CN1282749C - Course feeding fermentation method for preparing astaxanthin with molasses or starch sugar as materials - Google Patents
Course feeding fermentation method for preparing astaxanthin with molasses or starch sugar as materials Download PDFInfo
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- CN1282749C CN1282749C CN 200310112328 CN200310112328A CN1282749C CN 1282749 C CN1282749 C CN 1282749C CN 200310112328 CN200310112328 CN 200310112328 CN 200310112328 A CN200310112328 A CN 200310112328A CN 1282749 C CN1282749 C CN 1282749C
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- astaxanthin
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- dian fentang
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Abstract
The present invention relates to a course feeding fermentation method for preparing astaxanthin with molasses or starch sugar as materials. Microorganisms are inoculated to a liquid culture medium, the dissolved oxygen concentration is controlled from 10 to 80% at a temperature of 18 to 22 DEG C, and the pH in a cultivation process is from 3 to 6. When the concentration of remnant sugar in the culture medium is lower than 15 g/L, molasses or starch sugar is added until the sugar concentration of the culture medium reaches 10 to 60 g/L for continuous cultivation; the cultivation is ended after 72 to 120 hours. Red phaffia rhodozyma is used for inoculating microorganisms, and the culture medium is molasses or starch sugar in which an inorganic or organic nitrogen source is added. The high-density culture is obtained through feeding cultivation in the present invention; thus, the biomass, the output of astaxanthin and the yield in unit time and the unit volume of devices are enhanced, and the production cost is reduced.
Description
Technical field
The present invention relates to biotechnology and technical field, particularly a kind of method of utilizing molasses or the middle fed-batch fermentation of Dian Fentang raw material to produce astaxanthin.
Technical background
Astaxanthin is the main pigment of Crustacean, salmon and various birds, and its antioxidant property than the strong 100-500 of vitamin-E doubly, and is stronger 10 times than carotene; Animal experiment shows that astaxanthin has antitumor, anti-ultraviolet radiation, raise immunity, is the powerful antioxidant of hydrocoles and the inhibitor of lipid peroxidation; Ophthalmic diseases, cardiovascular disorder and rheumatoid arthritis also there is mitigation.Current research finds that astaxanthin helps to eliminate the physical discomfort that produces because of the time difference, and its effect is also more obvious than melatonin commonly used.To the daily schedule fall into the students in middle and primary schools of what disorder and old man recover normal rhythm of life may also helpful effect.Astaxanthin has using value in the following aspects: the 1. good tinting material of aquatic products and livestock culture, improve the product of breed surviving rate, rate of body weight gain and feed conversion rate simultaneously; 2. foodstuff additive (tinting material and antioxidant); 3. healthcare products and medicine (raise immunity and prevent lipid peroxidation etc.); 4. makeup (anti-ultraviolet radiation and anti-oxidant etc.).Prior art mainly adopts the method for producing astaxanthin of intermittently cultivating, and the biomass of acquisition and astaxanthin yield are all lower,
Summary of the invention
The object of the present invention is to provide a kind of is that the method for producing astaxanthin was cultivated in feed supplement in the middle of main raw material carried out with molasses or Dian Fentang, cultivate by feed supplement, reach high-density culture, thereby improve the unit time unit volume yield of biomass and astaxanthin yield and equipment, reduce production costs.
A kind of method of utilizing molasses or the middle fed-batch fermentation of Dian Fentang raw material to produce astaxanthin of the present invention, be in liquid nutrient medium with microbial inoculant, under 18-22 ℃ temperature, dissolved oxygen concentration is controlled at 10-80%, the pH of culturing process is 3-6, when remaining sugar concentration in the substratum is lower than 15 grams per liters, add molasses or Dian Fentang to substratum sugar concentration 10-60 grams per liter and continue to cultivate, after 72-120 hour, finish to cultivate; The microorganism that is used to inoculate is red Fife's yeast, and substratum is that molasses or Dian Fentang add inorganic or organic nitrogen source.
Described red Fife's yeast is As2.1557 or its mutant strain.
Described carbon source is molasses or Dian Fentang.
Described inorganic nitrogen-sourced be ammonium sulfate, ammonium nitrate, ammonium chloride or urea.
Described organic nitrogen source is yeast powder or yeast extract paste, peptone, corn steep liquor.
The present invention can be with shaking bottle or using bioreactor culture, and when using bioreactor culture, dissolved oxygen concentration preferably is controlled at 10-80%.
