Optimization of Fermentation Conditions and Product Identification of a Saponin-Producing Endophytic Fungus.
Microorganisms
BACKGROUND:Some fungal endophytes isolated from may present a new method of obtaining saponins. This experiment aimed to optimize the total saponin yield produced through in vitro fermentation by an endophytic fungus and analyze its saponin species in the fermented extract. METHODS:Fermentation protocols were optimized with a uniform design and verified through regression analysis to maximize the total saponin yield. The saponin types under optimal fermentation conditions were then identified and analyzed using Liquid Chromatography-Mass Spectrometry. RESULTS:The strain NSJ105 (gene accession number: OR144428) isolated from wild ginseng could produce total saponins. The total saponin yield could be increased more than two-fold through the optimization of fermentation conditions. The concentration of the total saponins achieved by the verified protocol 105-DP was close to the predicted value. The fermentation conditions of the 105-DP protocol were as follows: potato concentration 97.3 mg/mL, glucose concentration 20.6 mg/mL, inoculum volume 2.1%, fermentation broth pH 2.1, fermentation temperature 29.2 °C, and fermentation time 6 d. It was detected and analyzed that the fermented extract of 105-DP contained the ginsenosides Rf and Rb. CONCLUSION:The endophytic fungus strain NSJ105 has potential application value in saponin production.
10.3390/microorganisms11092331
Microbiome-based screening and co-fermentation of rhizospheric microorganisms for highly ginsenoside Rg production.
Microbiological research
Ginsenoside Rg has a wide range of pharmacological activities and application value while the content of Rg in Panax plants is extremely low. The interaction between medicinal plants and microorganisms will be beneficial to produce active compounds by biotechnology. In this study, the rhizosphere soil samples from different P. notoginseng producing areas (Asanlong, Huilong, Demonstration garden) were collected to analyze the soil microflora characteristics by Illumina MiSeq sequencing technology. Based on the highest contents of ginsenosides in Huilong, the first, second, and the specificity dominant strains were predicted and the database was established. Besides, a total of 6 strains of bacteria and 3 strains of fungi were isolated from the soil of P. notoginseng. Among them, F24 was identified as Chaetomium sp, which was not only consistent with the second dominant strain of P. notoginseng predicted in the database, but also had a high level of Rg. It also indicated that the method of screening the dominant strains in soil was reasonable and the database was reliable. Through the optimization of fermentation conditions, the highest yield of Rg was obtained when F24 was cultured in the medium supplemented with 8 mg/L glucose and 5 mg/L methyl jasmonate (MeJA) for 7 days. On this basis, the yield of ginsenoside Rg in shake flask reached 108.95 mg/L (4.93 mg/g, ~4.2-fold higher than cultivated P. notoginseng plants) by the co-fermentation with A. niger in a concentration ratio of 2:1. The increase of Rg yield mainly came from the conversion of other 20S-protopanaxdiol saponins into Rg by β-glucosidase in A. niger. In addition, the expression levels of ginsenoside biosynthesis genes in different strains were compared, it was found that the expression of key genes was significantly increased after co-fermentation. This research provided certain theoretical and technical support for the large-scale industrial production of ginsenoside Rg by microbiology.
10.1016/j.micres.2022.127054
High-density immobilization of a ginsenoside-transforming β-glucosidase for enhanced food-grade production of minor ginsenosides.
