CN109662224B - Fermentation production method of apple juice rich in coenzyme Q10 and coenzyme Q10 apple juice - Google Patents
Fermentation production method of apple juice rich in coenzyme Q10 and coenzyme Q10 apple juice Download PDFInfo
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- CN109662224B CN109662224B CN201910018091.4A CN201910018091A CN109662224B CN 109662224 B CN109662224 B CN 109662224B CN 201910018091 A CN201910018091 A CN 201910018091A CN 109662224 B CN109662224 B CN 109662224B
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- 235000017471 coenzyme Q10 Nutrition 0.000 title claims abstract description 72
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
- A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention discloses a fermentation production method of apple juice rich in coenzyme Q10 and coenzyme Q10 apple juice. The preparation method comprises the following steps: fresh squeezed apple juice is used as raw material, 10% rhodobacter sphaeroides is added, and fermentation is carried out for 72h under the conditions that the fermentation temperature is 35 ℃ and the rotating speed of a shaking table is 180r/min, so as to prepare the fermented apple juice rich in coenzyme Q10, wherein the amount of rhodobacter sphaeroides reaches 8.7 multiplied by 107CFU/mL, the content of coenzyme Q10 reaches 9.54 mg/L. The fruit juice prepared by the fermentation process disclosed by the invention is light red in color, sweet in taste, soft in taste, rich in coenzyme Q10 and viable count, and is a novel fermented apple juice product with a certain antioxidant function.
Description
Technical Field
The invention belongs to the technical field of microbial fermentation, and particularly relates to a fermentation production method of apple juice rich in coenzyme Q10 and coenzyme Q10 apple juice.
Background
The health care function and the extremely advantageous flavor of the probiotic fermented juice are increasingly valued by people's pursuit and researchers. At present, strains adopted by probiotic fermentation of fruit juice comprise strains of lactobacillus acidophilus, bifidobacterium, lactobacillus plantarum and the like, and the probiotic fermentation can improve the utilization rate of metal elements (calcium, iron and phosphorus) and promote iron and VDThe soft sour taste brought by the absorption of the fermented fruit juice can improve the flavor of the fermented fruit juice, and simultaneously has the effects of regulating the micro-ecological balance in the intestinal tract of a human body, regulating the function of an immune system and delaying the aging of the body. In addition, researchers are also searching for new probiotic strains, developing fermented apple juice products with larger health care functions and solving the problem of processing diversification of apple juice.
The rhodobacter sphaeroides has the efficacy of resisting the toxic reaction of organisms caused by bacterial endotoxin and can improve the immune function of the organisms. In addition, coenzyme Q10 produced by rhodobacter sphaeroides can activate body, increase ATP production, treat various human diseases, such as cardiomyopathy, muscular dystrophy, periodontal disease, diabetes, atherosclerosis, etc., delay aging, increase nonspecific immunity of human body, and maintain the integrity of biological membrane.
Disclosure of Invention
Aiming at the species of the strains of the existing fermented fruit juice, the invention provides a fermentation production method of apple juice rich in coenzyme Q10 and the coenzyme Q10 apple juice, and the apple juice with higher coenzyme Q10 content can be obtained by the fermentation production method.
The technical scheme adopted by the invention is as follows:
a fermentation production method of apple juice rich in coenzyme Q10 comprises the steps of taking the apple juice as a fermentation raw material, and taking rhodobacter sphaeroides as a fermentation strain; the inoculation amount of rhodobacter sphaeroides seed bacterial liquid is 10 percent according to the volume of the apple juice.
Optionally, activating rhodobacter sphaeroides to obtain rhodobacter sphaeroides activation 1 generation culture solution, and inoculating 10% of rhodobacter sphaeroides activation 1 generation culture solution into the apple juice according to the volume of the apple juice to obtain rhodobacter sphaeroides seed bacterial solution.
