CN1086736C - Microorganism carrier fixed with glue compounded with carragheen and konjak polysaccharide and use thereof - Google Patents
Microorganism carrier fixed with glue compounded with carragheen and konjak polysaccharide and use thereof Download PDFInfo
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- CN1086736C CN1086736C CN99113707A CN99113707A CN1086736C CN 1086736 C CN1086736 C CN 1086736C CN 99113707 A CN99113707 A CN 99113707A CN 99113707 A CN99113707 A CN 99113707A CN 1086736 C CN1086736 C CN 1086736C
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- konjac polysaccharide
- carrageenin
- gum blend
- polysaccharide gum
- microorganism carrier
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Abstract
The present invention discloses a microorganism carrier immobilized with carrageenan-konjak polysaccharide compounded glue, and applications thereof in the production process of the preparation of nucleoside triphosphates, such as adenosine triphosphate (ATP), guanosine triphosphate (GTP), cytidine triphosphate (CTP), uridine triphosphate (UTP), etc. The present invention has the advantages of high intensity of microorganism immobilization, high elasticity, difficult crushing, difficult decomposition by microorganisms, high substrate conversion rate, high product purity, etc. The present invention is favorable for repeated use for a long term.
Description
The invention belongs to the biochemical engineering technical field, relate to a kind of Immohilized microorganism carrier and application thereof, relate in particular to a kind of carrageenin-konjac polysaccharide gum blend Immohilized microorganism carrier, and the application of this fixed microorganism carrier in preparation ribonucleoside triphosphote such as adenosine triphosphate (ATP), black guanosine triphosphate (GTP), cytidine triphosphate(CTP) (CTP) and uridine triphosphate production processes such as (UTP).
Ribonucleoside triphosphote is a high energy compound in the organism, can be used as diseases such as biochemical drug treatment amyotrophy and cardiac disorder, cytidine triphosphate(CTP) (CTP) can also be treated cerebral concussion, cerebral trauma and serious neurasthenia etc., and in addition, ribonucleoside triphosphote still is a kind of important biochemical reagents.Preparing ribonucleoside triphosphote with immobilized microorganism is the inchoate new production technology seventies.Chinese patent ZL 92113813.X discloses a kind of method with gelatin glutaraldehyde Immobilized Yeast adenosine triphosphate (ATP), fixed compound can use repeatedly though this method has, separation and Extraction simple and the product purity advantages of higher, but because gelatin is a kind of protein, easily degraded by the proteolytic enzyme in the microorganism, so after it is subjected to living contaminants or life-time service, immobilization particle intensity and proportion can reduce, and will increase operation easier and influence work-ing life.
Japan has reported a kind of method of utilizing the dry bread yeast to make ribonucleoside triphosphote in 1978 56 volumes of fermentation engineering the 5th phase manger storehouse, but the transformation efficiency of its adenosine triphosphate (ATP) only is 53%, the transformation efficiency of cytidine triphosphate(CTP) (CTP) only is 88%, the transformation efficiency of uridine triphosphate (UTP) only is 85%, and the reaction times reaches 7 hours, yeast can not use repeatedly, and product separation also has certain difficulty.
The objective of the invention is to disclose a kind of carrageenin-konjac polysaccharide gum blend fixed microorganism carrier, and the application of this carrier in preparation ribonucleoside triphosphote such as adenosine triphosphate (ATP), black guanosine triphosphate (GTP), cytidine triphosphate(CTP) (CTP) and uridine triphosphate production processes such as (UTP).
Design of the present invention is such:
The contriver is at independent when using carrageeenen immobilized microbe, it is low to exist gel-strength, fragility and bleeding, defectives such as the easy fragmentation of particle, the gum blend that adopts carrageenin sodium salt and konjac polysaccharide is as Immohilized microorganism carrier, microorganism is embedded in the gum blend of carrageenin sodium salt and konjac polysaccharide, forms a kind of carrageenin-konjac polysaccharide gum blend immobilized microorganism.The main component of konjac polysaccharide is a glucomannan, it be D-glucose and D-mannosans by a certain percentage, with β-1,4 glycosidic link bonded complex polysaccharides, evidence: carrageenin-konjac polysaccharide gum blend immobilized microorganism not only has the advantage of gelatin glutaraldehyde immobilized microorganism, and the shortcoming can avoid using carrageenin separately the time.
The said carragheen of the present invention-SKGM compound gel fixed microorganism carrier is a composition, and its component and content are as follows: the preferred ratio of kappa-carrageenan sodium salt 2～10% SKGMs, 0.4～4% water surplus is: the ratio of kappa-carrageenan sodium salt 3～6% SKGMs 0.6～1.8% water surplus the best is: the equal weight percentage of content of kappa-carrageenan sodium salt 3.2% SKGM 0.8% water surplus said components.