Described culturing process can be added ethanol, and consumption is 0-8g/L.
Phaffiafhodozyma has fast growth, and fermentation period is short, the content astaxanthin height, and thalline can be the good resource of astaxanthin directly as advantages such as feeds.Prior art adopts biomass and the astaxanthin yields of intermittently cultivating many acquisitions all lower, adopt phaffiafhodozyma to produce astaxanthin, cultivate by feed supplement, can improve the unit time unit volume yield of biomass and astaxanthin yield and equipment, thus reduce production cost.
The method of utilizing molasses or the middle fed-batch fermentation of Dian Fentang raw material to produce astaxanthin of the present invention, its advantage is: (1) fermention medium is very simple, have only molasses or Dian Fentang and inorganic nitrogen-sourced or organic nitrogen source, as ammonium sulfate, urea, yeast powder, corn steep liquor etc.; (2) need not add any other material, the cell well-grown more approaches suitability for industrialized production.
Embodiment
Embodiment 1
The bacterial strain that the bacterial classification that adopts: As2.1557 obtains through mutagenesis and fusion.
Shake bottle batch culture method: have only 20 grams per liter molasses and 5 grams per liter ammonium sulfate in the fermention medium, transfer pH 5.0,18-22 ℃, cultivated 72 hours for 160 rev/mins, collect thalline.Dry cell weight 7.25 grams per liters, cell content astaxanthin 812 micrograms/gram stem cell, astaxanthin yield reaches 5887 micrograms per litre.
Embodiment 2
Other conditions when remaining sugar concentration in the nutrient solution is lower than 15 grams per liters, are added 20 grams per liter molasses with example 1, continue to be cultured to 84 hours, collect thalline.Dry cell weight 13.25 grams per liters, cell content astaxanthin 786 micrograms/gram stem cell, astaxanthin yield reaches 10415 micrograms per litre.
Embodiment 3
Other conditions are with example 1, and carbon source adopts 20 grams per liter Dian Fentangs.Dry cell weight 6.86 grams per liters, cell content astaxanthin 892 micrograms/gram stem cell, astaxanthin yield reaches 6119 micrograms per litre
Embodiment 4
Other conditions are with example 1, and nitrogenous source adopts 5 grams per liter ammonium chlorides.Dry cell weight 7.08 grams per liters, cell content astaxanthin 796 micrograms/gram stem cell, astaxanthin yield reaches 5636 micrograms per litre.
Embodiment 5
Other conditions are with example 1, and nitrogenous source adopts 5 grams per liter ammonium nitrate.Dry cell weight 6.76 grams per liters, cell content astaxanthin 782 micrograms/gram stem cell, astaxanthin yield reaches 5286 micrograms per litre.
Embodiment 6
Other conditions are with example 1, and nitrogenous source adopts 5 grams per liter urea.Dry cell weight 4.76 grams per liters, cell content astaxanthin 912 micrograms/gram stem cell, astaxanthin yield reaches 4341 micrograms per litre.
Embodiment 7
Other conditions are with example 1, and nitrogenous source adopts 5 grams per liter yeast powders.Dry cell weight 9.48 grams per liters, cell content astaxanthin 946 micrograms/gram stem cell, astaxanthin yield reaches 8968 micrograms per litre.
Embodiment 8
Other conditions are with example 1, and nitrogenous source adopts 5 grams per liter peptones.Dry cell weight 8.12 grams per liters, cell content astaxanthin 786 micrograms/gram stem cell, astaxanthin yield reaches 6382 micrograms per litre.
Embodiment 9
Other conditions are with example 1, and nitrogenous source adopts 5 grams per liter corn steep liquors.Dry cell weight 7.92 grams per liters, cell content astaxanthin 754 micrograms/gram stem cell, astaxanthin yield reaches 5972 micrograms per litre.
Embodiment 10
Other conditions are transferred pH 6.0 with example 1.Dry cell weight 7.13 grams per liters, cell content astaxanthin 804 micrograms/gram stem cell, astaxanthin yield reaches 5732 micrograms per litre.
Embodiment 11
Other conditions are transferred pH 3.0 with example 1.Dry cell weight 6.21 grams per liters, cell content astaxanthin 764 micrograms/gram stem cell, astaxanthin yield reaches 4744 micrograms per litre.
