CN104830938A - Method for enhancing mussel astaxanthin fermentation production yield - Google Patents
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Abstract
The invention discloses a method for enhancing mussel astaxanthin fermentation production yield, which comprises the following steps: by using rhodotorula mucilaginosa as a fermentation strain, inoculating 2-10% of the rhodotorula mucilaginosa into a culture medium, and adding calcium carbonate and magnesium sulfate into the culture medium, wherein the addition amount of the calcium carbonate is 0.1-0.5 g/L, and the magnesium sulfate is magnesium sulfate heptahydrate with the addition amount of 0.1-0.7 g/L; and fermenting in a 5L fermentation tank for 3 days while regulating the ventilation quantity and rotation speed of the fermentation tank and keeping the fermentation temperature at 25-33 DEG C and the pH value at 5.5-7.5. On the premise of ensuring the product yield, the method can lower the energy consumption and reduce the fermentation operating cost by controlling the ventilation quantity and rotation speed in different fermentation stages, and thus, has important meanings for reducing industrialized production cost.
Description
Technical field
The present invention relates to microbial technology field, specifically, relate to a kind of method improving the red astaxanthin productive rate in fermentative production sea.
Background technology
The red astaxanthin in sea is a Carotenoids, has extremely strong resistance of oxidation and biological tinting strength, has been widely used in many fields such as food, medicine, makeup, feed.In prevention cardiovascular and cerebrovascular diseases, protection eyes and central nervous system, ultraviolet radiation preventing, Tumor suppression, anti-inflammatory, relieving sports fatigue etc., there is higher using value.The red astaxanthin in sea, a kind of carotenoid with high anti-oxidation ability, during thin-layer chromatography its mobility and natural astaxanthin ester close, resistance of oxidation also with natural left-handed astaxanthin close to and gain the name, DPPH method detects its resistance of oxidation and is better than natural left-handed astaxanthin about 40%.
The preparation method of astaxanthin, there will be a known chemical synthesis, the method extracted from the extraction of aquatic products refuse or algae, utilizes microorganism to carry out the method etc. of producing.Chemical synthesis, production cost is higher, and many countries do not allow to use synthesizing astaxanthin for fodder additives aspect; Extract from aquatic products refuse, raw material sources are extensive, cost is low, but production is easily subject to seasonal restrictions, and working condition requires harsh, and production cost is high, and output is lower, and product purity is not high; Extract from algae, the content astaxanthin in algae is higher, and raw materials cost is lower.But the growth conditions of algae is harsh, very high to the requirement of water quality, environment and light, still there is difficulty in scale operation; Utilize microorganism to carry out the method for producing, adopt saccharine material fermentative production, be beneficial to large scale and high density and produce, and be not subject to the restriction of region and weather, production cost relative moderate.
In recent years, increasing to the demand of astaxanthin both at home and abroad, there are wide market outlook.Many biotech companies of the states such as the current U.S., Canada, European Union are just being devoted to this series products of Development and Production, but far can not meet the need of market, therefore price is higher.Therefore stepping up the development to astaxanthin and analogous products, for enhancing our international competitiveness, being of great significance.
Summary of the invention
For solving the problems of the technologies described above, the invention provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be that described rhodotorula mucilaginosa is seeded in substratum by 2 – 10% according to inoculum size, described substratum comprises: glucose 60-130g/L, yeast powder 3-10g/L, KH
2pO
40.5-1.5g/L, K
2hPO
40.5-1.5g/L, urea 1-5g/L and NaCl 0.3-9g/L, the pH of this substratum is between 5.5-7.5, calcium carbonate and magnesium sulfate is added in described substratum, the addition of described calcium carbonate is 0.1-0.5g/L, and described magnesium sulfate is bitter salt, and addition is 0.1-0.7g/L, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 25-33 DEG C, and fermentation pH value is at the indirect fermentation 3d of 5.5-7.5;
Described ventilation, is further,
In 1-12h during the fermentation, ventilation is 1.5-1.75L/min;
In 12-36h during the fermentation, ventilation is 6.5-9L/min;
In 36-72h during the fermentation, ventilation is 2-3.5L/min;
Described rotating speed, is further,
In 1-12h during the fermentation, rotating speed is 140-180r/min;
In 12-36h during the fermentation, rotating speed is 220-280r/min;
In 36-72h during the fermentation, rotating speed is 80-180r/min.