The preparation process of said kappa-carrageenan-konjac polysaccharide gum blend fixed microorganism carrier is foolproof, kappa-carrageenan sodium salt and the smart powder of konjac polysaccharide are dissolved in the water according to the above ratio, can obtain said carrier, this carrier can be incubated standby under 40～50 ℃ temperature.
Said kappa-carrageenan-konjac polysaccharide gum blend fixed microorganism carrier can be used for the various microorganisms of embedding, as yeast, brevibacterium flavum, Bacillus subtilus or intestinal bacteria etc., thereby obtains kappa-carrageenan-konjac polysaccharide gum blend immobilized microorganism.Its preparation process mainly may further comprise the steps:
(1) with microorganism (water content 80%, viable count 90%) with physiological saline washing, and under 30～40 ℃ temperature, is incubated 20～40 fens, pours into then in the mixture of the smart powder of carrageenin sodium salt konjac polysaccharide of above-mentioned heat, stir, pour the film that is paved into 2～4mm in the dish into.The add-on of microorganism is 45～55% of a gross weight;
(2) adding concentration is peptizer-KCl solution of 0.3%～2% (wt%), add-on be contain microorganism amount of the mixture 0.15～1%, place 4～20 ℃ environment again, it is solidified, it is evenly cut into fritter;
(3) fritter is dropped in the peptizer that concentration is 0.3%～2% (wt%)-KCl solution and soak, it is fully solidified, and then drop into to soak in the glutaraldehyde solution that concentration is 0.5%～2% (wt%) and carried out crosslinked reinforcing in 20～60 minutes, can obtain carrageenin-konjac polysaccharide gum blend immobilized microorganism, be placed on preserve in-15～-30 ℃ the environment standby;
When microorganism is yeast, then can be used for preparing ribonucleoside triphosphote, as adenosine triphosphate (ATP), black guanosine triphosphate (GTP), cytidine triphosphate(CTP) (CTP) and uridine triphosphate (UTP).Preparation process is as follows:
With said carrageenin-konjac polysaccharide gum blend fixed yeast and reaction solution by placing reactor, the concussion reaction, after reaction finished, with reaction product centrifuging, filtrate was handled with 717 anionite-exchange resin, absorption ribonucleoside triphosphote wherein, use (pH=2) NaCl wash-out of 0.7mol/L then, collect elutriant, add ethanol, make the ribonucleoside triphosphote precipitation, can get target product-ribonucleoside triphosphote after filtration with after the vacuum-drying.Fixed yeast then can be reused once more.
Said reaction solution is a composition, and its mole ratio of components is: substrate 20～80 mmol/L glucose 100～300 mmol/L sal epsom, 4～20 mmol/L phosphoric acid buffers, 150～350 mmol/L (pH=6.5～7.5) nadide 0～3 mmol/L water surplus;
Carrageenin-konjac polysaccharide gum blend fixed yeast is 1 (gram) with the ratio of reaction solution: (0.7～1.5) (ml);
Preferred enzymatic reaction temperature is 26～38 ℃, and best temperature is 36 ℃, and the reaction times is 1～4 hour, and the preferred reaction times is 2～3 hours.
Said substrate is a kind of in adenosine (AR), adenylic acid (AMP) (AMP), black thuja acid (GMP), cytidylic acid (CMP) or the uridylic acid (UMP);
When substrate is adenosine (AR) or adenylic acid (AMP) (AMP), can obtain adenosine triphosphate (ATP); When substrate is black thuja acid (GMP), can obtain black guanosine triphosphate (GTP); When substrate is cytidylic acid (CMP), can obtain cytidine triphosphate(CTP) (CTP); When substrate is uridylic acid (UMP), can obtain uridine triphosphate (UTP).
Said nadide is a VITAMIN B4 acid amides dinucleotides.
Below with reference to embodiment the said content of the present invention is further described.