Optionally, the preparation of rhodobacter sphaeroides seed bacterial liquid comprises:
a. and (3) strain activation culture: taking 200uL of rhodobacter sphaeroides from a glycerol tube, and standing and culturing the rhodobacter sphaeroides in an activation culture medium at the temperature of 30 ℃ for 48 hours to prepare an activation 1-generation culture solution of rhodobacter sphaeroides;
b. seed liquid culture: according to the volume of the apple juice, 10 percent of activated 1-generation culture solution is added into an apple juice culture medium, and dark culture is carried out for 48 hours in a shaking table at 30 ℃ and 150r/min to obtain rhodobacter sphaeroides seed bacterial solution.
Optionally, the activation medium: according to the addition amount of each liter of water, 20g/L of glucose, 10g/L of yeast extract, 10g/L of peptone, 0.05g/L of magnesium sulfate, 0.05g/L of monopotassium phosphate and 0.05g/L of dipotassium phosphate.
Optionally, the fermentation temperature of the fermentation is 30-35 ℃, and the rotating speed of the shaking table is 150-200 r/min.
Optionally, the fermentation temperature of the fermentation is 35 ℃, and the rotating speed of the shaking table is 180 r/min.
Optionally, the fermentation temperature of the fermentation is 35 ℃, the rotating speed of the shaking table is 180r/min, and the fermentation time is 72 h.
Optionally, the preparation of the apple juice comprises: juicing the apple blocks, adding 450mg/L pectinase for enzymolysis at 45 ℃ for 2h, and filtering to obtain apple juice.
The coenzyme Q10 apple juice is prepared by the method of the invention, and the coenzyme Q10 apple juice is obtained by the method of the invention.
Compared with the prior art, the invention has the advantages that:
compared with the traditional fermentation strains, such as lactic acid bacteria, acetic acid bacteria, saccharomycetes, bifidobacteria and the like, the rhodobacter sphaeroides is rich in the nutritive values of coenzyme Q10, protein, carotenoid and the like, and the original effect of a converted substance can be amplified to a certain degree due to the complex biotransformation function of the rhodobacter sphaeroides. The apple juice is fermented by rhodobacter sphaeroides, so that the nutritive value of rhodobacter sphaeroides and coenzyme Q10 can be increased for the apple juice, and the health-care functions of the apple juice such as oxidation resistance and the like can be improved.
Drawings
FIG. 1 comparison of the Dry weight of the cells with the content of coenzyme Q10 (S for Rhodospirillum rubrum, Z for Rhodopseudomonas palustris, L for rhodobacter sphaeroides)
FIG. 2 comparison of antioxidant capacity (S for Rhodospirillum rubrum, Z for Rhodopseudomonas palustris, L for rhodobacter sphaeroides, G for apple juice)
FIG. 3 effect of inoculum size on viable count, coenzyme Q10 and sensory score;
FIG. 4 effect of liquid loading on viable count, coenzyme Q10 and sensory score;
FIG. 5 effect of temperature on viable count, coenzyme Q10 and sensory score;
FIG. 6 the effect of the shaker speed on viable count, coenzyme Q10 and sensory score;
FIG. 7 dynamic changes in pH, total acid in fermented apple juice;
FIG. 8 dynamic changes in SSC, total sugar in fermented apple juice;
FIG. 9 dynamic change of total phenolic content in fermented apple juice;
FIG. 10 dynamic change in antioxidant capacity of fermented apple juice;
the invention is described in detail below with reference to the drawings and the detailed description.
Detailed Description
The method takes fresh apple juice as a raw material, adopts a single-factor test and an orthogonal optimization test to optimize the process of fermenting the apple juice by rhodobacter sphaeroides, takes the viable count, the coenzyme Q10 content and the sensory evaluation of the fermented apple juice as evaluation indexes, then measures the physicochemical index of the fermented apple juice under the optimized condition, analyzes the dynamic change of the physicochemical index of the fermented apple juice, and aims to regulate and control the sensory quality of the fermented apple juice.
Rhodobacter sphaeroides is rich in proteins, B vitamins, coenzyme Q10 and complex biotransformation functions. Coenzyme Q10 produced by rhodobacter sphaeroides is a fat-soluble quinone substance, has the functions of activating the body, increasing the production of ATP, controlling the oxygen flow in cells, delaying senescence, resisting oxidation, increasing the non-specific immunity of a human body, maintaining the integrity of a biological membrane, and is helpful for treating various human diseases such as cardiomyopathy, muscular dystrophy, periodontal disease, diabetes, atherosclerosis and the like. Rhodobacter sphaeroides fermented apple juice can utilize the biotransformation function of rhodobacter sphaeroides and the components of rhodobacter sphaeroides per se to increase or enhance the nutritive value and health care function of the apple juice.