Cui Chang-Hao,Jeon Byeong-Min,Fu Yaoyao,Im Wan-Taek,Kim Sun-Chang
Applied microbiology and biotechnology
Use of recombinant glycosidases is a promising approach for the production of minor ginsenosides, e.g., Compound K (CK) and F, which have potential applications in the food industry. However, application of these recombinant enzymes for food-grade preparation of minor ginsenosides are limited by the lack of suitable expression hosts and low productivity. In this study, Corynebacterium glutamicum ATCC13032, a GRAS strain that has been used extensively for the industrial-grade production of additives for foodstuffs, was employed to express a novel β-glucosidase (MT619) from Microbacterium testaceum ATCC 15829 with high ginsenoside-transforming activity. A cellulose-binding module was additionally fused to the N-terminus of MT619 for immobilization on cellulose, which is an abundant and safe material. Via one-step immobilization, the fusion protein in cell lysates was efficiently immobilized on regenerated amorphous cellulose at a high density (maximum 984 mg/g cellulose), increasing the enzyme concentration by 286-fold. The concentrated and immobilized enzyme showed strong conversion activities against protopanaxadiol- and protopanaxatriol-type ginsenosides for the production of CK and F. Using gram-scale ginseng extracts as substrates, the immobilized enzyme produced 7.59 g/L CK and 9.42 g/L F in 24 h. To the best of our knowledge, these are the highest reported product concentrations of CK and F, and this is the first time that a recombinant enzyme has been immobilized on cellulose for the preparation of minor ginsenosides. This safe, convenient, and efficient production method could also be effectively exploited in the preparation of food-processing recombinant enzymes in the pharmaceutical, functional food, and cosmetics industries.
10.1007/s00253-019-09951-4
Application of microorganisms in Panax ginseng: cultivation of plants, and biotransformation and bioactivity of key component ginsenosides.
Archives of microbiology
Panax ginseng is a precious Chinese medicinal plant with a long growth cycle and high medicinal value. Therefore, it is of great significance to explore effective ways to increase its yield and main active substance content to reduce the cost of ginseng, which is widely used in food and clinical applications. Here, we review the key roles of microorganisms in the biological control of ginseng diseases, enhancement of ginseng yield, biotransformation of ginsenosides, and augmentation of ginsenoside bioactivity. The application of microorganisms in P. ginseng faces multiple challenges, including the need for further exploration of efficient microbial strain resources used in the cultivation of ginseng and biotransformation of ginsenosides, lack of microbial application in large-scale field cultivation of ginseng, and unclear mechanism of microbial transformation of ginsenosides. This review provides a deeper understanding of the applications of microorganisms in P. ginseng.
10.1007/s00203-024-04144-8
The anti-aging mechanism of ginsenosides with medicine and food homology.
Food & function
With the acceleration of global aging and the rise in living standards, the achievement of healthy aging is becoming an imperative issue globally. Ginseng, a medicinal plant that has a long history of dietary intake and remarkable medicinal value, has become a research hotspot in the field of food and medicine. Ginsenosides, especially protopanaxadiol-type saponins and protopanaxatriol-type saponins, are among the most important active ingredients in ginseng. Ginsenosides have been found to exhibit powerful and diverse pharmacological activities, such as antiaging, antitumor, antifatigue and immunity enhancement activities. Their effects in antiaging mainly include (1) promotion of metabolism and stem cell proliferation, (2) protection of skin and nerves, (3) modulation of intestinal flora, (4) maintenance of mitochondrial function, and (5) enhancement of telomerase activity. The underlying mechanisms are primarily associated with the intervention of the signaling pathways in apoptosis, inflammation and oxidative stress. In this review, the mechanism of action of ginsenosides in antiaging as well as the potential values of developing ginsenoside-based functional foods and antiaging drugs are discussed.
10.1039/d3fo02580b
Microbial Conversion of Protopanaxadiol-Type Ginsenosides by the Edible and Medicinal Mushroom : A Green Biotransformation Strategy.