Embodiment 12
Bio-reactor is criticized culture method: the amplification of embodiment 1 is that (New BrunswickScientific, Bioflo3000 USA) carry out in the bio-reactor, and liquid amount is 3.5 liters at 5 liters.Initial Ventilation Rate is 1.75 liters/minute, and initial stir speed (S.S.) is 150 rev/mins, and inoculum size is 10%, with 10% sulfuric acid and 10% sodium hydroxide control pH 5.0.Other fermentation conditions are with embodiment 1.Along with the growth of cell, oxygen consumption rate increases gradually, in order to keep dissolved oxygen level 40%, increases Ventilation Rate and stir speed (S.S.) gradually.Final Ventilation Rate and stir speed (S.S.) are respectively 5 liters/minute and 350 rev/mins.Cultivate after 72 hours, collect thalline.Dry cell weight reaches 8.12 grams per liters, cell content astaxanthin 783 micrograms/gram stem cell, and astaxanthin yield reaches 6358 micrograms per litre.
Embodiment 13
The amplification of embodiment 2 is to carry out in 5 liters of (New Brunswick Scientific, Bioflo3000 USA) bio-reactors, and liquid amount is 3.5 liters.Fermentation condition is with embodiment 3.When remaining sugar concentration in the nutrient solution is lower than 15 grams per liters, add 20 grams per liter molasses, along with the growth of cell, oxygen consumption rate increases gradually, in order to keep dissolved oxygen level 40%, increases Ventilation Rate and stir speed (S.S.) gradually.Final Ventilation Rate and stir speed (S.S.) are respectively 5 liters/minute and 500 rev/mins.Cultivate after 84 hours, collect thalline.Dry cell weight reaches 16.2 grams per liters, cell content astaxanthin 752 micrograms/gram stem cell, and astaxanthin yield reaches 12182 micrograms per litre.
Embodiment 14
Carry out in 5 liters of (New Brunswick Scientific, Bioflo3000 USA) bio-reactors, liquid amount is 3.5 liters.Other fermentation conditions are with embodiment 4.When remaining sugar concentration in the nutrient solution is lower than 15 grams per liters, add 60 grams per liter molasses, along with the growth of cell, oxygen consumption rate increases gradually, in order to keep dissolved oxygen level 40%, increases Ventilation Rate and stir speed (S.S.) gradually.Final Ventilation Rate and stir speed (S.S.) are respectively 5 liters/minute and 500 rev/mins.Cultivate after 96 hours, dry cell weight reaches 30.2 grams per liters, cell content astaxanthin 765 micrograms/gram stem cell, and astaxanthin yield reaches 23103 micrograms per litre; Cultivate after 120 hours, collect thalline, dry cell weight reaches 29.8 grams per liters, cell content astaxanthin 821 micrograms/gram stem cell, and astaxanthin yield reaches 24466 micrograms per litre.
Embodiment 15
Other conditions are added 10 grams per liter molasses with embodiment 11.Dry cell weight reaches 11.5 grams per liters, cell content astaxanthin 786 micrograms/gram stem cell, and astaxanthin yield reaches 9039 micrograms per litre.
Embodiment 16
Carry out in 5 liters of (New Brunswick Scientific, Bioflo3000 USA) bio-reactors, liquid amount is 3.5 liters.Other fermentation conditions are with embodiment 5.When remaining sugar concentration in the nutrient solution is lower than 15 grams per liters, add 60 grams per liter molasses, along with the growth of cell, oxygen consumption rate increases gradually, in order to keep dissolved oxygen level 40%, increases Ventilation Rate and stir speed (S.S.) gradually.Added 3 grams per liter dehydrated alcohols in 60 hours.Final Ventilation Rate and stir speed (S.S.) are respectively 5 liters/minute and 500 rev/mins.Cultivate after 120 hours, dry cell weight reaches 31.2 grams per liters, cell content astaxanthin 931 micrograms/gram stem cell, and astaxanthin yield reaches 29047 micrograms per litre.
Embodiment 17
Other conditions were added 8 grams per liter dehydrated alcohols in 16,60 hours with embodiment.The control dissolved oxygen level is 40%, and final Ventilation Rate and stir speed (S.S.) are respectively 5 liters/minute and 500 rev/mins.Cultivate after 120 hours, dry cell weight reaches 31.6 grams per liters, cell content astaxanthin 905 micrograms/gram stem cell, and astaxanthin yield reaches 28598 micrograms per litre.
