CN1966703A - Process for one-step fermentation production of gulonic acid (VC precursor) from glucose - Google Patents
Process for one-step fermentation production of gulonic acid (VC precursor) from glucose Download PDFInfo
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- CN1966703A CN1966703A CN 200610144108 CN200610144108A CN1966703A CN 1966703 A CN1966703 A CN 1966703A CN 200610144108 CN200610144108 CN 200610144108 CN 200610144108 A CN200610144108 A CN 200610144108A CN 1966703 A CN1966703 A CN 1966703A
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- glucose
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- xanthomonas campestris
- shines
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Abstract
The invention discloses a method for producing gulonic acid (VC precursor). The method comprises the steps as follows: 1) inducing proper amount of Xanthomonas campestris with ray, 2) culturing on a plate at 25-35DEG C for 2-5d, selecting the fine colony which satisfies the requirement of smooth colony, inoculating in a conical flask with culture fluid for culturing, 3) adding into the medium containing glucose for culturing to producegulonic acid. The invention realizes direct production ofgulonic acid from glucose to produce VC, with a converting ratio of 35-50%, has the advantages of simple production, reduces the production cost greatly.
Description
Technical field
The present invention relates to a kind of ascorbic preparation technology.
Background technology
Vitamins C (abbreviating VC as) is that human body one of can not self synthetic VITAMIN, participates in multiple metabolic process in the body, can alleviate the fragility of capillary vessel, increases Abwehrkraft des Koepers.Be used to prevent vitamin C deficiency, also can be used for the assisting therapy of various chronic infectious diseases and purpura etc.Can stop the generation of carcinogenic substance nitrous acid amine.VC is good antioxidant, and food is played freshening effect, has a wide range of applications at food, beverage and feedstuff industry, and demand is along with the people's standard of living improves constantly increase, and ascorbic global demand amount was more than 12 ten thousand tons in 2005.Ascorbic production preparation mainly comprises following several:
1) Lai Shi method
Vitamins C extracted from citric acid and gets as far back as 1910～nineteen twenty-one, and output is few, cost an arm and a leg, and can not satisfy the demand far away.The Lai Shi (Reichstein) of fritz's invention in 1933 etc. succeeds with chemical process synthesise vitamins C, and pushes to industrial production, and the vitamins C price is descended significantly, and price drops to over one hundred unit dollar from more than 2000 dollar of per kilogram.The Lai Shi chemical method need pass through five processes (one-step fermentation, ketonize, oxidation, conversion and refining), operate continuously is had any problem, not only with the output of a large amount of toxic gases and " three wastes ", and production environment there is strict fire-proof and explosion-proof safety requirements in the production.Constantly enlarge along with synthesis technique constantly improves with industrial scale, price descends year by year, though development therebetween has many can not be better than the Lai Shi method in technology and price for the synthetic route that adopts.Therefore at present in the world still with Lai Shi method and the production of improvement route thereof.
The Lai Shi method is to be raw material with D-glucose; be hydrogenated to sorbyl alcohol earlier; with the acetic bacteria fermentation is sorbose; again with the acetone ketonize protect diacetone sorbose; change into the ancient dragon acid of diacetone-2-ketone-L-through chemical oxygen; be hydrolyzed to the ancient dragon acid of 2-ketone-L-, transform with acid or alkali again, get L-xitix (vitamins C).
Produce the production technique of diacetone sorbose by sorbyl alcohol, sorbose from glucose, basic identical both at home and abroad, but it is then different to be oxidized to the ancient imperial acid of acetone-2-ketone-L-from diacetone sorbose, adopt the potassium permanganate method as Japan, China and U.S. Roche all use sodium-hypochlorite process, and U.S. Fa Zeer then adopts palladium catalyst method.The ancient imperial acid of 2-ketone-L-after the ancient imperial acid hydrolysis of diacetone-2-ketone-L-can be adopted acid to transform and generate vitamins C.
The advantage of this route is yield height, process stabilizing, quality of finished product good.Shortcoming is that reactions steps is many, the consumption raw material is many (needing a large amount of acetone, sulfuric acid and liquid caustic soda etc.).Existing domestic seldom employing.
