Medicine composition-compound digestive enzyme capsule (II) and preparation method thereof
The technology of compound digestive enzyme capsule and capsule shell is applied in the field of biomedicine, can solve the problems of affecting the use efficiency of pepsin, the pepsin cannot be released quickly, is damaged, etc., and achieves convenient administration, storage and transportation, saving preparation cost, and simplifying The effect of craftsmanship
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Embodiment 1
[0030] Example 1 Preparation of trypsin
[0031] We have designed the optimal expression sequence of the trypsin gene of the present invention in yeast, its nucleotide is shown in SEQ ID NO: 1, and the amino acid sequence of the trypsin encoded by it is shown in SEQ ID NO: 2. We commissioned Shanghai Shenggong Company to artificially synthesize the gene sequence and clone it into yeast cells according to conventional methods to construct expression engineering bacteria. In short, using the synthesized gene sequence as a template, using the The upstream and downstream primers were amplified by PCR, and then connected between EcoR I and Xba I of pPICZαA (available from Invitrogen, USA), and the positive construct was electrotransformed into yeast GS115 strain (available from Invitrogen, USA), and screened with Zeocin The positive yeast cells were used as engineering bacteria.
[0032] Inoculate the constructed engineered bacteria into 25ml seed culture medium (formulation: 2% ...
Embodiment 2
[0033] Example 2 Study on properties of trypsin
[0034] The trypsin prepared in Example 1 was dissolved in pH 8.0 Tris-HCl buffer solution containing 20 mM calcium chloride, put into a dialysis bag, and dialyzed twice in pH 8.0 Tris-HCl buffer solution (0.1M) at 4°C. Each time, 15 minutes each time, then adding Tos-Gly-Pro-Arg-4-pNA (Chromozym TRY) as a substrate, autoactivating trypsin at different temperatures for 1 hour, and then detecting its activity. The results are shown in Table 1. The trypsin of the present invention has high self-activation at physiological temperature, but almost no activity at low temperature, and the enzyme activity at 37°C can reach more than 300U / mg in a short time.
[0035] Table 1 Properties of pancreatin of the present invention
[0036] temperature (°C) 4 25 37 50 Activity (U / mg) 2.8 276 311 65
Embodiment 3
[0037] The preparation of embodiment 3 trypsin enteric-coated granules
[0038] (1) Dissolve hydroxypropylmethylcellulose in water, then add 95% ethanol solution to prepare 2% (W / W) hydroxypropylmethylcellulose in ethanol aqueous solution (ethanol: water (volume ratio) = 1: 1).
[0039] (2) Mix 28 parts of pancreatin, 8 parts of sucrose and 4 parts of dextrin prepared in Example 1 and put them in a centrifugal granulator (available from Shandong Linyi Huanshi Micropill Equipment Co., Ltd., BZL-300 type) According to the manufacturer's instructions, spray 32 parts of the ethanol aqueous solution of hydroxypropyl methylcellulose in step (1) (it has been measured that about 0.6 parts of hydroxypropylmethylcellulose is bound to the core of the pellet), and granulate to form The ball core, after drying, has an average particle size of 0.9mm and is ready for use.
[0040] (3) Add 3 parts of triethyl citrate to 500 parts of distilled water, mix well and then transfer into ...
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