Method for Increasing Content of Mogroside V in Siraitia Grosvenorii Suspended Cells

Abstract

The present invention relates to a method for increasing the content of mogroside V in Siraitia grosvenorii suspended cells. In the method, 10-200 mg/L of a yeast inducer and 50-260 mg/L of methyl jasmonate are respectively added on days 5 to 11 of the culture cycle of Siraitia grosvenorii suspended cells, thereby increasing the synthesis rate and yield of mogroside V, and also reducing the amount of mogroside II secreted. The method is beneficial for the market-oriented and large-scale production and quality control management of mogroside V.

Claims

1. A method for increasing content of mogroside V in Siraitia grosvenorii suspended cells, wherein the method comprises: adding a yeast inducer and methyl jasmonate during culturing of Siraitia grosvenorii suspended cells in a primary Siraitia grosvenorii suspended cell system.

2. The method according to claim 1, wherein during the culturing of the Siraitia grosvenorii suspended cells in the primary Siraitia grosvenorii suspended cell system, 10-50 mg/L of the yeast inducer and 50-100 mol/L of methyl jasmonate are added on days 5 to 7, 100-200 mg/L of the yeast inducer and 120-260 mol/L of methyl jasmonate are added on days 9 to 11, and the mature Siraitia grosvenorii suspended cells are harvested on days 17 to 25.

3. The method according to claim 1, wherein during the culturing of the mature Siraitia grosvenorii suspended cells in the primary Siraitia grosvenorii suspended cell system, 50 mg/L of the yeast inducer and 100 mol/L of methyl jasmonate are added on day 7, 180 mg/L of the yeast inducer and 220 mol/L of methyl jasmonate are added on day 11, and the mature Siraitia grosvenorii suspended cells are harvested on day 19.

4. The method according to claim 1, wherein the primary Siraitia grosvenorii suspended cell system is obtained by the following steps: (1) taking mature Siraitia grosvenorii embryos, subculturing on an MS semi-solid medium containing 30 g/L of sucrose, 1.0 mg/L of 6-benzylamino adenine, 0.5 mg/L of naphthaleneacetic acid, 100 mg/L of inositol and 4.6 g/L of agar in a pH range of 5.5-6.0, and selecting fresh embryogenic calli that are successively subcultured 3-5 times, that grow vigorously, and that have loose texture and a stable and uniform state; (2) transferring the fresh embryogenic calli obtained in step (1) at an inoculum amount of 100 g/L to an MS liquid medium containing 30 g/L of sucrose, 1.0 mg/L of 6-benzylamino adenine, 0.5 mg/L of naphthaleneacetic acid and 100 mg/L of inositol in a pH range of 5.5-6.5, and culturing in a shaker at a rotation speed of 150 r/min, a culture temperature of (252) C., a light intensity of 4000 lux and an illumination time of 12 h/d, to the primary Siraitia grosvenorii suspended cell system.

5. The method according to claim 4, wherein the subculture is performed in the step (1) for 4 times.

6. A method for preparing the mogroside V, wherein the mature Siraitia grosvenorii suspended cells prepared by the method according to 1 are further extracted and isolated to prepare the mogroside V.

7. A method for preparing the mogroside V, wherein the mature Siraitia grosvenorii suspended cells prepared according to claim 4 are further extracted and isolated to prepare the mogroside V.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0025] The following examples are intended to illustrate the present invention, but are not intended to limit the scope thereof.

Example 1

[0026] (1) Taking mature Siraitia grosvenorii embryos, subculturing on an MS semi-solid medium containing 30 g/L of sucrose, 1.0 mg/L of 6-benzylamino adenine, 0.5 mg/L of naphthaleneacetic acid, 100 mg/L of inositol and 4.6 g/L of agar in a pH range of 5.5-6.0, and selecting fresh embryogenic calli that were successively subcultured for 4 times, grew vigorously, and had loose texture and a stable and uniform state;

[0027] (2) the fresh embryogenic calli obtained in the step (1) were transferred at an inoculum amount of 100 g/L to an MS liquid medium containing 30 g/L of sucrose, 1.0 mg/L of 6-benzylamino adenine, 0.5 mg/L of naphthaleneacetic acid and 100 mg/L of inositol in a pH range of 5.5-6.5, and cultured in a shaker at a rotation speed of 150 r/min, a culture temperature of (252) C., a light intensity of 4000 lux and an illumination time of 12 h/d, so that the primary Siraitia grosvenorii suspended cell system was obtained;

