Shandong University of Traditional Chinese Medicine Announces Transfer of Bile-Tolerant Bovine Calculus-Transforming Bacterial Technology Package for RMB 3 Million
Recently, Shandong University of Traditional Chinese Medicine released a public notice on the transformation of scientific and technological achievements, proposing to transfer its research outcomes through assignment.“A Bile-Tolerant Bezoar-Transforming Bacterial Strain and Its Application” and “A Screening Medium for Bile-Tolerant Bezoar-Transforming Bacteria, Its Preparation Method and Application”Transfer of two patent portfolios, with a minimum transfer price of3 million yuan. The inventor of this patent isWang Houwei and His Team。
Image from the official website of the Shandong Provincial Technology Property Rights Trading Platform
The core of this technology package isScreening yielded a novel bovine bile-tolerant strain for bezoar transformation—Citrobacter bovis NDZNM-01, and established a fermentation production process for applying this strain in the in vitro cultivation of Calculus Bovis. The essence is to address industry pain points such as the scarcity of natural Calculus Bovis, high production costs, and low conversion efficiency associated with existing in vitro cultivated Calculus Bovis, thereby achieving efficient, low-cost production of in vitro cultivated Calculus Bovis.
As a precious traditional Chinese medicinal material, bezoar (Calculus Bovis) is in high demand for clinical applications and the production of proprietary Chinese medicines; however, natural bezoar, due toHigh acquisition difficulty, scarce production, and persistent supply-demand imbalance, has become the core bottleneck in the development of the traditional Chinese medicine (TCM) industry. The synthetic cow-bezoar developed in the 1950s was a formulated preparation with significant differences in composition and efficacy compared to natural cow-bezoar, making it unsuitable as a substitute in most scenarios. Although in vitro cultured cow-bezoar, successfully developed in the 1990s, closely resembles natural cow-bezoar in chemical composition and clinical efficacy, and is included in the Pharmacopoeia as an equivalent medicinal ingredient, its widespread and affordable application has been hindered by production technology bottlenecks, severely restricting industry growth.
From an industry perspective, the existing production model for in vitro cultured Calculus Bovis has significant shortcomings, with the core issue lying inDual Deficiencies in Strain Performance and Conversion Efficiency。
On the one hand, bovine bile exhibits broad-spectrum bacteriostatic activity,Current production strains struggle to survive long-term in bovine bile., most highly efficient bacteria for the biotransformation of Calculus Bovis survive in bovine bile for no more than 48 hours, failing to establish a stable biotransformation system. This directly results in low conversion efficiency of the core components of Calculus Bovis, making it difficult to meet the quality specifications stipulated in the Pharmacopoeia. To achieve the required content standards, existing manufacturers must add substantial amounts of composite calcium bilirubinate and bile acids during the cultivation process. This not only significantly increases raw material and production process costs but also drives up the market price of the final product, with its selling price exceeding one-third that of natural Calculus Bovis, thereby hindering clinical application and market adoption.
On the other hand,Current production strains for in vitro cultured Calculus Bovis lack specificity and standardization., no Citrobacter strains capable of tolerating ox bile have been identified to date. The lack of standardized protocols for strain identification and cultivation results in poor process stability and significant batch-to-batch variability, further increasing production control costs and quality risks. Meanwhile, existing processes fail to optimize fermentation conditions based on strain-specific characteristics, lacking precise control standards for parameters such as pH and temperature. This prevents the maximization of the strains' biotransformation capacity, creating a vicious cycle of "low strain efficiency → addition of excipients → high costs → expensive pricing."
Furthermore,Production Rights for In Vitro Cultured Calculus Bovis Are Highly Concentrated, with only a single enterprise achieving exclusive production, the industry lacks technological competition and iterative innovation, making it difficult to upgrade production processes and reduce costs, thereby failing to meet the demand of the traditional Chinese medicine industry for large-scale, low-cost raw bezoar.
