2) fermented liquid obtained in step 1) is inoculated in the product sugar culture-medium, centrifugal after the continuation oscillation and fermentation cultivation, collect supernatant liquor; Described product sugar culture-medium is: contain the gardon asparagus powder 1%～5% of sieving through 60～80 orders, MgCl
20.1%～0.5%, the distilled water preparation, through 121 ℃, the sterilizing rear use of 20min;
3) adopt membrane filter plant, supernatant liquor is used to 0.45 μ m successively, 0.22 μ m filter membrane, 10000～50000D ultra-filtration membrane and 300～600D nanofiltration membrane, the gained filtrate is the Thallus Gracilariae agaropectin oligose.
2. a kind of preparation method of Thallus Gracilariae agaropectin oligose as claimed in claim 1, is characterized in that in step 1), and the condition of described shaking culture is: under 25～37 ℃ of conditions, the 250r/min shaking culture is to OD
600=1.0～1.5.
3. a kind of preparation method of Thallus Gracilariae agaropectin oligose as claimed in claim 1, is characterized in that in step 2) in, the inoculative proportion of described fermented liquid is 5%.
4. a kind of preparation method of Thallus Gracilariae agaropectin oligose as claimed in claim 1, is characterized in that in step 2) in, the condition of described oscillation and fermentation cultivation is: under 25～37 ℃ of conditions, and 300r/min oscillation and fermentation cultivation 24～72h.
5. a kind of preparation method of Thallus Gracilariae agaropectin oligose as claimed in claim 1, is characterized in that in step 3), and the centrifugal condition of described fermented liquid is: the centrifugal 15min of 10000 * g.
6. a kind of preparation method of Thallus Gracilariae agaropectin oligose as claimed in claim 1, is characterized in that in step 3), and described filter membrane is the rolling organic membrane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103471055A CN102827899B (en) | 2012-09-18 | 2012-09-18 | Gracilaria lemaneiformis agar oligosaccharide and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103471055A CN102827899B (en) | 2012-09-18 | 2012-09-18 | Gracilaria lemaneiformis agar oligosaccharide and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102827899A CN102827899A (en) | 2012-12-19 |
| CN102827899B true CN102827899B (en) | 2013-11-27 |
Family
ID=47331207
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012103471055A Active CN102827899B (en) | 2012-09-18 | 2012-09-18 | Gracilaria lemaneiformis agar oligosaccharide and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102827899B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI632359B (en) * | 2017-07-21 | 2018-08-11 | 嘉藥學校財團法人嘉南藥理大學 | Method for rapid determination of toxic substances by plant gel coagulation |
Families Citing this family (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103114056B (en) * | 2013-01-23 | 2015-04-22 | 国家海洋局第三海洋研究所 | Aldehyde dehydrogenase and preparation method thereof |
| CN103352045B (en) * | 2013-06-28 | 2015-05-13 | 国家海洋局第三海洋研究所 | Arylsulfatase and preparation method and applications thereof |
| CN104150977B (en) * | 2014-07-25 | 2017-02-15 | 国家海洋局第三海洋研究所 | Preparation method and application of alga oligosaccharide biological fertilizer |
| CN104164461A (en) * | 2014-07-25 | 2014-11-26 | 国家海洋局第三海洋研究所 | Method for raising asparagus oligosaccharide output by addition of metal ions |
| CN105533167A (en) * | 2015-12-08 | 2016-05-04 | 福建省农业科学院中心实验室 | Alga oligosaccharide fish feed additive |
| CN105505835B (en) * | 2016-01-25 | 2019-09-27 | 集美大学 | A kind of method and its application improving streptococcus thermophilus activity and alimentary canal tolerance |
| CN106173201A (en) * | 2016-07-04 | 2016-12-07 | 汕头大学 | A kind of fermentation process improving Thallus Gracilariae quality |
| CN106173516A (en) * | 2016-07-04 | 2016-12-07 | 汕头大学 | A kind of black porgy feedstuff containing fermentation Thallus Gracilariae and preparation method thereof |
| CN106420471B (en) * | 2016-10-31 | 2017-08-25 | 广东科玮生物技术股份有限公司 | It is a kind of to have sun-proof and after-sun function skin care item and preparation method thereof concurrently |
| CN107087641B (en) * | 2017-02-08 | 2021-04-13 | 福建三炬生物科技股份有限公司 | Marine oligosaccharide biological preparation for improving salt resistance of crops and preparation method thereof |
| CN109125179B (en) * | 2018-10-15 | 2021-04-30 | 华侨大学 | Antioxidant soothing moisturizing cream composition and preparation method thereof |
| CN109820038A (en) * | 2019-04-03 | 2019-05-31 | 福州大学 | A kind of agar Yoghourt and preparation method thereof |
| CN112159827B (en) * | 2020-10-16 | 2022-03-08 | 福建环海生物科技股份有限公司 | Production process of agar oligosaccharide |
| CN112704230A (en) * | 2021-01-08 | 2021-04-27 | 福建保安康保健食品有限公司 | Agar oligosaccharide powder product and application thereof |