[0028] (3) the mature Siraitia grosvenorii suspended cells were cultured in the primary Siraitia grosvenorii suspended cell system obtained in the step (2); 50 mg/L of a yeast inducer and 100 mol/L of methyl jasmonate were added on day 7 of the culture, and 180 mg/L of the yeast inducer and 220 mol/L of methyl jasmonate were added on day 11; and

[0029] (4) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Example 2

[0030] (1) Taking mature Siraitia grosvenorii embryos, subculturing on an MS semi-solid medium containing 30 g/L of sucrose, 1.0 mg/L of 6-benzylamino adenine, 0.5 mg/L of naphthaleneacetic acid, 100 mg/L of inositol and 4.6 g/L of agar in a pH range of 5.5-6.0, and selecting fresh embryogenic calli that were successively subcultured 3 times, grew vigorously, and had loose texture and a stable and uniform state;

[0031] (2) the fresh embryogenic calli obtained in the step (1) were transferred at an inoculum amount of 100 g/L (fresh weight) to an MS liquid medium containing 30 g/L of sucrose, 1.0 mg/L of 6-benzylamino adenine, 0.5 mg/L of naphthaleneacetic acid and 100 mg/L of inositol in a pH range of 5.5-6.5, and cultured in a shaker at a rotation speed of 150 r/min, a culture temperature of (252) C., a light intensity of 4000 lux and an illumination time of 12 h/d, so that the primary Siraitia grosvenorii suspended cell system was obtained;

[0032] (3) the mature Siraitia grosvenorii suspended cells were cultured in the primary Siraitia grosvenorii suspended cell system obtained in the step (2); 10 mg/L of a yeast inducer and 50 mol/L of methyl jasmonate were added on day 5 of the culture, and 100 mg/L of the yeast inducer and 120 mol/L of methyl jasmonate were added on day 9; and

[0033] (4) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Example 3

[0034] (1) Taking mature Siraitia grosvenorii embryos, subculturing on an MS semi-solid medium containing 30 g/L of sucrose, 1.0 mg/L of 6-benzylamino adenine, 0.5 mg/L of naphthaleneacetic acid, 100 mg/L of inositol and 4.6 g/L of agar in a pH range of 5.5-6.0, and selecting fresh embryogenic calli that were successively subcultured 5 times, grew vigorously, and had loose texture and a stable and uniform state;

[0035] (2) the fresh embryogenic calli obtained in the step (1) were transferred at an inoculum amount of 100 g/L (fresh weight) to an MS liquid medium containing 30 g/L of sucrose, 1.0 mg/L of 6-benzylamino adenine, 0.5 mg/L of naphthaleneacetic acid and 100 mg/L of inositol in a pH range of 5.5-6.5, and cultured in a shaker at a rotation speed of 150 r/min, a culture temperature of (252) C., a light intensity of 4000 lux and an illumination time of 12 h/d, so that the primary Siraitia grosvenorii suspended cell system was obtained;

[0036] (3) the mature Siraitia grosvenorii suspended cells were cultured in the primary Siraitia grosvenorii suspended cell system obtained in the step (2); 25 mg/L of a yeast inducer and 75 mol/L of methyl jasmonate were added on day 7 of the culture, and 200 mg/L of the yeast inducer and 260 mol/L of methyl jasmonate were added on day 11; and

[0037] (4) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Example 4

[0038] (1) Inoculating with and inducing Siraitia grosvenorii stem calli, subculturing on an MS medium containing 2.5% of sucrose, 0.2 mg/L of 6-benzyladenine, 0.04 mg/L of naphthylacetic acid and 4.0 mg/L of agar at a pH of 5.8, and selecting fresh, soft and fragile yellow-white calli that were successively subcultured 3, 4 or 5 times;

[0039] (2) the calli obtained in the step (1) were transferred at an inoculum amount of 300 g/l to the MS liquid medium in a sterile triangular flask containing 4.0% of sucrose, 1.0 mg/L of 6-benzylamino adenine, and 0.02 mg/L of naphthalene acetic acid, with the pH of the initial medium at 5.5 to 6.0; after suspension culture in a shaking incubator at 120 r/min in the dark at 25 C. for 24 h, the medium was left to stand, and the upper medium together with the single cells and small cell clusters was transferred to a sterile empty triangular flask to proceed with shaking culture;

