The invention discloses an efficient and stable in-vitro cultured
calculus bovis preparation process, and belongs to the technical field of biological preparation of traditional Chinese medicines. The process comprises the five steps of
genetic engineering compound
enzyme liquid preparation, bile pretreatment, bionic enzymatic reaction,
crystallization purification and quality regulation and control. The core innovation points are as follows: an
engineering strain is constructed through a
CRISPR-Cas9 technology to obtain a high-activity compound
enzyme solution (the activity is greater than or equal to 80U / mL); an AI
intelligent control system is adopted to realize accurate regulation and control, the stability fluctuation of the
system is less than or equal to + / -0.5%, and the
contamination rate is less than or equal to 0.03%; a composite anti-oxidation
system and a supercritical COrefining process are matched, so that the oxidation loss of effective components is reduced. The production period is shortened to 3 days, the finished product
bilirubin is larger than or equal to 38.2%, the
cholic acid is larger than or equal to 6.6%, the product conforms to the 2025 edition
pharmacopoeia standard,
acute toxicity is extremely low,
skin irritation and
sensitization are avoided, the industrialization prospect is good, the product quality is high, safety is high, and natural
calculus bovis can be effectively replaced to meet clinical requirements.