These issues have directly resulted in a supply dilemma in the raw material market for Calculus Bovis:Natural Calculus Bovis is scarce and difficult to obtain, synthetic Calculus Bovis has insufficient efficacy, and in vitro cultured Calculus Bovis is prohibitively expensive with limited supply. Downstream sectors, including the production of proprietary Chinese medicines and clinical diagnosis and treatment, are all constrained by raw material shortages, creating a bottleneck for the large-scale development of the traditional Chinese medicine industry.
For the raw material industry of Calculus Bovis, existing production technologies can no longer meet the core clinical and market demands for high-quality, low-cost, large-scale Calculus Bovis. There is an urgent need for a dedicated bile-tolerant, high-conversion bacterial strain for Calculus Bovis transformation, coupled with standardized fermentation processes, to break through the industrial bottlenecks in Calculus Bovis production.
This technology, involving a bile-tolerant bacterial strain for the transformation of Calculus Bovis and its application in the in vitro cultivation of Calculus Bovis, addresses existing challenges in current Calculus Bovis production through the screening of novel proprietary strains and the establishment of standardized fermentation processes.Poor strain tolerance, low conversion efficiency, and high costsaddressing the industry’s pain points, it delivers disruptive advantages in strain performance, conversion efficiency, and production compatibility, providing a novel solution for the large-scale, low-cost production of natural bezoar raw materials.
FromCore Strain InnovationFrom the perspective of, patents are based onBile Tolerance and High-Efficiency Transformation Capability, to establish a high-quality strain foundation for the in vitro cultivation of cattle gallstones.
On the one hand,The screened Citrobacter bovis strain NDZNM-01 is the first identified bile-resistant Citrobacter bovis., breaking through the technical barrier that previously prevented Citrobacter from surviving in bovine bile. It survives for more than 9 days in pure bovine bile, far exceeding the 48-hour survival limit of existing strains. The strain exhibits a lag phase within the first 48 hours, with bacterial density reaching its peak at 96 hours. It is capable of forming a stable biotransformation system in bovine bile, which possesses broad-spectrum bacteriostatic properties, thereby fundamentally addressing the core issues of short strain viability and instability of the transformation system; on the other hand,The transformation efficiency of this strain has achieved a qualitative leap., fermented at 30°C for 16 days, the bilirubin content in bovine bile reached 2.01‰ and the cholic acid content reached 6.42%, representing a 2.35-fold and 2.13-fold increase, respectively, compared to natural bovine bile. This process meets the pharmacopoeial quality specifications without the need for additional composite calcium bilirubinate or bile acids, thereby completely breaking the vicious cycle of “low-efficiency strains–excipient addition–high costs” prevalent in existing production methods.
Furthermore, the patent achieves dual standardization in strain identification and production processes, balancing strain uniqueness with production compatibility.
In terms of strain identification, a proprietary molecular identification system has been established., which clarifies the characteristic 16S rDNA base sequence of the strain, six characteristic base sites, and specific forward (F) and reverse (R) amplification primers. This enables rapid and accurate identification of the strain through PCR amplification combined with base site comparison, ensuring the uniqueness and purity of the strain. It addresses the issues of lacking unified identification standards and significant batch-to-batch variability in existing strains. Regarding culture medium preparation, the scale-up cultivation of the strain uses tryptone, beef extract, and sodium chloride as core raw materials. Agar powder can be added as needed to prepare solid or semi-solid media. The raw materials are readily available, and the formulation is simple, significantly reducing the material and process costs for strain cultivation.
In terms of fermentation process, precise standardized control parameters have been established., with the fermentation broth pH clearly controlled at 6.5–7.5 (optimal: 7.2) and the fermentation temperature at 20–35°C (optimal: 30°C). The influence of pH and temperature on conversion efficiency was experimentally verified, enabling precise regulation according to production needs to maximize the strain’s conversion capacity; meanwhile, this processAdapted for Industrial-Scale Production, with clear strain growth curves and controllable fermentation cycles, enabling stable batch production, thereby addressing the pain points of non-standardized production processes and unstable product quality in existing in vitro cultured Calculus Bovis manufacturing.
