ABSTRACT
Objective:To optimize the extraction process of the water extract of Zhenwutang and study on the preparation of gran -ule of Zhenwu decoction to provide reference for the development and utilization of Zhenwutang granule .Methods: Heating refluxing was used, and the effects of the ratio of solid to liquid , extraction time and times were investigated by orthogonal test .As the synthetic indices of evaluation , the yield of extraction and the contents of paeoniflorin and benzoylmesaconine measured by HPLC were deter -mined to confirm the optimal water extraction process of Zhenwutang granule .Besides, granularity pass rate, moisture, loss on drying, solubility and angle of repose of the granule were regarded as the indices to evaluate the best ratio of the excipients in the preparation of the granule by single factor test.Results:Paeoniflorin and benzoyl mesaconitine had good linearity within the range of 5.45-32.70μg (r=0.9996) and 3.24-16.80 μg(r=0.9997), respectively.The average recovery was 99.62％ (RSD =1.34％ , n=6) and 1017.2 ％(RSD=1.74％, n=6), respectively.The optimum extraction process was as follows :the ratio of solid to liquid was 1:12 with twice refluxing extraction ( 2h for each time) .The optimum granule forming process was as follows:the pharmaceutical excipients were a mixture of dextrin and soluble starch with the best ratio of 1:3. The granularity pass rate , moisture, loss on drying, solubility and angle of repose of the granule was94 .12％ ,4.87 ％, 0.93％,89 .23％ and 36.18°, respectively.Conclusion:The optimized re-fluxing extraction process is stable , reliable and feasible , and the prepared granule is in good formability and melting .
ABSTRACT
In this study, the mature peptide sequence of a pectin lyase gene A was amplified from Aspergillus niger strain EIM-6 by using RT-PCR reverse transcription technique. The cloned gene was then inserted into a Pichia pastoris expression vector pPIC9k to produce the recombinant expression plasmid pPIC9K-pelA. By using electric shocks, we successfully transformed the recombinant pPIC9K-pelA into Pichia pastoris GS115. The activity of the engineered strain reached to 2.3 U/mL after induction with the final concentration of 1.5% methanol. SDS-PAGE analysis revealed that the pPIC9K-pelA transformant had an additional protein band of approximately 38 kD, which was not present in the control. There were no significant differences between the recombinant and native pectin lyase with regard to their hydrolysis activities.