Embodiment 18
Other conditions are controlled dissolved oxygen level 10% with embodiment 14.Dry cell weight reaches 22.6 grams per liters, cell content astaxanthin 714 micrograms/gram stem cell, and astaxanthin yield reaches 16136 micrograms per litre.
Embodiment 19
Other conditions are controlled dissolved oxygen level 80% with embodiment 14.Dry cell weight reaches 31.2 grams per liters, cell content astaxanthin 882 micrograms/gram stem cell, and astaxanthin yield reaches 27518 micrograms per litre.
Claims (5)
1, a kind of method of utilizing molasses or the middle fed-batch fermentation of Dian Fentang raw material to produce astaxanthin, it is characterized in that microbial inoculant in liquid nutrient medium, under 18-22 ℃ temperature, dissolved oxygen concentration is controlled at 10-80%, the pH of culturing process is 3-6, when remaining sugar concentration in the substratum is lower than 15 grams per liters, add molasses or Dian Fentang to substratum sugar concentration 10-60 grams per liter and continue to cultivate, after 72-120 hour, finish to cultivate; The microorganism that is used to inoculate is red Fife's yeast, and substratum is that molasses or Dian Fentang add inorganic or organic nitrogen source.
2, according to a kind of method of utilizing molasses or the middle fed-batch fermentation of Dian Fentang raw material to produce astaxanthin described in the claim 1, it is characterized in that described red Fife's yeast is As2.1557.
3, a kind of method that fed-batch fermentation is produced astaxanthin in the middle of molasses or the Dian Fentang raw material of utilizing according to claim 2, it is characterized in that described inorganic nitrogen-sourced be ammonium sulfate, ammonium nitrate, ammonium chloride or urea.
4, a kind of method that fed-batch fermentation is produced astaxanthin in the middle of molasses or the Dian Fentang raw material of utilizing according to claim 3 is characterized in that described organic nitrogen source is yeast powder or yeast extract paste, peptone, corn steep liquor.
5, utilize the method that fed-batch fermentation is produced astaxanthin in the middle of molasses or the Dian Fentang raw material according to a kind of described in the claim 4, it is characterized in that described culturing process added ethanol, consumption is 0-8g/L.
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| CN 200310112328 CN1282749C (en) | 2003-11-25 | 2003-11-25 | Course feeding fermentation method for preparing astaxanthin with molasses or starch sugar as materials |
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| CN 200310112328 CN1282749C (en) | 2003-11-25 | 2003-11-25 | Course feeding fermentation method for preparing astaxanthin with molasses or starch sugar as materials |
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| CN1282749C true CN1282749C (en) | 2006-11-01 |
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Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100386426C (en) * | 2006-01-05 | 2008-05-07 | 大连轻工业学院 | Method for cultivating phaffiafhodozyma enriched with astaxanthin using waste water from bean curd production |
| CN101579040B (en) * | 2009-06-14 | 2011-10-12 | 中国热带农业科学院热带作物品种资源研究所 | Preparation method of astaxanthin biological feed |
| CN101703152B (en) * | 2009-10-30 | 2012-12-26 | 华南理工大学 | Method for preparing astaxanthin feed additive by using beer spent grains |
| CN103820520B (en) * | 2014-03-06 | 2017-02-01 | 浙江皇冠科技有限公司 | High-yield natural astaxanthin fermentation method |
| CN105695550B (en) * | 2016-03-31 | 2019-12-17 | 广州元大生物科技发展有限公司 | Method for producing astaxanthin by high-density culture of lactobacillus plantarum |
| CN107164449B (en) * | 2017-08-03 | 2021-06-25 | 广州立达尔生物科技股份有限公司 | Method for producing astaxanthin by fermenting marigold flower meal extracting solution as raw material and application |
| CN111155975B (en) * | 2020-01-19 | 2022-05-03 | 山西大学 | Method for improving yield of biological coal bed gas through feed supplement fermentation |
| CN114085881B (en) * | 2020-08-25 | 2023-12-22 | 浙江医药股份有限公司新昌制药厂 | Method for improving yield of Phaffia rhodozyma astaxanthin and application thereof |
| CN112063539A (en) * | 2020-09-16 | 2020-12-11 | 厦门昶青生物科技有限公司 | Culture medium and culture method for producing carotenoid by fermentation of phaffia rhodozyma |
| CN114045226A (en) * | 2021-11-19 | 2022-02-15 | 山东省食品发酵工业研究设计院 | Low-cost culture medium for culturing phaffia rhodozyma producing astaxanthin and preparation method and application thereof |
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