The procedure at the time of the test was as follows:
(1) strain screening
Adding Rhodopseudomonas palustris 1.2349(Rhodopseudomonas palustris), rhodobacter sphaeroides 1.2182(rhodobacter sphaeroides) and rhodospirillum rubrum 1.5005 (rhodospirillum) 3 photosynthetic bacteria into apple juice for fermentation culture, taking the apple juice fermented at constant temperature for 72 hours to measure indexes such as biomass, coenzyme Q10 content, oxidation resistance and the like, taking the apple juice without added strains as a blank control, and screening suitable strains of the fermented apple juice.
As can be seen from FIG. 1 (in FIG. 1, letters a, B, C, A, B, C, etc. indicate significance of one-way analysis of variance comparison), Coq indicates coenzyme Q10, DCW indicates biomass, biomass and coenzyme Q10 content of rhodobacter sphaeroides are highest, and rhodopseudomonas palustris and rhodospirillum rubrum are sequentially. The remarkable difference (P is less than 0.5) exists among 3 strains, which indicates that rhodobacter sphaeroides in apple juice has stronger vitality than rhodopseudomonas palustris and rhodospirillum rubrum and the coenzyme Q10 yield is high. As can be seen from FIG. 2 (the letters a, b, c, etc. in FIG. 2 indicate the significance of the one-way ANOVA comparative differences), rhodobacter sphaeroides fermented apple juice has significantly higher antioxidant capacity than apple juice, while Rhodopseudomonas palustris and Rhodospirillum rubrum are significantly lower than apple juice. The rhodobacter sphaeroides is found by comparison to be a photosynthetic bacterium which is suitable for fermenting apple juice, produces the largest amount of coenzyme Q10 and has the strongest juice oxidation resistance.
(2) Process flow
Fresh apple juice → rhodobacter sphaeroides seed bacteria liquid → fermentation under different conditions → determination of viable count, coenzyme Q10 content, sensory score → determination of physicochemical indexes under optimized conditions for dynamic analysis.
(3) Preparation of apple juice
Fully cleaning, cutting into pieces and removing cores of fully mature apples without rotting and deterioration, respectively juicing after soaking in a color fixative (0.05% Vc) for 15min, and then adding a certain amount of pectinase (450mg/L) for enzymolysis, wherein the enzymolysis conditions are as follows: carrying out enzymolysis at 45 ℃ for 2h, and carrying out suction filtration to obtain clear apple juice. Sterilizing at 105 deg.C for 15 min.
(4) Preparation of fermented apple juice
Preparing rhodobacter sphaeroides seed bacterial liquid: inoculating rhodobacter sphaeroides strains into a test tube filled with 10mL of activation culture medium, standing and culturing for 24h at 30 ℃, then transferring into a 250mL triangular flask filled with 100mL of apple juice culture medium, and performing secondary activation culture for 24h at 30 ℃ and 150r/min to obtain rhodobacter sphaeroides seed bacterial liquid.
Preparing fermented apple juice: inoculating 10% rhodobacter sphaeroides seed bacterial liquid into apple juice for fermentation. The apple juice is sterilized at 105 deg.C for 15 min.
(5) Fermentation Process Single factor test
The fermentation capacity of rhodobacter sphaeroides fermented apple juice is reflected by the viable count, the coenzyme Q10 yield and sensory score, and the influence of different inoculation amounts, liquid loading amounts, fermentation temperatures and table rotation speeds on the viable count, the coenzyme Q10 yield and the sensory score of rhodobacter sphaeroides is respectively inspected
Influence of inoculation amount on viable count, coenzyme Q10 and sensory score
Weighing 100mL of apple juice in a 250mL shake flask, sterilizing at 105 ℃ for 15min, respectively inoculating 8mL, 9mL, 10mL, 11mL and 12mL of rhodobacter sphaeroides seed bacterial liquid in the shake flask, and fermenting in a double-layer shaking table with the fermentation temperature of 30 ℃ and the rotation speed of the shaking table of 150 r/min. The effect of different inoculum amounts on viable count, coenzyme Q10 and sensory score were compared. The results are shown in FIG. 3.