Preferably, described substratum comprises further: glucose 70g/L, yeast powder 5g/L, KH
2pO
41g/L, K
2hPO
41g/L, urea 2g/L and NaCl 2g/L, the pH of this substratum is 6.5.
Rhodotorula mucilaginose of the present invention (English name Rhodotorula mucilaginosa), be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, preservation date: on December 01st, 2014, deposit number: CGMCC No.10123.
Compared with prior art, the method for the red astaxanthin productive rate in raising fermentative production sea of the present invention, reaches following effect:
(1) the present invention take rhodotorula mucilaginosa as starting strain, and glucose etc. are main production raw material, adopts 5L ferment tank, changes fermentative medium formula on the original basis, regulates fermentation condition to make the content of extra large red astaxanthin in thalline add 10.34%.
(2) take rhodotorula mucilaginosa as starting strain, glucose and yeast powder are main production raw material, by adding a certain amount of calcium carbonate and magnesium sulfate, and regulate ventilation and the rotating speed of different steps during the fermentation, in dry mycelium, extra large red content astaxanthin reaches as high as 1.28% ~ 3.75%.
(3) the present invention is on the basis ensureing product yield, by ventilation and rotating speed in the control in different fermentations stage, can energy consumption be reduced, reduce fermentation operation cost, larger meaning is existed for the reduction realizing industrial production cost.
Accompanying drawing explanation
Accompanying drawing described herein is used to provide a further understanding of the present invention, forms a part of the present invention, and schematic description and description of the present invention, for explaining the present invention, does not form inappropriate limitation of the present invention.In the accompanying drawings:
Fig. 1 is the impact of calcium carbonate addition on the red astaxanthin yield in sea;
Fig. 2 is the impact of bitter salt on the red astaxanthin yield in sea;
Fig. 3 is the method flow diagram that the present invention improves the red astaxanthin productive rate in fermentative production sea.
Embodiment
As employed some vocabulary to censure specific components in the middle of specification sheets and claim.Those skilled in the art should understand, and hardware manufacturer may call same assembly with different noun.This specification and claims are not used as with the difference of title the mode distinguishing assembly, but are used as the criterion of differentiation with assembly difference functionally." comprising " as mentioned in the middle of specification sheets and claim is in the whole text an open language, therefore should be construed to " comprise but be not limited to "." roughly " refer to that in receivable limit of error, those skilled in the art can solve the technical problem within the scope of certain error, reach described technique effect substantially.In addition, " couple " word and comprise directly any and indirectly electric property coupling means at this.Therefore, if describe a first device in literary composition to be coupled to one second device, then represent described first device and directly can be electrically coupled to described second device, or be indirectly electrically coupled to described second device by other devices or the means that couple.Specification sheets subsequent descriptions is for implementing better embodiment of the present invention, and right described description is to illustrate for the purpose of rule of the present invention, and is not used to limit scope of the present invention.Protection scope of the present invention is when being as the criterion depending on the claims person of defining.
Below in conjunction with accompanying drawing, the present invention is described in further detail, but not as a limitation of the invention.
Embodiment 1:
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 2% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 60g/L, yeast powder 3g/L, KH
2pO
40.5g/L, K
2hPO
40.5g/L, urea 1g/L and NaCl0.3g/L, the pH of this substratum is 6.5, in described substratum, add calcium carbonate and magnesium sulfate, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 25 DEG C, and fermentation pH value 5.5 is fermented 3d.
The addition of described calcium carbonate is 0.1g/L, and described magnesium sulfate is bitter salt, and addition is 0.1-0.7g/L.
Described ventilation, is further, and in 1-12h during the fermentation, ventilation is 1.5L/min; In 12-36h during the fermentation, ventilation is 7L/min; In 36-72h during the fermentation, ventilation is 2L/min.
Described rotating speed, is further, and in 1-12h during the fermentation, rotating speed is 140r/min; In 12-36h during the fermentation, rotating speed is 220r/min; In 36-72h during the fermentation, rotating speed is 140r/min.
Embodiment 2:
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 10% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 130g/L, yeast powder 10g/L, KH
2pO
41.5g/L, K
2hPO
41.5g/L, urea 5g/L and NaCl9g/L, the pH of this substratum is 7.5, in described substratum, add calcium carbonate and magnesium sulfate, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 25 DEG C, and pH value 7.5 is fermented 3d.