Embodiment 1
8 gram kappa-carrageenan sodium salts and the smart powder of 2 gram konjac polysaccharide are dissolved in the 250ml water, insulation is standby under 50 ℃ temperature, cereuisiae fermentum (water content 80% with 250 grams, viable count 90%) washs with physiological saline, and under 36 ℃ temperature, be incubated 30 minutes, pour into then in the mixture of the smart powder of carrageenin sodium salt konjac polysaccharide of above-mentioned heat, stir, pour the film that is paved into 2mm in the dish into, adding 500ml concentration is the KCl solution of 0.3% (wt%), place 4 ℃ environment again, it is solidified, it is evenly cut into to drop into behind the fritter in the KCl solution that concentration is 0.3% (wt%) soak, it is fully solidified, and then drop in the glutaraldehyde solution that concentration is 0.5% (wt%) and soaked 30 minutes, carry out crosslinked reinforcing, can obtain carrageenin-konjac polysaccharide gum blend fixed yeast, be placed on preserve in-20 ℃ the environment standby.After measured, its every mechanical property all is better than similar carrier, and the result is as follows:
| Carrageenin | Sodium alginate | Gum blend | |
| Gel density (g/cm 2) | 1008 | 1530 | 2020 |
| Elasticity | + | +++ | +++ |
Embodiment 2
Take by weighing embodiment 1 prepared kappa-carrageenan-konjac polysaccharide gum blend fixed yeast 15 grams and place reaction flask, add AR 40mmol, glucose 250mmol, sal epsom 8mmol, phosphoric acid buffer 250mmol (pH=6.5～7.5), nadide 1mmol shakes reaction 2 hours down at 36 ℃, with HPCC method or electrophoretic method reaction solution is analyzed, the transformation efficiency of product-ATP is 99%.After reaction finishes, with reaction product centrifuging, filtrate is handled with 717 anionite-exchange resin, absorption ribonucleoside triphosphote wherein, use (pH2) NaCl wash-out of 0.7mol/L then, collect elutriant, add ethanol, make the ribonucleoside triphosphote precipitation, can get target product-ribonucleoside triphosphote after filtration with after the vacuum-drying.
Embodiment 3～6
Operating process is identical with embodiment 2, and substrate is replaced with AMP, GMP, CMP and VMP respectively, and its product is respectively ATP, GTP, CTP and UTP, and its transformation efficiency is respectively 90%, 96%, 98% and 98%.
By above-mentioned disclosed technical scheme and embodiment as seen, advantage of the present invention is fairly obvious:
(1) immobilized microorganism intensity height, good springiness is difficult for fragmentation, helps prolonged and repeated use;
(2) kappa-carrageenan-konjac polysaccharide gum blend is difficult for by microbiological degradation;
(3) the substrate conversion efficiency height reaches as high as 99%;
(4) immobilized microorganism can be repeatedly used, and production cost is low.
(5) immobilized microorganism and product separate easily, microorganism is deposited in the embedded particles, does not bring in the reaction soln, has simplified separation circuit, and can obtain high-quality product.
Claims (8)
1. carrageenin-konjac polysaccharide gum blend fixed microorganism carrier is characterized in that component and content are:
Kappa-carrageenan sodium salt 2～10%
Konjac polysaccharide 0.4～4%
Water 86～97.6%
The equal weight per-cent of the content of said components.
2. carrageenin as claimed in claim 1-konjac polysaccharide gum blend fixed microorganism carrier is characterized in that component and content are:
Kappa-carrageenan sodium salt 3～6%
Konjac polysaccharide 0.6～1.8%
Water 92.2～96.4%
The equal weight per-cent of the content of said components.
3. carrageenin as claimed in claim 2-konjac polysaccharide gum blend fixed microorganism carrier is characterized in that component and content are:
Kappa-carrageenan sodium salt 3.2%
Konjac polysaccharide 0.8%
Water 96%
The equal weight per-cent of the content of said components.
4. the application of carrageenin as claimed in claim 1-konjac polysaccharide gum blend fixed microorganism carrier is characterized in that: can be used for preparing ribonucleoside triphosphote.
5. the application of carrageenin as claimed in claim 2-konjac polysaccharide gum blend fixed microorganism carrier is characterized in that: can be used for preparing ribonucleoside triphosphote.
6. the application of carrageenin as claimed in claim 3-konjac polysaccharide gum blend fixed microorganism carrier is characterized in that: can be used for preparing ribonucleoside triphosphote.
7. as the application of claim 4 or 5 or 6 described carrageenins-konjac polysaccharide gum blend fixed microorganism carrier, it is characterized in that: 1. earlier with conventional method carrier embedding yeast, obtain carrageenin-konjac polysaccharide gum blend fixed yeast, 2. and then with reaction solution react, 3. isolate ribonucleoside triphosphote with conventional method at last from reaction solution, its processing condition are: the mole of (1) reaction solution consists of:
Substrate 20～80 mmol/L
Glucose 100～300 mmol/L
Sal epsom 4～20 mmol/L
Phosphoric acid buffer 150～350 mmol/L (pH=6.5～7.5)
Nadide 0～3 mmol/L
1 liter in water; (2) carrageenin-konjac polysaccharide gum blend fixed yeast is 1 (gram) with the ratio of reaction solution: (0.7～1.5) (ml); (3). the enzymatic reaction temperature is 26～38 ℃, and the reaction times is 1～4 hour;
Said substrate is a kind of in adenosine (AR), adenylic acid (AMP) (AMP), black thuja acid (GMP), cytidylic acid (CMP) or the uridylic acid (UMP);
Said nadide is a VITAMIN B4 acid amides dinucleotides.