As can be seen from FIG. 3, when the inoculation amount is increased from 8mL to 10mL, the viable count appears to be obviously increased, when the inoculation amount is 10mL, the viable count of the apple juice and the content of coenzyme Q10 are highest, and the viable count reaches 1.46 multiplied by 107CFU/mL, coenzyme Q10 content of 2.92mg/L, which is significantly higher than other groups (P)<0.05), when the inoculation amount is more than 10mL, the viable count and the coenzyme Q10 content are both reduced, the sensory score is gradually reduced along with the increase of the inoculation amount, and the differences of the three indexes are obvious through single-factor variance analysis (P)<0.05), namely the inoculation amount has great influence on the viable count, the sensory quality and the coenzyme Q10 content. The inoculum size of 9mL, 10mL, 11mL was selected for orthogonal optimization.
② the influence of liquid loading amount on viable count, coenzyme Q10 yield and sensory score
Respectively weighing 80, 90, 100, 110 and 120mL of apple juice, placing in a 250mL shake flask, and sterilizing at 105 deg.C for 15 min. Inoculating 10mL rhodobacter sphaeroides seed bacteria liquid into a shake flask, fermenting in a double-layer shaking table at the fermentation temperature of 30 ℃ and the rotation speed of 150r/min, and comparing the influences of different liquid contents on the viable count, the output of coenzyme Q10 and sensory score. The results are shown in FIG. 4.
As can be seen from fig. 4, when the liquid loading amount is 100mL, the number of living photosynthetic bacteria and the content of coenzyme Q10 are significantly higher than other liquid loading amounts (p <0.05), the sensory score is highest when the liquid loading amount is 120mL, the three indexes all have significant differences (p <0.05) through one-factor variance analysis, the advantage levels of the three indexes are comprehensively considered, and finally, 90mL, 100mL and 110mL of liquid loading amounts are selected for orthogonal optimization.
Influence of temperature on viable count, coenzyme Q10 yield and sensory score
100mL of apple juice is measured and placed in a 250mL shake flask, sterilization is carried out for 15min at 105 ℃, 10mL of rhodobacter sphaeroides target bacterial liquid is inoculated, fermentation culture is carried out in a shaking table with the rotating speed of 150r/min and the temperature of 25 ℃, 28 ℃, 30 ℃, 35 ℃ and 37 ℃, and the influences of different fermentation temperatures on the viable count, the output of coenzyme Q10 and sensory score are compared. The results are shown in FIG. 5.
As can be seen from FIG. 5, the viable cell count of the cells and the content of coenzyme Q10 were the highest at 35 ℃ and reached 4.05X 107CFU/mL, coenzyme Q10 reached 5.33 mg/mL. The difference from other temperatures is very significant (p)<0.05), the sensory score had a score of 67.23 highest and 54.25 lowest at a temperature of 25 ℃. The optimal temperature is selected to be 35 ℃ in comprehensive consideration.
Influence of table rotation speed on viable count, coenzyme Q10 yield and sensory score
Measuring 100mL of apple juice, filling the apple juice into a 250mL shake flask, sterilizing the apple juice for 15min at 105 ℃, inoculating 10mL of rhodobacter sphaeroides target bacterial liquid, fermenting the apple juice at the fermentation temperature of 35 ℃, and comparing the influences of different table rotation speeds on the viable count, the coenzyme Q10 yield and sensory score, wherein the table rotation speeds are 100r/min, 120r/min, 150r/min, 180r/min and 200 r/min. The results are shown in FIG. 6.