[0040] (3) after 3 days, the same method was used to transfer once again; after 7 days, the suspended cells were filtered through a 40 m sieve, and inoculated into a fresh medium to proceed with the shaking culture; after reaching a certain volume, the suspended cells were transferred to a large triangular flask to proceed with the shaking culture with the addition of medium in the same way, until a certain amount of uniform and stable cell suspension culture fluid was reached; the cell suspension culture temperature was (252) C., the light intensity was 2000 lux, and the illumination time was 12 h/d; the primary Siraitia grosvenorii suspended cell system was thus obtained;

[0041] (4) the mature Siraitia grosvenorii suspended cells were cultured in the primary Siraitia grosvenorii suspended cell system obtained in the step (3); a yeast inducer of 50 mg/L and methyl jasmonate of 100 mol/L were added on day 7 of the culture, and the yeast inducer 180 mg/L and methyl jasmonate of 220 mol/L were added on day 11; and

[0042] (5) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Comparative Example 1

[0043] This comparative example was used to evaluate the difference in technical effects between the technical solution of adding an Aspergillus niger inducer and methyl jasmonate and the technical solution of the present invention. The specific steps were as follows:

[0044] (1) Taking mature Siraitia grosvenorii embryos, subculturing on an MS semi-solid medium containing sucrose of 30 g/L, 6-benzylamino adenine of 1.0 mg/L, naphthaleneacetic acid of 0.5 mg/L, inositol of 100 mg/L and agar of 4.6 g/L in a pH range of 5.5-6.0, and selecting fresh embryogenic calli that were successively subcultured for 4 times, grew vigorously, and had loose texture and a stable and uniform state;

[0045] (2) the fresh embryogenic calli obtained in the step (1) were transferred at an inoculum amount of 100 g/L to an MS liquid medium containing sucrose of 30 g/L, 6-benzylamino adenine of 1.0 mg/L, naphthaleneacetic acid of 0.5 mg/L and inositol of 100 mg/L in a pH range of 5.5-6.5, and cultured in a shaker at a rotation speed of 150 r/min, a culture temperature of (252) C., a light intensity of 4000 lux and an illumination time of 12 h/d, so that the primary Siraitia grosvenorii suspended cell system was obtained;

[0046] (3) the mature Siraitia grosvenorii suspended cells were cultured in the primary Siraitia grosvenorii suspended cell system obtained in the step (2); an Aspergillus niger inducer of 50 mg/L and methyl jasmonate of 100 mol/L were added on day 7 of the culture, and the Aspergillus niger inducer of 180 mg/L and methyl jasmonate of 220 mol/L were added on day 11; and

[0047] (4) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Comparative Example 2

[0048] This comparative example was used to evaluate the difference in technical effects between the technical solution of adding phenylalanine and a yeast inducer and the technical solution of the present invention. The specific steps were as follows:

[0049] (1) Taking mature Siraitia grosvenorii embryos, subculturing on an MS semi-solid medium containing sucrose of 30 g/L, 6-benzylamino adenine of 1.0 mg/L, naphthaleneacetic acid of 0.5 mg/L, inositol of 100 mg/L and agar of 4.6 g/L in a pH range of 5.5-6.0, and selecting fresh embryogenic calli that were successively subcultured for 4 times, grew vigorously, and had loose texture and a stable and uniform state;

[0050] (2) the fresh embryogenic calli obtained in the step (1) were transferred at an inoculum amount of 100 g/L to an MS liquid medium containing sucrose of 30 g/L, 6-benzylamino adenine of 1.0 mg/L, naphthaleneacetic acid of 0.5 mg/L and inositol of 100 mg/L in a pH range of 5.5-6.5, and cultured in a shaker at a rotation speed of 150 r/min, a culture temperature of (252) C., a light intensity of 4000 lux and an illumination time of 12 h/d, so that the primary Siraitia grosvenorii suspended cell system was obtained;

[0051] (3) the mature Siraitia grosvenorii suspended cells were cultured in the primary Siraitia grosvenorii suspended cell system obtained in the step (2); phenylalanine of 50 mg/L and the yeast inducer of 50 mg/L were added on day 7 of the culture, and phenylalanine of 180 mg/L and the yeast inducer of 180 mg/L were added on day 11 of the culture; and

