Ферментированные средства традиционной китайской медицины: от лабораторных инноваций до индустриализации – подробное руководство

Fermented traditional Chinese medicine (TCM) represents a brilliant fusion of ancient 炮制工艺 (processing techniques) and cutting-edge technology. For millennia, fermentation has been integral to preparing classic TCMs like 六神曲 (Liushenqu) and 淡豆豉 (Danshichi). Today, modern biotechnologies are revolutionizing this age-old practice, enhancing efficacy, standardization, and scalability. This article delves into key preparation cases, industrial applications, and future trends, uncovering the “magic transformation” of fermented TCM from lab to mass production.

Case 1: Liushenqu – Standardized Practice of Multi-Strain Synergistic Fermentation

Liushenqu, a classic fermented TCM, has long relied on natural fermentation, leading to unstable microbial communities and significant quality fluctuations. Modern advancements have addressed these issues through targeted microbial screening и systematic process optimization.

Key Preparation Steps

• Optimized Raw Material Ratio: The formula is precisely calibrated as almond: adzuki bean: flour: Artemisia annua juice: Polygonum hydropiper juice = 1:1:8:0.5:0.5 (mass ratio), with total carbon sources controlled at 65% to ensure optimal microbial growth.
• Three-Stage Fermentation Control:
  • Primary seed culture: 32°C, 180rpm, 48h (liquid fermentation for strain activation).
  • Secondary expansion culture: 28°C, solid-state fermentation for 72h (scale-up to adapt to solid matrix).
  • Tertiary industrial fermentation: 75% environmental humidity with intelligent pH regulation (ensuring stable metabolite production).
• Innovative Quality Evaluation: A “three-dimensional” quality control system is established, including:
  • Enzyme activity index (amylase ≥ 500U/g).
  • Characteristic chromatogram (8 common peaks detected by HPLC).
  • Safety (aflatoxin B1 < 2μg/kg).

Industrial Achievements

A leading pharmaceutical enterprise adopted this ，reducing batch-to-batch variation coefficient from 28% to 4.3% and improving clinical efficacy by 19%. The technology has been granted a national invention patent (ZL202010345678.9), setting a benchmark for standardized fermentation of traditional TCMs.

Case 2: Monascus – High-Efficiency Preparation Driven by Metabolic Engineering

Monascus, valued for its bioactive compounds like Monacolin K (a cholesterol-lowering agent), has undergone transformative improvements through genetic engineering и precision fermentation control.

Strain Modification Strategy

Using the CRISPR-Cas9 system, the citrinin synthesis gene cluster (cts gene) in Monascus purpureus M-7 was knocked out to eliminate toxic byproducts, while key enzymes for lovastatin synthesis (lovC, lovD) were overexpressed. This modification boosted Monacolin K yield to over 12g/L, a breakthrough in industrial production.

Fermentation Process Optimization

• Innovative Culture Medium: Agricultural waste (rice husk powder: wheat bran = 3:1) is used as the matrix, with 0.2% tea polyphenols added to induce secondary metabolism, reducing production costs and enhancing sustainability.
• Precise Parameter Control:
  • Dissolved oxygen strategy: 40% in 0-48h; 25% in 49-144h (tailored to different growth stages).
  • pH regulation: maintained at 5.5 for the first 48h, then allowed to decrease naturally.
  • Feeding strategy: 0.5% glycerol + 0.1% magnesium sulfate added at 48h to sustain metabolite synthesis.
• Real-Time Monitoring: Integration of near-infrared spectroscopy (NIR) and electronic nose technology enables real-time tracking of biomass and metabolite changes, ensuring process stability.

Quality Control Breakthrough

A “dual-standard, dual-detection” mechanism is implemented:

• Lovastatin content (HPLC, ≥1.5%).
• Citrinin limit (LC-MS/MS, <0.2ppm).
• Fungal toxin screening (untargeted analysis via UPLC-Q-TOF/MS), ensuring safety and efficacy.

Case 3: Astragalus – Efficacy Enhancement via Probiotic-Directed Transformation

Astragalus, a staple in TCM for immune modulation, has seen enhanced bioactivity through probiotic co-fermentation, leveraging microbial metabolism to unlock hidden therapeutic potential.

Strain Screening System

Lactobacillus plantarum LP-3 (oxygen- and TCM-tolerant) and Bacillus subtilis B-12 (cellulase-producing) were isolated from traditionally fermented astragalus. Their co-fermentation conditions were optimized using response surface methodology to maximize synergistic effects.