The addition of described calcium carbonate is 0.5g/L, and described magnesium sulfate is bitter salt, and addition is 0.7g/L.
Described ventilation, is further, and in 1-12h during the fermentation, ventilation is 1.75L/min; In 12-36h during the fermentation, ventilation is 9L/min; In 36-72h during the fermentation, ventilation is 3.5L/min.
Described rotating speed, is further, and in 1-12h during the fermentation, rotating speed is 180r/min; In 12-36h during the fermentation, rotating speed is 280r/min; In 36-72h during the fermentation, rotating speed is 180r/min.
Embodiment 3:
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 6% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 95g/L, yeast powder 7g/L, KH
2pO
41g/L, K
2hPO
41g/L, urea 3g/L and NaCl4.65g/L, the pH6.5 of this substratum, adds calcium carbonate and magnesium sulfate in described substratum, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 31 DEG C, and fermentation pH value 6.5 is fermented 3d.
The addition of described calcium carbonate is 0.3g/L, and described magnesium sulfate is bitter salt, and addition is 0.4g/L.
Described ventilation, in 1-12h during the fermentation, ventilation is 1.65L/min; In 12-36h during the fermentation, ventilation is 8L/min; In 36-72h during the fermentation, ventilation is 2.8L/min.
Described rotating speed, in 1-12h during the fermentation, rotating speed is 160r/min; In 12-36h during the fermentation, rotating speed is 250r/min; In 36-72h during the fermentation, rotating speed is 160r/min.
Embodiment 4:
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 3% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 70g/L, yeast powder 5g/L, KH
2pO
41g/L, K
2hPO
41g/L, urea 2g/L and NaCl2g/L, the pH6.5 of this substratum, adds calcium carbonate and magnesium sulfate in described substratum, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 30 DEG C, and fermentation pH value 6.5 is fermented 3d.
The addition of described calcium carbonate is 0.1g/L, and described magnesium sulfate is bitter salt, and addition is 0.1g/L.
Described ventilation, in 1-12h during the fermentation, ventilation is 1.75L/min; In 12-36h during the fermentation, ventilation is 7L/min; In 36-72h during the fermentation, ventilation is 3.5L/min.
Described rotating speed, in 1-12h during the fermentation, rotating speed is 180r/min; In 12-36h during the fermentation, rotating speed is 280r/min; In 36-72h during the fermentation, rotating speed is 180r/min.
Embodiment 5:
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 4% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 85g/L, yeast powder 5g/L, KH
2pO
41.8g/L, K
2hPO
41.8g/L, urea 2.4g/L and NaCl2.8g/L, the pH6 of this substratum, adds calcium carbonate and magnesium sulfate in described substratum, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 30 DEG C, and fermentation pH value 6 is fermented 3d.
The addition of described calcium carbonate is 0.3g/L, and described magnesium sulfate is bitter salt, and addition is 0.3g/L.
Described ventilation, in 1-12h during the fermentation, ventilation is 1.63L/min; In 12-36h during the fermentation, ventilation is 8.4L/min; In 36-72h during the fermentation, ventilation is 2.8L/min.
Described rotating speed, in 1-12h during the fermentation, rotating speed is 180r/min; In 12-36h during the fermentation, rotating speed is 280r/min; In 36-72h during the fermentation, rotating speed is 180r/min.
Embodiment 6:
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 4% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 86g/L, yeast powder 7g/L, KH
2pO
40.6g/L, K
2hPO
40.6g/L, urea 2g/L and NaCl2g/L, the pH6.3 of this substratum, adds calcium carbonate and magnesium sulfate in described substratum, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 26 DEG C, and fermentation pH value 6.3 is fermented 3d.
The addition of described calcium carbonate is 0.4g/L, and described magnesium sulfate is bitter salt, and addition is 0.4g/L.
Described ventilation, in 1-12h during the fermentation, ventilation is 1.72L/min; In 12-36h during the fermentation, ventilation is 8.2L/min; In 36-72h during the fermentation, ventilation is 3.1L/min.
Described rotating speed, in 1-12h during the fermentation, rotating speed is 175r/min; In 12-36h during the fermentation, rotating speed is 265r/min; In 36-72h during the fermentation, rotating speed is 175r/min.