For shortening reactions steps; imagination is protected without acetone from sorbose, becomes the ancient dragon acid of 2-ketone-L-with chemical direct oxidation, though the past is through exploring in many ways; but yield is all low; still continue research in recent years, making catalyst as Japan's report with Pt and Pd and Pb or Bi, in the solution that contains 8% sorbose; pH remains on 7～8; the bubbling air oxidation, yield can reach 87%, increases than in the past.
Owing to the three-step reaction technical maturity of producing diacetone sorbose from glucose, yield is all higher, does not therefore have much progress in recent years.
From diacetone-l-sorbose to diacetone-oxidizing reaction of the ancient dragon acid of 2-ketone-L-and the ancient dragon acid of 2-ketone-L-is to ascorbic conversion reaction, then in continuous research.
2) two-step fermenting
China scientist researchs and produces ascorbic novel method the seventies and succeeds, and this method was that the ancient dragon acid of ketone-L-was continuously fermented by two steps from sorbyl alcohol to 2-, had pushed to industrial production in 1970.Two-step fermenting is actually simplifies and has shortened the Lai Shi route, though this method industrial production is historical short, but progressively demonstrate its superiority, the false pseudomonas bacillus of one a strain oxidizing glucose acidfast bacilli (" little bacterium ") and a strain (" big bacterium "), the hybrid bacterial strain that two kinds of bacterium of size make up naturally carry out the fermentation of second step again and generate the ancient dragon acid of (-) 2-ketone group-L-.Particularly with bio-oxidation replace chemical oxidation, slice off the Lai Shi method the ketonize hydrolysis reaction, saved that 80% acetone and other chemical industry raw material, work simplification, raw material consumption are low, raw materials cost and factory cost all be lower than the Lai Shi method, three-waste pollution is few, equipment corrosion is little, helps safety in production.
However, the manufacturing cost of this method is still very high with respect to the VC international market price, and special formula sorbyl alcohol is wherein produced 1 ton of vitamins C and will be dropped into 1.7 tons of sorbyl alcohols, makes to adopt this method to produce the manufacturer of VC almost without any profit.China's output accounts for 68% of the total production in the world.World market vitamins C price drops many times, and the profit of manufacturer production is fewer and feweri, change such situation, and unique way is exactly to reduce production costs to improve profit again.
Summary of the invention
At the problem of above-mentioned existence, the object of the present invention is to provide a kind of method of producing ancient imperial sour (VC precursor) through one-step fermentation by glucose, this method can reduce production costs greatly, and manufacture method is more simple.
For achieving the above object, a kind of method of producing ancient dragon acid (VC precursor) through one-step fermentation by glucose provided by the invention comprises the steps:
1) chooses an amount of xanthomonas campestris (Xanthomonas campestris) and carry out mutagenic exposure with ray;
2) postradiation xanthomonas campestris after 2～5 days, is picked out bacterium colony preferably with the plate culture medium cultivation earlier under 25～35 ℃ of conditions, promptly satisfies the smooth smooth requirement of bacterium colony, is seeded in the Erlenmeyer flask that contains nutrient solution and cultivates;
3) cultivate on 30～40 ℃ of shaking tables adding to contain in the dextrose culture-medium, thereby prepare ancient dragon acid through the Zymomonas mobilis after mutagenesis and the cultivation.
Further, the radiation modality of described step 1) can be selected for use
60Co shines 200～3200Gy, shines 1.5～24 minutes, or with UV-lamp 15～20W, apart from 20～50 centimeters, shines for 10～35 seconds.
Further, described nutrient solution is that every liter of solution contains glucose 35～45g, yeast extract 1～5g, K
2HSO
42～8g and MgSO
47H
2The aqueous solution of O 0.6g.
The present invention has realized directly preparing the ancient dragon acid that is used to produce VC from glucose by cultivating suitable bacterial strain, transformation efficiency can reach 35～50%, not only manufacture method is simple, and greatly reduces production cost, and one ton of sour comparable two step method of ancient dragon of every production is saved 6000～10000 yuan.