Technological Innovations

• Pretreatment Technology: Steam explosion-enzymolysis combined treatment (1.5MPa for 5min, followed by 0.5% cellulase at 50°C for 2h) increased astragalus polysaccharide dissolution rate from 18% to 42%, facilitating microbial access to nutrients.
• Fermentation Control:
  • Inoculation timing: B-12 first, followed by LP-3 after 24h.
  • Temperature regulation: 37°C for the first 48h, then reduced to 30°C to promote secondary metabolites.
  • Humidity management: Dynamic humidification to maintain 45%-50% moisture content.
• Metabolite Analysis: Newly formed astragaloside IV derivatives (molecular weight reduced to 789.4Da) were identified, with isoflavone aglycone content increasing 3.2-fold, enhancing bioavailability.

Efficacy Validation

• Immunomodulation: Fermented astragalus increased mouse spleen index by 41%, significantly higher than the unfermented group (P < 0.01).
• Antioxidant capacity: DPPH free radical scavenging rate rose from 28% to 67%.
• Gut microbiota regulation: Firmicutes/Bacteroidetes ratio decreased from 1.2 to 0.7, improving intestinal barrier function.

Industrial Applications of Fermented TCM

Fermented TCM has expanded beyond pharmaceuticals into agriculture and animal husbandry, demonstrating versatile value:

1. Enzyme-Microbe Synergistic Fermentation in Livestock Breeding
   • Formula: Astragalus, Codonopsis, Atractylodes macrocephala combined with cellulase and protease.
   • Technological edge: Lactic acid bacteria create an acidic environment ideal for enzymatic reactions; compound enzymes target plant cell walls, increasing active ingredient dissolution by 3-5 times.
   • Effects: Fermented astragalus polysaccharide absorption rate improved 4-28 times; sow constipation rate reduced by 60%, piglet survival rate increased by 15%; broiler feed intake rose by 12%, feed conversion ratio decreased by 8%.
2. Fermented Formulation for Hypertension Treatment
   • Patent technology uses probiotic fermentation to prepare anti-hypertensive TCM:
     • Ingredients: Atalantia buxifolia root (30g/L), milk thistle fruit (30g/L), Erythropalum scandens (10g/L), wampee dried fruit (10g/L), Ficus deltoidea (5g/L).
     • Process: Ultra-fine grinding for cell wall breaking; sterilization at 110°C for 30min; inoculation with 5% lactic acid bacteria (3% Lactobacillus rhamnosus + 2% Lactobacillus plantarum); anaerobic fermentation at 37°C for 48h.
     • Product form: Fermented liquid (direct filling) or spray-dried powder.
3. Fermented TCM Formula for Layer Chicken Breeding
   • Patent formula by Shanxi Dayu Bioengineering Co., Ltd.: A combination of Acanthopanax senticosus, motherwort, astragalus, and Artemisia annua.
   • Process features: Bacillus coagulans fermentation broth preparation; solid-state fermentation; fluidized bed drying.
   • Effects: Layer egg production rate increased by 4.5%, feed-egg ratio decreased by 6%.

Future Trends in Fermented TCM

Research into fermented TCM is rapidly advancing, with key focuses on:

• Strain Breeding: Directional cultivation of engineered strains with specialized tolerance to TCM components and enhanced metabolic capabilities.
• Fermentation Equipment Innovation: Devices like the rotary spray fermentation tank (developed by Anhui Tiankang Group) enable comprehensive cleaning and sterilization, reducing contamination risks.
• Multi-Strain Synergy: Exploring staged multi-strain fermentation to address complex TCM component transformation.
• Multi-Omics Applications: Metatranscriptomics to analyze enzyme expression dynamics during Liushenqu fermentation, identifying 12 key glycoside hydrolase genes, paving the way for precise metabolic regulation.

Challenges remain, such as defining Q-Markers (quality markers) by integrating pharmacodynamics and metabolomics data, and aligning with ICH Q13 guidelines for international registration. However, with the 5th Academic Conference of the World Federation of Chinese Medicine Societies’ TCM Fermentation Research Committee (late 2024), the field is poised for accelerated growth.

Fermented TCM, bridging ancient wisdom and modern science, is redefining TCM’s potential. Through standardized processes, genetic engineering, and innovative applications, it is set to become a cornerstone of global natural medicine, offering safer, more effective, and scalable solutions for health and agriculture.