Embodiment 7
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 4.6% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 77g/L, yeast powder 7g/L, KH
2pO
40.8g/L, K
2hPO
40.8g/L, urea 3g/L and NaCl3g/L, the pH6.2 of this substratum, adds calcium carbonate and magnesium sulfate in described substratum, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 28 DEG C, and fermentation pH value 6.2 is fermented 3d.
The addition of described calcium carbonate is 0.5g/L, and described magnesium sulfate is bitter salt, and addition is 0.5g/L.
Described ventilation, in 1-12h during the fermentation, ventilation is 1.64L/min; In 12-36h during the fermentation, ventilation is 7.93L/min; In 36-72h during the fermentation, ventilation is 2.86L/min.
Described rotating speed, in 1-12h during the fermentation, rotating speed is 175r/min; In 12-36h during the fermentation, rotating speed is 265r/min; In 36-72h during the fermentation, rotating speed is 175r/min.
Embodiment 8
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 4.8% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 69g/L, yeast powder 3g/L, KH
2pO
40.85g/L, K
2hPO
40.85g/L, urea 2g/L and NaCl2g/L, the pH5.5 of this substratum, adds calcium carbonate and magnesium sulfate in described substratum, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 26 DEG C, and fermentation pH value 5.5 is fermented 3d.
The addition of described calcium carbonate is 0.3g/L, and described magnesium sulfate is bitter salt, and addition is 0.7g/L.
Described ventilation, in 1-12h during the fermentation, ventilation is 1.68L/min; In 12-36h during the fermentation, ventilation is 7-9L/min; In 36-72h during the fermentation, ventilation is 2-3.5L/min.
Described rotating speed, in 1-12h during the fermentation, rotating speed is 160r/min; In 12-36h during the fermentation, rotating speed is 250r/min; In 36-72h during the fermentation, rotating speed is 160r/min.
Embodiment 9
The present embodiment provides a kind of method improving the red astaxanthin productive rate in fermentative production sea, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be 3.7% be seeded in substratum by described rhodotorula mucilaginosa according to inoculum size, described substratum comprises: glucose 72g/L, yeast powder 5g/L, KH
2pO
40.7g/L, K
2hPO
40.7g/L, urea 2g/L and NaCl2g/L, the pH5.8 of this substratum, adds calcium carbonate and magnesium sulfate in described substratum, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 27 DEG C, and fermentation pH value 5.8 is fermented 3d.
The addition of described calcium carbonate is 0.2g/L, and described magnesium sulfate is bitter salt, and addition is 0.6g/L.
Described ventilation, in 1-12h during the fermentation, ventilation is 1.7L/min; In 12-36h during the fermentation, ventilation is 8L/min; In 36-72h during the fermentation, ventilation is 3.0L/min.
Described rotating speed, in 1-12h during the fermentation, rotating speed is 150r/min; In 12-36h during the fermentation, rotating speed is 240r/min; In 36-72h during the fermentation, rotating speed is 150r/min.
Embodiment 10
Composition graphs 1, the implementation case illustrates that calcium carbonate concentration produces the impact of extra large red astaxanthin yield to rhodotorula mucilaginosa, and on the basis of embodiment 4, change the addition of calcium carbonate, other experiment condition is identical.On the basis of fermention medium, adding concentration is respectively the calcium carbonate of 0g/L, 0.1g/L, 0.2g/L, 0.3g/L, 0.4g/L or 0.5g/L, and leavening temperature is 30 DEG C, and rotating speed is 280r/min, fermentation 3d.
As shown in Figure 1, economically analyze, when interpolation calcium carbonate concentration is 0.2g/L, the content of extra large red astaxanthin reaches the highest.
Embodiment 11
Composition graphs 2, on the basis of embodiment 4, change the addition of bitter salt, other experiment condition is identical.The implementation case illustrates that magnesium sulfate concentration produces the impact of extra large red astaxanthin yield to rhodotorula mucilaginosa, on the basis of fermention medium, adding concentration is respectively the bitter salt of 0g/L, 0.1g/L, 0.2g/L, 0.3g/L, 0.4g/L, 0.5g/L, 0.6g/L or 0.7g/L, leavening temperature is 30 DEG C, rotating speed is 280r/min, fermentation 3d.
As shown in Figure 2, economically analyze, when interpolation bitter salt concentration is 0.5g/L, the content of extra large red astaxanthin reaches the highest.