Embodiment
Xanthomonas campestris is mainly used in the production xanthan gum, and xanthan gum is good suspension agent and an emulsion stabilizer in the foodstuff additive.The present invention utilizes xanthomonas campestris to go out ancient dragon acid through direct production after the specific culture, belongs to a kind of new purposes invention of xanthomonas campestris.Below by embodiment the present invention is done detailed explanation.
Embodiment 1
Specifically according to following steps:
1) choosing an amount of xanthomonas campestris adopts
60Co shines 200Gy, shines 24 minutes;
2) postradiation xanthomonas campestris is under 35 ℃ of conditions, after cultivating 2～5 days with plate culture medium earlier, pick out bacterium colony preferably, promptly satisfy the smooth smooth requirement of bacterium colony, be seeded in then in the Erlenmeyer flask that contains the 50ml nutrient solution and cultivate, nutrient solution is that every liter of solution contains glucose 35g, yeast extract 5g, K
2HSO
48g and MgSO
47H
2The aqueous solution of O 0.6g;
3) cultivate on 40 ℃ of shaking tables adding to contain in the dextrose culture-medium, thereby prepare ancient dragon acid through the Zymomonas mobilis after mutagenesis and the cultivation.
Adopt the ancient imperial sour transformation efficiency of this method preparation can reach 35%.
Embodiment 2
Specifically according to following steps:
1) choosing an amount of xanthomonas campestris adopts
60Co shines 3200Gy, shines 1.5 minutes;
2) postradiation xanthomonas campestris is under 30 ℃ of conditions, after cultivating 2～5 days with plate culture medium earlier, pick out bacterium colony preferably, promptly satisfy the smooth smooth requirement of bacterium colony, be seeded in the Erlenmeyer flask that contains the 50ml nutrient solution and cultivate, nutrient solution is that every liter of solution contains glucose 40g, yeast extract 3g, K
2HSO
45g and MgSO
47H
2The aqueous solution of O 0.6g;
3) cultivate on 30 ℃ of shaking tables adding to contain in the dextrose culture-medium, thereby prepare ancient dragon acid through the Zymomonas mobilis after mutagenesis and the cultivation.
Adopt the ancient imperial sour transformation efficiency of this method preparation can reach 40%.
Embodiment 3
Specifically according to following steps:
1) choosing an amount of xanthomonas campestris adopts
60Co shone 15 minutes with 1800Gy;
2) postradiation xanthomonas campestris is under 25 ℃ of conditions, after cultivating 2～5 days with plate culture medium earlier, pick out bacterium colony preferably, promptly satisfy the smooth smooth requirement of bacterium colony, be seeded in the Erlenmeyer flask that contains the 50ml nutrient solution and cultivate, nutrient solution is that every liter of solution contains glucose 45g, yeast extract 1g, K
2HSO
42g and MgSO
47H
2The aqueous solution of O 0.6g;
3) cultivate on 37 ℃ of shaking tables adding to contain in the dextrose culture-medium, thereby prepare ancient dragon acid through the Zymomonas mobilis after mutagenesis and the cultivation.
Adopt the ancient imperial sour transformation efficiency of this method preparation can reach 50%.
Embodiment 4
Specifically according to following steps:
1) choose an amount of xanthomonas campestris, at first be inoculated into and contain in the dextrose culture-medium, 25 ℃ 16 hours, centrifugal (10,000 γ/minute) washed several times with centrifugal xanthomonas campestris body and function sterile distilled water after 5 minutes, made 10
6-8Individual/the milliliter bacteria suspension, get 2 milliliters of bacteria suspensions and join in 5 centimeters culture dish, adopt UV-lamp 15～20W, 20～50 centimeters of distances shone for 10～35 seconds;
2) postradiation xanthomonas campestris is under 20～30 ℃ of conditions, after cultivating 2～5 days with plate culture medium earlier, pick out bacterium colony preferably, promptly satisfy the smooth smooth requirement of bacterium colony, be seeded in the Erlenmeyer flask that contains the 50ml nutrient solution and cultivate, nutrient solution is that every liter of solution contains glucose 45g, yeast extract 1g, K
2HSO
42g and MgSO
47H
2The aqueous solution of O 0.6g (under 610～700 millimicrons of wavelength light, operating);
3) cultivate on 37 ℃ of shaking tables adding to contain in the dextrose culture-medium, thereby prepare ancient dragon acid through the Zymomonas mobilis after mutagenesis and the cultivation.