As can be seen from FIG. 6, when the rotation speed of the shaking table is 180r/min, the maximum viable count of the cells is 5.13X 107CFU/mL, the highest coenzyme Q10 content is 10.45mg/L when the rotation speed is 200r/min, and the highest sensory score is 72.16 when the rotation speed of the shaking table is 100 r/min. Considering that the error of the sensory scoring is larger, the rotation speeds of 150r/min, 180r/min and 200r/min are finally selected for orthogonal optimization.
(6) Orthogonal optimization experiment
The single factor test determines that 4 factors which can obviously influence the viable count of the thallus, the content of coenzyme Q10 and the sensory score are respectively the inoculation amount, the liquid loading amount, the fermentation temperature and the rotating speed of a shaking table. Considering the significance of the temperature influence, the temperature is directly determined to be 35 ℃, and orthogonal optimization is carried out on the seed amount, the liquid loading amount and the rotating speed of the shaking table.
TABLE 1 orthogonal factor horizon
And (5) orthogonal optimization of test results and analysis.
TABLE 2 results and analysis of orthogonal experiments
TABLE 3 analysis of variance table of viable count
| Source of variation | Sum of squares of deviation | Degree of freedom | F ratio | Critical value of F | Significance of |
| Inoculum size (mL) | 908.809 | 2 | 439.250 | 19.00 | * |
| Liquid loading amount (mL) | 108.242 | 2 | 52.316 | 19.00 | * |
| Shaking table rotating speed (r/min) | 3764.536 | 2 | 1819.495 | 19.00 | * |
| Error of the measurement | 2.07 | 2 |
TABLE 4 analysis of variance table of coenzyme Q10
| Source of variation | Sum of squares of deviation | Degree of freedom | F ratio | Critical value of F | Significance of |
| Inoculum size (mL) | 14.812 | 2 | 11.403 | 19.00 | |
| Liquid loading amount (mL) | 3.100 | 2 | 2.386 | 19.00 | |
| Shaking table rotating speed (r/min) | 38.851 | 2 | 29.908 | 19.00 | * |
| Error of the measurement | 1.30 | 2 |
TABLE 5 ANOVA TABLE OF SENSORY SCORE
Note: denotes p <0.05
As can be seen from Table 2, the order of the factors affecting the viable count is the rotating speed of the rocking bed>Amount of inoculation>The liquid loading amount, as can be seen from table 3, these 3 factors have a significant effect on the viable cell count. Wherein the optimal level is that when the inoculation amount is 10mL, the liquid loading amount is 90mL, and the rotating speed of the shaking table is 180r/min, the viable count reaches 8.63 +/-0.1135 multiplied by 107CFU/mL。
Table 4 shows that the influence of each factor on coenzyme Q10 is in the order of table rotation speed > inoculum size > liquid loading, and that the influence of table rotation speed on coenzyme Q10 is significant, and the influence of inoculum size and liquid loading on coenzyme Q10 is not significant. Wherein the optimal level is that when the inoculation amount is 10mL, the liquid loading amount is 10mL, and the rotating speed of the shaking table is 200r/min, the content of the coenzyme Q10 reaches 9.83 +/-0.23 mg/L and is also high when the inoculation amount is 10mL, the liquid loading amount is 90mL, and the rotating speed of the shaking table is 180 r/min.
The influence sequence of each factor on the sensory score is the table rotation speed > liquid loading > inoculation amount, and as can be seen from table 5, the 3 factors have no significant influence on the sensory score. Wherein the sensory score is the optimal level when the inoculation amount is 11mL, the liquid loading amount is 110mL, and the rotating speed of the shaking table is 150 r/min.
In conclusion, due to the fact that the optimization conditions analyzed by each index independently are inconsistent, the major and minor influences and the influence significance of the factors are considered, and the inoculation amount of 10mL, the liquid loading amount of 90mL and the rotating speed of the shaking table of 180r/min are determined to be the optimal level of the fermentation process.