Embodiment 12
On the basis of embodiment 4, the implementation case illustrates on the basis ensureing thalline productive rate and the red astaxanthin productive rate in sea, and by regulating ventilation and rotating speed in fermenting process to control dissolved oxygen, other experiment condition is identical.Rhodotorula mucilaginosa is aerobic thalline, and at the 1-12h of fermentation, now cell concentration is lower, relatively less to the demand of oxygen, and therefore, now controlling ventilation is 1.75L/min, and rotating speed 180r/min, just can ensure the demand of thalline to oxygen; When fermenting 12-36h, thalline enters logarithmic phase, and metabolism is very vigorous, during this oxygen-consumption along with fermentation carrying out increase fast, now ventilation is adjusted to 7L/min, just can meet the demand to oxygen of the thalline when arriving oxygen consumption vertex when rotating speed is 280r/min; Enter stationary phase to thalline during fermentation 36-72h, metabolism is slowed down, and oxygen-consumption starts to reduce, and dissolved oxygen amount starts to rise, and therefore, turns down ventilation and rotating speed, be respectively 3.5L/min and 180r/min after 36h, can ensure that this stage oxygen dissolving maintains 40% ~ 65%.On the basis ensureing product yield, by ventilation and rotating speed in the control in different fermentations stage, can energy consumption be reduced, reduce fermentation operation cost, larger meaning is existed for the reduction realizing industrial production cost.
Above-mentioned explanation illustrate and describes some preferred embodiments of the present invention, but as previously mentioned, be to be understood that the present invention is not limited to the form disclosed by this paper, should not regard the eliminating to other embodiments as, and can be used for other combinations various, amendment and environment, and can in invention contemplated scope described herein, changed by the technology of above-mentioned instruction or association area or knowledge.And the change that those skilled in the art carry out and change do not depart from the spirit and scope of the present invention, then all should in the protection domain of claims of the present invention.
Claims (2)
1. one kind is improved the method for the red astaxanthin productive rate in fermentative production sea, it is characterized in that, realized by following steps: using Rhodotorula mucilaginose as fermented bacterium, be that described rhodotorula mucilaginosa is seeded in substratum by 2 – 10% according to inoculum size, described substratum comprises: glucose 60-130g/L, yeast powder 3-10g/L, KH
2pO
40.5-1.5g/L, K
2hPO
40.5-1.5g/L, urea 1-5g/L and NaCl 0.3-9g/L, the pH of this substratum is between 5.5-7.5, calcium carbonate and magnesium sulfate is added in described substratum, the addition of described calcium carbonate is 0.1-0.5g/L, and described magnesium sulfate is bitter salt, and addition is 0.1-0.7g/L, the fermentor tank of 5L is adopted to ferment, regulate ventilation and fermentor tank rotating speed during the fermentation, leavening temperature maintains 25-33 DEG C, and fermentation pH value is at the indirect fermentation 3d of 5.5-7.5;
Described ventilation is, in 1-12h during the fermentation, ventilation is 1.5-1.75L/min; In 12-36h during the fermentation, ventilation is 6.5-9L/min; In 36-72h during the fermentation, ventilation is 2-3.5L/min;
Described rotating speed is, in 1-12h during the fermentation, rotating speed is 140-180r/min; In 12-36h during the fermentation, rotating speed is 220-280r/min; In 36-72h during the fermentation, rotating speed is 80-180r/min.
2. the method for the red astaxanthin productive rate in raising fermentative production sea according to claim 1, it is characterized in that, described substratum comprises further: glucose 70g/L, yeast powder 5g/L, KH
2pO
41g/L, K
2hPO
41g/L, urea 2g/L and NaCl 2g/L, the pH of this substratum is 6.5.
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| CN108998493A (en) * | 2018-06-25 | 2018-12-14 | 浙江皇冠科技有限公司 | A kind of formula technique of high-yield astaxanthin fermentation medium and application |
| CN108998493B (en) * | 2018-06-25 | 2020-11-06 | 浙江皇冠科技有限公司 | Formula technology and application of fermentation medium for high-yield astaxanthin |
| CN110964676A (en) * | 2020-01-04 | 2020-04-07 | 山东得和明兴生物科技有限公司 | Method for culturing high-bacterial-quantity fermentation liquor of brevibacillus laterosporus |
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