Adopt the ancient imperial sour transformation efficiency of this method preparation can reach 50%.
Though the foregoing description just passes through
60The irradiation of Co and UV-lamp is illustrated, but the present invention is not limited thereto, can also adopt X-ray irradiation, laser radiation or other can be with xanthomonas campestris with methods such as ray mutagenesis, and these methods also all belong to the scope of protection of the invention.
Claims (3)
1, a kind of method of producing ancient imperial sour (VC precursor) through one-step fermentation by glucose is characterized in that, comprises the steps:
1) chooses an amount of xanthomonas campestris and carry out mutagenic exposure with ray;
2) postradiation xanthomonas campestris after 2～5 days, is picked out bacterium colony preferably with the plate culture medium cultivation earlier under 25～35 ℃ of conditions, promptly satisfies the smooth smooth requirement of bacterium colony, is seeded in the Erlenmeyer flask that contains nutrient solution and cultivates;
3) cultivate on 30～40 ℃ of shaking tables adding to contain in the dextrose culture-medium, thereby prepare ancient dragon acid through the Zymomonas mobilis after mutagenesis and the cultivation.
2, according to claim 1 method of producing ancient imperial sour (VC precursor) through one-step fermentation, it is characterized in that the radiation modality of described step 1) can be selected for use by glucose
60Co shines 200～3200Gy, shines 1.5～24 minutes, or with UV-lamp 15～20W, apart from 20～50 centimeters, shines for 10～35 seconds.
3, according to claim 1 or 2 one kind of method of producing ancient imperial sour (VC precursor) through one-step fermentation, it is characterized in that described nutrient solution is that every liter of solution contains glucose 35～45g, yeast extract 1～5g, K by glucose
2HSO
42～8g and MgSO
47H
2The aqueous solution of O 0.6g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610144108 CN1966703A (en) | 2006-11-27 | 2006-11-27 | Process for one-step fermentation production of gulonic acid (VC precursor) from glucose |
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|---|---|---|---|
| CN 200610144108 CN1966703A (en) | 2006-11-27 | 2006-11-27 | Process for one-step fermentation production of gulonic acid (VC precursor) from glucose |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105601050A (en) * | 2016-03-17 | 2016-05-25 | 富阳鸿祥技术服务有限公司 | Biological fermentation-heavy metal sedimentation purifying method |
| CN105621632A (en) * | 2016-03-17 | 2016-06-01 | 富阳鸿祥技术服务有限公司 | Biological fermentation and heavy metal precipitation purification pool |
| CN110938564A (en) * | 2019-12-05 | 2020-03-31 | 石药集团维生药业（石家庄）有限公司 | Method for promoting growth and metabolism of ketogenic gulonospora |
-
2006
- 2006-11-27 CN CN 200610144108 patent/CN1966703A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105601050A (en) * | 2016-03-17 | 2016-05-25 | 富阳鸿祥技术服务有限公司 | Biological fermentation-heavy metal sedimentation purifying method |
| CN105621632A (en) * | 2016-03-17 | 2016-06-01 | 富阳鸿祥技术服务有限公司 | Biological fermentation and heavy metal precipitation purification pool |
| CN110938564A (en) * | 2019-12-05 | 2020-03-31 | 石药集团维生药业（石家庄）有限公司 | Method for promoting growth and metabolism of ketogenic gulonospora |
| CN110938564B (en) * | 2019-12-05 | 2023-04-14 | 石药集团维生药业（石家庄）有限公司 | Method for promoting growth and metabolism of ketogenic gulonospora |
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Open date: 20070523 |