[0052] (4) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Comparative Example 3

[0053] This comparative example was used to evaluate the difference in technical effects between the technical solution of adding the yeast inducer alone and the technical solution of the present invention. The specific steps were as follows:

[0054] (1) Taking mature Siraitia grosvenorii embryos, subculturing on an MS semi-solid medium containing sucrose of 30 g/L, 6-benzylamino adenine of 1.0 mg/L, naphthaleneacetic acid of 0.5 mg/L, inositol of 100 mg/L and agar of 4.6 g/L in a pH range of 5.5-6.0, and selecting fresh embryogenic calli that were successively subcultured for 4 times, grew vigorously, and had loose texture and a stable and uniform state;

[0055] (2) the fresh embryogenic calli obtained in the step (1) were transferred at an inoculum amount of 100 g/L to an MS liquid medium containing sucrose of 30 g/L, 6-benzylamino adenine of 1.0 mg/L, naphthaleneacetic acid of 0.5 mg/L and inositol of 100 mg/L in a pH range of 5.5-6.5, and cultured in a shaker at a rotation speed of 150 r/min, a culture temperature of (252) C., a light intensity of 4000 lux and an illumination time of 12 h/d, so that the primary Siraitia grosvenorii suspended cell system was obtained;

[0056] (3) the mature Siraitia grosvenorii suspended cells were cultured in the primary Siraitia grosvenorii suspended cell system obtained in the step (2); the yeast inducer of 50 mg/L was added on day 7 of the culture, and the yeast inducer of 180 mg/L was added on day 11; and

[0057] (4) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Comparative Example 4

[0058] This comparative example was used to evaluate the difference in technical effects between the technical solution of adding methyl jasmonate alone and the technical solution of the present invention. The specific steps were as follows:

[0059] (1) Taking mature Siraitia grosvenorii embryos, subculturing on an MS semi-solid medium containing sucrose of 30 g/L, 6-benzylamino adenine of 1.0 mg/L, naphthaleneacetic acid of 0.5 mg/L, inositol of 100 mg/L and agar of 4.6 g/L in a pH range of 5.5-6.0, and selecting fresh embryogenic calli that were successively subcultured for 4 times, grew vigorously, and had loose texture and a stable and uniform state;

[0060] (2) the fresh embryogenic calli obtained in the step (1) were transferred at an inoculum amount of 100 g/L to an MS liquid medium containing sucrose of 30 g/L, 6-benzylamino adenine of 1.0 mg/L, naphthaleneacetic acid of 0.5 mg/L and inositol of 100 mg/L in a pH range of 5.5-6.5, and cultured in a shaker at a rotation speed of 150 r/min, a culture temperature of (252) C., a light intensity of 4000 lux and an illumination time of 12 h/d, so that the primary Siraitia grosvenorii suspended cell system was obtained;

[0061] (3) the mature Siraitia grosvenorii suspended cells were cultured in the primary Siraitia grosvenorii suspended cell system obtained in the step (2); methyl jasmonate of 100 mol/L was added on day 7 of the culture, and methyl jasmonate of 220 mol/L was added on day 11; and

[0062] (4) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Comparative Example 5

[0063] This comparative example was used to evaluate the difference in technical effects between the technical solution disclosed in Reference 1 and the technical solution of the present invention. The specific steps were as follows:

[0064] (1) Inoculating with and inducing Siraitia grosvenorii stem calli, subculturing on an MS medium containing 2.5% of sucrose, 0.2 mg/L of 6-benzyladenine, 0.04 mg/L of naphthylacetic acid and 4.0 mg/L of agar at a pH of 5.8, and selecting fresh, soft and fragile yellow-white calli that were successively subcultured 3, 4 or 5 times;

[0065] (2) the calli obtained in the step (1) were transferred at an inoculum amount of 300 g/l to the MS liquid medium in a sterile triangular flask containing 4.0% of sucrose, 1.0 mg/L of 6-benzylamino adenine, and 0.02 mg/L of naphthalene acetic acid, with the pH of the initial medium at 5.5 to 6.0; after suspension culture in a shaking incubator at 120 r/min in the dark at 25 C. for 24 h, the medium was left to stand, and the upper medium together with the single cells and small cell clusters was transferred to a sterile empty triangular flask to proceed with shaking culture;