Table 6 shows the results of the test
| Sample numbering | Viable count (10)7CFU/mL) | Coenzyme Q10 production (m)g/L) | |
| 1 | 7.9 | 9.21 | 54.15 |
| 2 | 8.8 | 10.05 | 66.25 |
| 3 | 9.4 | 9.35 | 69.15 |
| Average | 8.7 | 9.54 | 63.18 |
To verify the accuracy of the optimized level, fermentation experiments were performed under optimized conditions and the results are shown in table 6. The results in Table 6 show that the number of viable bacteria obtained under the optimized conditions is 8.7X 106The yield of CFU/mL and coenzyme Q10 reaches 9.54mg/L and the sensory score is very close to the average value of a verification test, so that the influence of various screening factors on the viable count of rhodobacter sphaeroides, the yield of coenzyme Q10 and the sensory score of fermented fruit juice can be reflected more truly under the optimized condition, and the method has stronger guiding significance for the research of fermentation conditions.
(7) Fermentation comparative test of different fruit juices under optimized conditions
Under the preferable conditions of the above process, the number of viable bacteria and the output of coenzyme Q10 were determined after fermentation by comparing rhodobacter sphaeroides fermented pear juice and grape juice, as shown in Table 7.
TABLE 7 comparative fermentation tests of different juices
As can be seen from Table 7, the difference between the viable count and the coenzyme Q10 yield obtained after different fruit juices are fermented by rhodobacter sphaeroides is large, the viable count and the Q10 yield of apple juice are obviously higher than those of pear juice and grape juice, which indicates that the apple juice is suitable for rhodobacter sphaeroides fermentation, and the coenzyme Q10 content is higher.
(8) Measurement of physical and chemical indexes of fermented apple juice
The results of measuring the pH, total acid, SSC, total sugar, total phenol, and antioxidant ability of the fermented apple juice are shown in FIGS. 7 to 10.
As can be seen from fig. 7, the pH value increased and the total acid decreased as a whole in the fruit juice fermentation process. Rhodobacter sphaeroides belongs to non-oxygen-producing photosynthetic bacteria, various carbon sources such as organic acid and the like can be utilized, and the increase of the pH value is related to the metabolic activity of the rhodobacter sphaeroides.
The trend of the Soluble Solids Content (SSC) versus total sugar is shown in fig. 8, and it can be seen that SSC and total sugar content fluctuate greatly during fermentation, but decrease overall. Fermenting for 0-108 h, wherein SSC is reduced from 12.0 degrees Brix to 10.0 degrees Brix, and total sugar is reduced from 8.08g/100mL to 6.84g/100 mL. SSC is always in a descending trend before fermentation for 48 hours, the descending speed after fermentation for 36 hours is higher than that before fermentation for 36 hours, the ascending trend is carried out for 48-60 hours, the descending speed after fermentation for 60 hours begins, and the change point of total sugar is basically consistent with that of SSC mainly due to the biotransformation function of rhodobacter sphaeroides.
The trend of the total phenol content is shown in fig. 9, which shows that the total phenol content has a large variation range in the fermentation process, because the macromolecular phenols are decomposed into small molecular phenols under the action of microorganisms, which causes the total phenol content to vary.
The change in antioxidant properties during the fermentation of apple juice is shown in FIG. 10, expressed as DPPH clearance. From fig. 10, it can be seen that the DPPH clearance rate is highest up to 46.67% when the rhodobacter sphaeroides ferments the apple juice for 96 hours, and the DPPH clearance rate of the apple juice is 31.38%. According to the research, the apple juice fermented by rhodobacter sphaeroides has greatly improved oxidation resistance compared with the apple juice.
Example 1:
(1) taking a rhodobacter sphaeroides activation culture medium preserved by glycerol for static culture, wherein the culture temperature is 30-35 ℃, and the culture time is 48 h;
(2) seed culture solution: activated rhodobacter sphaeroides for 48 hours is transferred into a 250mL conical flask filled with 100mL of apple juice culture medium under the following culture conditions: 24h at 30-35 ℃ and 150 r/min;
(3) fermenting the apple juice: and (3) filling 90mL of fresh apple juice into a 250mL conical flask, carrying out autoclaving at 105 ℃ for 15min, adding the rhodobacter sphaeroides target bacterial liquid into the apple juice, and fermenting, wherein the addition amount of the rhodobacter sphaeroides is 10%, the fermentation temperature is 35 ℃, and the rotating speed of a shaking table is 180 r/min. Fermenting for 72h under the fermentation condition.