-
2012
- 2012-09-18 CN CN2012103471055A patent/CN102827899B/en active Active
Non-Patent Citations (6)
| Title |
|---|
| 刘刚.琼胶寡糖的制备及生物活性研究.《中国优秀硕士学位论文，医药卫生科技辑》.2012,E055-22. * |
| 毛文君.琼胶寡糖的制备及其13C-NMR研究.《水产学报》.2001,第25卷(第6期),p582-584. |
| 琼胶寡糖的制备及其13C-NMR研究;毛文君;《水产学报》;20011231;第25卷(第6期);p582-584 * |
| 琼脂寡糖的制备及其应用研究进展;问莉莉;《山东化工》;20110531;第40卷(第5期);p28-30 * |
| 赵永强.龙须菜琼胶糖的制备及其副产物的研究.《中国优秀硕士学位论文,工程科技I辑》.2011,B024-10. * |
| 问莉莉.琼脂寡糖的制备及其应用研究进展.《山东化工》.2011,第40卷(第5期),p28-30. |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI632359B (en) * | 2017-07-21 | 2018-08-11 | 嘉藥學校財團法人嘉南藥理大學 | Method for rapid determination of toxic substances by plant gel coagulation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102827899A (en) | 2012-12-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102827899B (en) | Gracilaria lemaneiformis agar oligosaccharide and preparation method and application thereof | |
| El-Gendi et al. | Recent advances in bacterial cellulose: a low-cost effective production media, optimization strategies and applications | |
| Anbuchezhian et al. | Prospect of marine algae for production of industrially important chemicals | |
| CN101643759B (en) | Method for preparing Schizophyllum commune Fr polysaccharides and dedicated culture medium thereof | |
| CN104130958B (en) | One bacillus subtilis and the application in preventing and treating root knot nematode on tree peony disease thereof | |
| CN109694416A (en) | A kind of high-purity, highly dissoluble, high activity edible fungi polysaccharide preparation method | |
| CN108823261A (en) | A kind of Ultra-low molecular weight Dendrobium officinale polysaccharide and its preparation and application | |
| CN104099395A (en) | Rhodobacter sphaeroides extracting solution with antioxidant activity and preparation method thereof | |
| CN105695347A (en) | Strain producing pullulan, application thereof and pullulan production method | |
| CN104498398A (en) | Lactobacillus plantarum with function of removing patulin | |
| CN104403973A (en) | Digestive lactobacillus alimentris with function of removing patulin and application of digestive lactobacillus | |
| CN102552335A (en) | Traditional Chinese medicine health care product, its preparation method and its application | |
| CN107409742A (en) | A kind of artificial culture method of cicada fungus | |
| CN102229894B (en) | Plectosphaerella cucumerina HL-02 and use thereof in preparation of D-lactonohydrolase | |
| CN102268467A (en) | Polysaccharide extract of Lactarius deliciosus hypha and application thereof | |
| CN101717731B (en) | Phaffiarhodozyma mutant strain MK19 of glucose metabolism de-repression high-yield astaxanthin and culturing method thereof | |
| CN104799286A (en) | Composition for improving immunity and preparation method of composition | |
| CN109699394A (en) | A method of vertical stripe Xylaria sp. fungus fructification is produced using fluid nutrient medium | |
| CN112704230A (en) | Agar oligosaccharide powder product and application thereof | |
| CN103911321A (en) | Cypermethrin- and/or deltamethrin- degrading bacterium in seawater environment, and separation, purification and applications thereof | |
| EP2455448A1 (en) | Pharmaceutical composition for inhibiting infection and replication of influenza A and B virus, and the manufacture thereof | |
| CN109619591A (en) | A kind of fingered citron pectin oligosaccharide and its preparation and application | |
| CN106636252B (en) | Wizened bacterium exopolysaccharide and its preparation method and application | |
| CN109007790A (en) | A kind of preparation method of Chinese yam mycoplasma powder and its Chinese yam mycoplasma powder of preparation | |
| CN102373166B (en) | Bacillus thuringiensis PX-95 strain capable of producing poly-beta-hydroxybutyric acid at high yield and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 361005 Fujian Province University of Siming District of Xiamen City Road No. 178 Patentee after: THIRD INSTITUTE OF OCEANOGRAPHY, MINISTRY OF NATURAL RESOURCES Address before: 361005 Fujian Province University of Siming District of Xiamen City Road No. 178 Patentee before: THIRD INSTITUTE OF OCEANOGRAPHY, STATE OCEANIC ADMINISTRATION |
|
| CP01 | Change in the name or title of a patent holder | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230927 Address after: 361000 room 201B, plant 8, No. 3701, Xiang'an North Road, Xiamen Torch High tech Zone (Xiang'an) Industrial Zone, Xiamen, Fujian Patentee after: LANNAO TECHNOLOGY (XIAMEN) Co.,Ltd. Address before: 361005 Fujian Province University of Siming District of Xiamen City Road No. 178 Patentee before: THIRD INSTITUTE OF OCEANOGRAPHY, MINISTRY OF NATURAL RESOURCES |
|
| TR01 | Transfer of patent right |