[0066] (3) after 3 days, the same method was used to transfer once again; after 7 days, the suspended cells were filtered through a 40 m sieve, and inoculated into a fresh medium to proceed with the shaking culture; after reaching a certain volume, the suspended cells were transferred to a large triangular flask to proceed with the shaking culture with the addition of medium in the same way, until a certain amount of uniform and stable cell suspension culture fluid was reached; the cell suspension culture temperature was (252) C., the light intensity was 2000 lux, and the illumination time was 12 h/d; the primary Siraitia grosvenorii suspended cell system was thus obtained; and

[0067] (4) on days 1 to 31 of the culture, the Siraitia grosvenorii suspended cells were extracted every other day to detect the content of mogroside V and mogroside II.

Comparison of Contents of Mogroside V

1. Experimental Method

[0068] Weighing 1.5 g Siraitia grosvenorii dry weight cells, adding 60% ethanol at a feed-to-liquid ratio of 15:1, ultrasonically extracting for 40 min, extracting for 2 h at 90 C. in a slightly boiling state, repeating the extraction for 3 times, and combining the extracts to concentrate them to 5 mL in a rotary evaporator. The concentrate was filtered through a 0.22 m filtration membrane, and then qualitative and quantitative analysis was performed using a high-performance liquid chromatograph.

[0069] Chromatographic conditions: The chromatographic column was an inverted C18 column, 4.6 mm250 mm; the mobile phase was water-acetonitrile at a ratio of 78:22, the flow rate was 1.0 mL/min; and the detection wavelength was 203 nm.

[0070] 2. Results

[0071] The effect of different treatment conditions on the yield of mogroside V per unit time is shown in Table 1.

TABLE-US-00001 TABLE 1 The effect of different treatment conditions on the yield of mogroside V per unit time (n = 5; x s; g/L) Comparative Comparative Comparative Comparative Comparative Example 1 Example 1 Example 2 Example 3 Example 4 Example 5 1 d 31.1 0.8 31.4 0.6 32.6 0.5 31.5 0.5 31.1 0.8 30.0 1.0 3 d 32.6 1.2 31.9 1.4 33.0 1.6 31.8 2.1 32.6 0.9 31.6 0.6 5 d 33.6 1.1 32.9 1.2 33.2 1.4 32.8 1.8 33.2 1.1 32.2 0.8 7 d 34.2 1.4 33.2 1.1 34.2 1.8 33.0 1.6 33.5 1.5 33.6 1.2 9 d 43.5 2.1 33.8 1.3 35.6 1.5 33.6 1.4 39.5 2.0 39.8 1.5 11 d 52.6 1.8 34.2 1.6 36.8 1.6 34.0 1.2 46.5 1.8 45.5 1.7 13 d 64.8 2.0 34.7 1.5 41.5 2.0 34.5 2.2 51.6 1.8 50.5 2.2 15 d 73.8 2.8 35.2 2.2 47.5 2.6 36.8 1.6 53.6 1.5 56.4 2.4 17 d 79.9 2.6 36.2 2.1 53.4 2.2 43.5 1.8 54.5 1.6 60.8 2.8 19 d 85.2 2.5 37.8 2.4 60.8 2.1 49.5 2.0 55.7 1.8 67.5 2.5 21 d 79.5 2.6 39.4 2.5 67.2 2.3 55.4 2.2 56.8 2.0 70.7 3.1 23 d 78.6 2.8 52.6 3.0 72.0 2.8 61.8 2.6 58.2 2.2 69.2 3.2 25 d 76.5 2.5 64.8 2.8 75.6 2.6 68.5 2.5 55.2 2.4 65.8 3.0 27 d 74.2 2.6 73.8 2.5 77.2 2.8 71.8 3.0 51.4 2.5 63.6 2.8 29 d 72.1 3.1 75.9 2.9 79.0 2.5 72.2 2.7 46.5 2.4 59.4 3.2 31 d 68.2 2.8 76.2 2.6 78.8 2.5 72.0 2.8 41.2 2.5 55.6 2.8

[0072] The effect of different treatment conditions on the peak yield (the maximum yield) of mogroside V is shown in Table 2.