These advantages directly empower the development of the bezoar industry: bile-resistant, high-conversion bacterial strains enhance production efficiency at its source, significantly reducing raw material and excipient costs, thereby bringing the price of in vitro cultivated bezoar back to a reasonable range; standardized strain identification and fermentation processes ensure stability in production and uniformity in product quality, laying the foundation for large-scale industrial manufacturing; the discovery of a novel Citrobacter species from bovine bile fills the application gap of Citrobacter in bezoar biotransformation, providing new directions for the research and development of bezoar-converting microbial strains.
For the raw material industry of Calculus Bovis, this patented technology not only breaks through the existing technical bottlenecks in production but also achieves, through strain innovation and process standardization,Bezoar Bovis: Production Efficiency, Product Quality, and Cost ControlThe triple enhancement has made the widespread accessibility of precious traditional Chinese medicine bezoar possible, providing core raw material support for the large-scale development of the traditional Chinese medicine industry.
Currently, in response to the industry pain points of a significant supply-demand gap in the raw Calculus Bovis market, low production efficiency, and high costs, domestic pharmaceutical companies and research institutions are accelerating R&D and industrial layout related to Calculus Bovis cultivation. This has formed a competitive market landscape centered on upgrading in vitro cultivation processes, screening bile-tolerant bacterial strains for transformation, and supplementing with imported natural Calculus Bovis, while artificially synthesized Calculus Bovis serves as a basic raw material to meet lower-end market demands.
Wuhan Jianmin Dapeng Pharmaceutical Co., Ltd. is the sole domestic manufacturer of in vitro cultivated Calculus Bovis., its core productsIn Vitro Cultured Calculus BovisListed in the Chinese Pharmacopoeia (2020 Edition), it can be used as a medicinal substitute for natural Calculus Bovis. In April 2025, the product completed the public notice period for the initial acceptance of its application for Traditional Chinese Medicine Variety Protection by the National Medical Products Administration. Leveraging the core technology for in vitro cultured Calculus Bovis developed by Professor Cai Hongjiao, the enterprise simulates the gallstone formation process through co-culturing bovine bile with microorganisms.
In the field of natural Calculus Bovis, China is expanding supply channels through import pilot policies to alleviate raw material shortages. In January 2026,Zhangzhou Pientzehuang Pharmaceutical Co., Ltd.Completed China’s first import of natural bovine bezoar from Argentina, becoming the first practical case following the implementation of the pilot policy on natural bovine bezoar imports by the National Medical Products Administration and the General Administration of Customs; in March 2026,Beijing TongrentangSecured China’s first customs clearance certificate for bezoar pharmaceuticals, completing the first import transaction of natural bezoar in over two decades. All imported bezoar originates from BSE-free zones certified by the World Organisation for Animal Health (WOAH), ensuring safe and controllable raw material sourcing. Meanwhile, Announcement No. 25 of 2026 issued by the General Administration of Customs explicitly permits the export of Uruguayan bezoar to China starting in March, further diversifying the supply sources for natural bezoar imports.
The Citrobacter freundii strain NDZNM-01, selected through this technology, overcomes the traditional bottlenecks of poor bile tolerance and low conversion efficiency. The accompanying standardized fermentation process significantly reduces the production cost of in vitro cultured Calculus Bovis, fundamentally addressing the industry’s pain points of supply-demand imbalance and high prices for raw Calculus Bovis.
This technology fills the application gap of Citrobacter in the field of Bezoar Bovis (Calculus Bovis) transformation, laying a core technological foundation for the industrialized and large-scale production of in vitro cultured Bezoar Bovis. With the future industrial implementation of this bacterial strain and its associated processes, it will promote the widespread availability and affordability of raw Bezoar Bovis in the market. Furthermore, it provides new insights and directions for the research and development of biosynthetic technologies for precious traditional Chinese medicine (TCM) ingredients, thereby facilitating the high-quality upgrading of the TCM industry.