8. as the application of claim 1 or 2 or 3 described carrageenins-konjac polysaccharide gum blend fixed microorganism carrier, it is characterized in that: can be used for preparing immobilized microorganism, said microorganism is yeast, brevibacterium flavum, Bacillus subtilus or intestinal bacteria.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99113707A CN1086736C (en) | 1999-05-17 | 1999-05-17 | Microorganism carrier fixed with glue compounded with carragheen and konjak polysaccharide and use thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN99113707A CN1086736C (en) | 1999-05-17 | 1999-05-17 | Microorganism carrier fixed with glue compounded with carragheen and konjak polysaccharide and use thereof |
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| Publication Number | Publication Date |
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| CN1234444A CN1234444A (en) | 1999-11-10 |
| CN1086736C true CN1086736C (en) | 2002-06-26 |
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| CN99113707A Expired - Fee Related CN1086736C (en) | 1999-05-17 | 1999-05-17 | Microorganism carrier fixed with glue compounded with carragheen and konjak polysaccharide and use thereof |
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Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100379859C (en) * | 2005-03-04 | 2008-04-09 | 上海秋之友生物科技有限公司 | Immobilized nucleoside phosphorylase microbe and its application in synthesizing purine nucleosidase |
| CN100363376C (en) * | 2006-06-12 | 2008-01-23 | 南京工业大学 | Crystallization process of 5'-nucleoside sodium triphosphate |
| CN100582228C (en) * | 2007-03-29 | 2010-01-20 | 中国农业科学院农产品加工研究所 | Immobilization lipase, and preparation method |
| CN101157914B (en) * | 2007-09-20 | 2011-06-15 | 西北师范大学 | Bentonite, biological polysaccharide plural gel immobilized cell carrier and preparation method thereof |
| CN101265453B (en) * | 2008-04-18 | 2010-11-03 | 中国药科大学 | Beer yeast strain containing CMP kinase and CDP kinase and application |
| CN105922783A (en) * | 2016-06-30 | 2016-09-07 | 合肥环照高分子材料厂 | High-moisture-retention fountain solution compounded by k-carrageenan and konjac glucomannan and preparation method thereof |
| CN106113964A (en) * | 2016-06-30 | 2016-11-16 | 合肥环照高分子材料厂 | A kind of high antioxidant fountain solution compounding containing κ carrageenan Konjac glucomannan and preparation method thereof |
| CN106142879A (en) * | 2016-06-30 | 2016-11-23 | 合肥环照高分子材料厂 | A kind of height hard water resistance property fountain solution compounding containing κ carrageenan Konjac glucomannan and preparation method thereof |
| CN105966104A (en) * | 2016-06-30 | 2016-09-28 | 合肥环照高分子材料厂 | Insect-preventing fountain solution containing kappa-carrageenan-amorophophallus konjac compound and preparation method of insect-preventing fountain solution |
| CN106113968A (en) * | 2016-07-19 | 2016-11-16 | 合肥鼎亮光学科技有限公司 | A kind of UV quick-drying hectograph fountain solution compounding containing κ carrageenan Konjac glucomannan and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86102156A (en) * | 1986-03-28 | 1987-10-14 | 江苏省食品发酵研究所 | The novel vector that fixation of microbe cell and enzyme are used |
| CN1038124A (en) * | 1988-05-31 | 1989-12-20 | 大连轻工业学院 | The natural carrier of fixation of microbe cell and enzyme and device thereof |
| CN1072957A (en) * | 1992-12-18 | 1993-06-09 | 华东化工学院 | A kind of fixed yeast cell and the application on adenosine triphosphate is produced thereof |
-
1999
- 1999-05-17 CN CN99113707A patent/CN1086736C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86102156A (en) * | 1986-03-28 | 1987-10-14 | 江苏省食品发酵研究所 | The novel vector that fixation of microbe cell and enzyme are used |
| CN1038124A (en) * | 1988-05-31 | 1989-12-20 | 大连轻工业学院 | The natural carrier of fixation of microbe cell and enzyme and device thereof |
| CN1072957A (en) * | 1992-12-18 | 1993-06-09 | 华东化工学院 | A kind of fixed yeast cell and the application on adenosine triphosphate is produced thereof |
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| CN1234444A (en) | 1999-11-10 |
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