TABLE-US-00002 TABLE 2 The effect of different treatment conditions on the peak yield (the maximum yield) of mogroside V (n = 5; x s; g/L) Peak yield (maximum yield) of mogroside V Example 1 85.2 2.5 Example 2 84.9 1.8 Example 3 86.7 2.2 Example 4 85.1 3.0 Comparative Example 1 76.2 2.6 Comparative Example 2 79.0 2.5 Comparative Example 3 72.2 2.7 Comparative Example 4 58.2 2.2 Comparative Example 5 70.7 3.1

Comparison of Contents of Mogroside II

1. Experimental Method

[0073] Weighing 1.5 g Siraitia grosvenorii dry weight cells, adding 60% ethanol at a feed to liquid ratio of 15:1, ultrasonically extracting for 40 min, extracting for 2 h at 90 C. in a slightly boiling state, repeating the extraction for 3 times, and combining the extracts to concentrate them to 5 mL in a rotary evaporator. The concentrate was filtered through a 0.22 m filtration membrane, and then qualitative and quantitative analysis was performed using a high-performance liquid chromatograph.

[0074] Chromatographic conditions: The chromatographic column was a ZORBAX SBC C18 column, 4.6 mm150 mm; the mobile phase was water-acetonitrile gradient elution (0-25 min, 13.5% to 35% acetonitrile), the flow rate was 0.8 mL/min; and the detection wavelength was 203 nm.

2. Results

[0075] The effect of different treatment conditions on the content of mogroside II at the peak of mogroside V is shown in Table 3.

TABLE-US-00003 TABLE 3 The effect of different treatment conditions on the content of mogroside II at the peak of mogroside V (n = 5; x s; g/L) Content of mogroside II Example 1 17.2 2.1 Example 2 17.6 2.0 Example 3 18.5 1.4 Example 4 18.3 2.6 Comparative Example 1 42.6 4.3 Comparative Example 2 39.8 3.8 Comparative Example 3 40.2 2.2 Comparative Example 4 32.8 2.6 Comparative Example 5 30.4 1.6

CONCLUSION

[0076] The results of comparison of the contents of mogroside V (Tables 1 and 2) show that the contents of mogroside V in Examples 1-4 were significantly higher than those of Comparative Examples 1-5, with the peak values at 85.2 g/L, 84.91.8 g/L, 86.72.2 g/L and 85.13.0 g/L, respectively, indicating that the addition of the yeast inducer and methyl jasmonate to the Siraitia grosvenorii suspended cell culture system could effectively increase the yield of mogroside V, with the technical effect obviously superior to the prior art. Besides, in comparison with Comparative Examples 1-5, the synthesis rate of mogroside V in Example 1 was significantly increased, with the content of mogroside V higher than 70 g/L on day 15 of the culture and reaching the peak value on day 19 of the culture. The results show that the addition of the yeast inducer and methyl jasmonate in the Siraitia grosvenorii suspended cell culture system could effectively increase the synthesis rate of mogroside V by the Siraitia grosvenorii suspended cells, with the technical effect obviously superior to the prior art.

[0077] The results of comparison of the contents of mogroside II (Table 3) show that the contents of mogroside II in Examples 1-5 were significantly reduced compared with Comparative Examples 1-5, indicating that the addition of the yeast inducer and methyl jasmonate to the Siraitia grosvenorii suspended cell culture system could effectively reduce the content of the bitter component mogroside II, with the technical effect obviously superior to the prior art.

[0078] Although the present invention has been described in detail by using general descriptions, specific embodiments and tests, some modifications or improvements can be made based on the present invention, which will be obvious to those skilled in the art. Therefore, these modifications or improvements made without departing from the spirit of the present invention shall all fall within the scope of protection of the present invention.

INDUSTRIAL PRACTICALITY

[0079] The present invention provides a method for increasing the content of mogroside V in Siraitia grosvenorii suspended cells. In the method, 10-200 mg/L of a yeast inducer and 50-260 mg/L of methyl jasmonate are respectively added on days 5 to 11 of the culture cycle of Siraitia grosvenorii suspended cells, thereby increasing the synthesis rate and yield of mogroside V, and also reducing the amount of mogroside II secreted. The method of the present invention, artificially regulating the production process of Siraitia grosvenorii cells and mogroside V, is beneficial for the market-oriented and large-scale production and quality control management of mogroside V, and has good economic value and broad application prospects.