Highly Selective Bioconversion of Ginsenoside Rb1 to Compound K by the Mycelium of Cordyceps sinensis under Optimized Conditions
Abstract
:1. Introduction
20(S)-Protopanaxdiol Type | ||||
Ginsenoside | R1 (C-3) | R2 (C-20) | ||
Rb1 | -Glc2-Glc | -Glc6-Glc | ||
Rd | -Glc2-Glc | -Glc | ||
Rg3 | -Glc2-Glc | -H | ||
F2 | -Glc | -Glc | ||
Rh2 | -Glc | -H | ||
Compound K | -H | -Glc | ||
20(S)-Protopanaxtriol Type | ||||
Ginsenoside | R1 (C-3) | R2 (C-6) | R3 (C-20) | |
Re | -H | -Glc2-Rha | -Glc | |
Rf | -H | -Glc2-Glc | -H | |
Rg1 | -H | -Glc | -Glc | |
Rg2 | -H | -Glc2-Rha | -H | |
Rh1 | -H | -Glc | -H |
2. Results and Discussion
2.1. Optimization on Biotransformation Parameters of Ginsenoside Rb1 by C. sinensis
2.1.1. Determination on Substrate-Feeding Time
2.1.2. Optimization on Carbon Sources, Nitrogen Sources, Elicitors, Reaction pH and Temperature for the Biotransformation Process
2.2. Biotransformation Pathway of Ginsenoside Rb1 by C. sinensis
2.3. Calculation on Molar Conversion Rate of Rb1 and Purity of CK
No. | Day 0 | Day 4 | ||
---|---|---|---|---|
Compound | MRM Intensity (×107) | Compound | MRM Intensity (×107) | |
1 | Rb1 | 2.3991 | Rb1 | - |
2 | Rg1 | 1.1094 | Rg1 | 1.1137 |
3 | CK | - | CK | 1.9749 |
No. | tR (min) | Identificaiton | Intensity | Relative Intensity (%) | Diff. (ppm) |
---|---|---|---|---|---|
1 | 18.117 | Rd | 215,429 | 3.1 | −1.02 |
2 | 20.185 | Unkown | 78,191 | 1.2 | - |
3 | 30.809 | F2 | 199,501 | 2.8 | −0.93 |
4 | 45.653 | CK | 6,331,023 | 91.4 | −0.61 |
5 | 51.291 | Unkown | 105,419 | 1.5 | - |
2.4. Identification of the Biotransformation Metabolites
Carbon Site | |||
---|---|---|---|
CK R1 = Glc, R2 = H | Metabolite (3) | Rh2 R1 = H, R2 = Glc | |
1 | 39.19 | 38.62 | 39.12 |
2 | 28.05 | 27.21 | 27.05 |
3 | 77.83 | 78.16 | 88.78 |
4 | 39.34 | 38.87 | 39.66 |
5 | 56.14 | 56.92 | 56.35 |
6 | 18.55 | 18.01 | 18.43 |
7 | 34.95 | 34.47 | 35.15 |
8 | 39.86 | 39.59 | 40.00 |
9 | 50.09 | 51.01 | 50.38 |
10 | 37.14 | 36.79 | 36.94 |
11 | 30.56 | 29.62 | 32.02 |
12 | 69.96 | 69.86 | 70.96 |
13 | 49.30 | 49.69 | 49.54 |
14 | 51.21 | 51.66 | 51.69 |
15 | 30.74 | 30.23 | 31.32 |
16 | 26.42 | 26.62 | 26.70 |
17 | 51.39 | 51.77 | 54.77 |
18 | 15.81 | 15.28 | 16.77 |
19 | 16.14 | 15.80 | 15.61 |
20 | 83.08 | 83.55 | 72.94 |
21 | 22.13 | 22.83 | 26.83 |
22 | 35.96 | 35.28 | 35.88 |
23 | 22.98 | 22.83 | 22.97 |
24 | 125.75 | 124.45 | 126.30 |
25 | 130.69 | 130.88 | 130.73 |
26 | 25.54 | 25.80 | 25.78 |
27 | 17.54 | 16.94 | 17.66 |
28 | 28.47 | 29.62 | 28.14 |
29 | 16.14 | 16.51 | 16.34 |
30 | 17.18 | 16.94 | 17.65 |
Glucosyl C-1″ | 98.10 | 96.92 | 106.92 |
3. Experimental Section
3.1. Materials and Chemicals
3.2. Microorganism and Culture Condition
3.3. Growth Curve of C. sinensis
3.4. Biotransformation of Ginsenoside Rb1 by C. sinensis
3.5. Sample Preparation
3.6. Thin Layer Chromatography Analysis
3.7. HPLC-Q-TOF-MS Analysis
3.8. Determination of the Metabolites
3.9. Parameters Optimization for the Biotransformation of Ginsenoside Rb1
3.9.1. Substrate-Feeding Time
3.9.2. Carbon Sources
3.9.3. Nitrogen Sources
3.9.4. Elicitors
3.9.5. Reaction pH
3.9.6. Biotransformation Temperature
3.10. Time-Course Experiments
- (1)
- Rb1: m/z 1107→945 [M − H]−
- (2)
- Rg1: m/z 799→637 [M − H]−
- (3)
- Rd: m/z 945→783 [M − H]−
- (4)
- F2: m/z 783→621 [M − H]−
- (5)
- CK: m/z 621→459 [M − H]−
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Wang, W.-N.; Yan, B.-X.; Xu, W.-D.; Qiu, Y.; Guo, Y.-L.; Qiu, Z.-D. Highly Selective Bioconversion of Ginsenoside Rb1 to Compound K by the Mycelium of Cordyceps sinensis under Optimized Conditions. Molecules 2015, 20, 19291-19309. https://doi.org/10.3390/molecules201019291
Wang W-N, Yan B-X, Xu W-D, Qiu Y, Guo Y-L, Qiu Z-D. Highly Selective Bioconversion of Ginsenoside Rb1 to Compound K by the Mycelium of Cordyceps sinensis under Optimized Conditions. Molecules. 2015; 20(10):19291-19309. https://doi.org/10.3390/molecules201019291
Chicago/Turabian StyleWang, Wei-Nan, Bing-Xiong Yan, Wen-Di Xu, Ye Qiu, Yun-Long Guo, and Zhi-Dong Qiu. 2015. "Highly Selective Bioconversion of Ginsenoside Rb1 to Compound K by the Mycelium of Cordyceps sinensis under Optimized Conditions" Molecules 20, no. 10: 19291-19309. https://doi.org/10.3390/molecules201019291