Liu Zhi,Li Jia-Xin,Wang Chong-Zhi,Zhang Dan-Li,Wen Xin,Ruan Chang-Chun,Li Yu,Yuan Chun-Su
ACS omega
Previous studies have shown that many kinds of microorganisms, including bacteria, yeasts, and filamentous fungi, can convert parent ginsenosides into minor ginsenosides. However, most microorganisms used for ginsenoside transformations may not be safe for food consumption and drug development. In this study, 24 edible and medicinal mushrooms were screened by high-performance liquid chromatography analyses for their ability to microbiologically transform protopanaxadiol (PPD)-type ginsenosides. We observed that the degradation of ginsenosides by was inhibited by high concentrations of sugar in the culture medium. However, the inhibition was avoided by maintaining sugar concentration below 15 g L. showed a strong ability to convert PPD-type ginsenosides (Rb, Rc, Rb, and Rd) into minor ginsenosides (F, C-O, C-Y, C-Mc, C-Mc, and C-K). The production and bioconversion rates of minor ginsenosides were significantly higher than those previously reported by food microorganisms. The fermentation process is efficient, nontoxic, eco-friendly, and economical, and the required biotransformation systems are readily available. This is the first report about the biotransformation of major ginsenosides into minor ginsenosides through fermentation by edible and medicinal mushrooms. Our results provide a green biodegradation strategy in transformation of ginsenosides using edible and medicinal mushrooms.
10.1021/acsomega.9b01001
Changes in the Ginsenoside Content During Fermentation Using an Appliance for the Preparation of Red Ginseng.
Lee So Jin,Ha Na,Kim Yunjeong,Kim Min-Gul
The American journal of Chinese medicine
The total amount of ginsenoside in fermented red ginseng (FRG) is increased by microbial fermentation. The aim of this study was to evaluate whether fermentation time and temperature affect the ginsenoside content during fermentation using an appliance for the preparation of red ginseng. The FRG and fermented red ginseng extracts (FRG-e) were prepared using an appliance for the preparation of red ginseng. The temperature was recorded and time points for sampling were scheduled at pre-fermentation (0[Formula: see text]h) and 18, 36, 48, 60 and 72[Formula: see text]h after the addition of the microbial strains. Samples of FRG and FRG-e were collected to identify changes in the ginsenoside contents at each time point during the fermentation process. The ginsenoside content was analyzed using high performance liquid chromatography (HPLC). The levels of ginsenoside Rh1, Rg3, and compound Y, which are known to have effective pharmacological properties, increased more than three-fold in the final products of FRG relative to samples prior to fermentation. Although the ginsenoside constituents of FRG-e decreased or increased and then decreased during fermentation, the total amount of ginsenoside in FRG-e was even higher than those in FRG; the total amounts of ginsenoside in FRG-e and FRG were 8282.8 and 738.0[Formula: see text]mg, respectively. This study examined the changes in composition of ginsenosides and suggests a method to manufacture high-content total ginsenosides according to the fermentation temperature and process time. Reducing the extraction time is expected to improve the decrease of ginsenosides in FRG-e as a function of the fermentation time.
10.1142/S0192415X16500890
Ginsenoside Rk3 is a novel PI3K/AKT-targeting therapeutics agent that regulates autophagy and apoptosis in hepatocellular carcinoma.
Journal of pharmaceutical analysis
Hepatocellular carcinoma (HCC) is the third leading cause of cancer death worldwide. Ginsenoside Rk3, an important and rare saponin in heat-treated ginseng, is generated from Rg1 and has a smaller molecular weight. However, the anti-HCC efficacy and mechanisms of ginsenoside Rk3 have not yet been characterized. Here, we investigated the mechanism by which ginsenoside Rk3, a tetracyclic triterpenoid rare ginsenoside, inhibits the growth of HCC. We first explored the possible potential targets of Rk3 through network pharmacology. Both in vitro (HepG2 and HCC-LM3 cells) and in vivo (primary liver cancer mice and HCC-LM3 subcutaneous tumor-bearing mice) studies revealed that Rk3 significantly inhibits the proliferation of HCC. Meanwhile, Rk3 blocked the cell cycle in HCC at the G1 phase and induced autophagy and apoptosis in HCC. Further proteomics and siRNA experiments showed that Rk3 regulates the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway to inhibit HCC growth, which was validated by molecular docking and surface plasmon resonance. In conclusion, we report the discovery that ginsenoside Rk3 binds to PI3K/AKT and promotes autophagy and apoptosis in HCC. Our data strongly support the translation of ginsenoside Rk3 into novel PI3K/AKT-targeting therapeutics for HCC treatment with low toxic side effects.
10.1016/j.jpha.2023.03.006