The viable count of rhodobacter sphaeroides in the fermented apple juice obtained by the embodiment reaches 8.6 multiplied by 107CFU/mL and the content of coenzyme Q10 reach 9.83 mg/L.
Example 2:
(1) taking a rhodobacter sphaeroides activation culture medium preserved by glycerol for static culture, wherein the culture temperature is 35 ℃, and the culture time is 48 hours;
(2) and (3) secondary activation: activated rhodobacter sphaeroides for 48 hours was transferred into a 250mL conical flask containing 100mL of activation medium under the following culture conditions: 24h at 30 ℃ and 150 r/min;
(3) fermenting the apple juice: putting 90mL of fresh apple juice into a 250mL conical flask, sterilizing at 105 ℃ for 15min under high pressure, adding the rhodobacter sphaeroides target bacterial liquid into the apple juice, fermenting, wherein the addition amount of the rhodobacter sphaeroides is 10%, the fermentation temperature is 30 ℃, and the rotating speed of a shaking table is 180 r/min. Fermenting for 72h under the fermentation condition.
The viable count of rhodobacter sphaeroides in the fermented apple juice obtained by the embodiment reaches 8.7107CFU/mL and coenzyme Q10 content reach 9.51 mg/L.
From the results, the high-content coenzyme Q10 fermented apple juice prepared by the invention is rich in rhodobacter sphaeroides and coenzyme Q10.
Claims (2)
1. A fermentation production method of apple juice rich in coenzyme Q10 is characterized in that the fermentation raw material in the fermentation production method is apple juice, the fermentation strain is preserved in China general microbiological culture collection center, and the fermentation strain is rhodobacter sphaeroides with the preservation number of CGMCC No 1.2182;
according to the volume of the apple juice, the inoculation amount of rhodobacter sphaeroides seed bacterial liquid is 10 percent;
the preparation of rhodobacter sphaeroides seed bacterial liquid comprises the following steps:
a. and (3) strain activation culture: taking 200uL of rhodobacter sphaeroides from a glycerol tube, and standing and culturing the rhodobacter sphaeroides in an activation culture medium at the temperature of 30 ℃ for 48 hours to prepare an activation 1-generation culture solution of rhodobacter sphaeroides;
b. seed liquid culture: according to the volume of the apple juice, 10 percent of activated 1-generation culture solution is added into an apple juice culture medium, and dark culture is carried out for 48 hours in a shaking table at 30 ℃ and 150r/min to obtain rhodobacter sphaeroides seed bacterial solution;
the activation medium is as follows: according to the addition amount of each liter of water, 20g/L of glucose, 10g/L of yeast extract, 10g/L of peptone, 0.05g/L of magnesium sulfate, 0.05g/L of monopotassium phosphate and 0.05g/L of dipotassium phosphate;
the fermentation temperature of the fermentation is 35 ℃, the rotating speed of a shaking table is 180r/min, and the fermentation time is 72 h;
the preparation of the apple juice comprises the following steps: juicing the apple blocks, adding 450mg/L pectinase for enzymolysis at 45 ℃ for 2h, and filtering to obtain apple juice.
2. A coenzyme Q10 apple juice, wherein the coenzyme Q10 apple juice is prepared by the method of claim 1.
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| CN102771871A (en) * | 2012-06-07 | 2012-11-14 | 山东省烟台农业学校 | Preparation of photosynthetic bacteria functional fruit juice beverage |
| CN108094790A (en) * | 2017-12-22 | 2018-06-01 | 光明乳业股份有限公司 | A kind of fermented apple juice and preparation method thereof |
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| CN1393546A (en) * | 2001-06-26 | 2003-01-29 | 山东万和生物科技有限公司 | Industrial fermenting process of sphaeroid red bacteria and its fermented liquid |
| CN102771871A (en) * | 2012-06-07 | 2012-11-14 | 山东省烟台农业学校 | Preparation of photosynthetic bacteria functional fruit juice beverage |
| CN108094790A (en) * | 2017-12-22 | 2018-06-01 | 光明乳业股份有限公司 | A kind of fermented apple juice and preparation method thereof |
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