Identification of Behavior of Protein in Various Environments and Its Application to Downstream Processing
Grant-in-Aid for Scientific Research (C).
|Research Institution||Kanai University|
YAGI Hideharu Kansai University, Faculty of Engineering, Professor, 工学部, 教授 (70109891)
|Project Fiscal Year
1997 – 1999
Completed(Fiscal Year 1999)
|Budget Amount *help
¥3,400,000 (Direct Cost : ¥3,400,000)
Fiscal Year 1999 : ¥500,000 (Direct Cost : ¥500,000)
Fiscal Year 1998 : ¥1,300,000 (Direct Cost : ¥1,300,000)
Fiscal Year 1997 : ¥1,600,000 (Direct Cost : ¥1,600,000)
|Keywords||Protein / Refolding / Hydrophobic interaction / Aggregation / Crystallization / Biopolymer / Dynamic membrane / タンパク質 / リフォルディング / 疎水性相互作用 / 凝集 / 晶折 / バイオポリマー / ダイナミック膜 / クロマトグラフィー / 精密ろ過 / 分離精製|
The separation of the target material from the impurities requires the recognition of difference in the behavior or morphology in the solution in addition to the physical properties. The behavior and morphology affect the biological activity of the protein or its components. The aggregation of protein molecules results in failure of refolding of denatured protein but forms the dynamic membrane which is expected as a recyclable membrane for ultrafiltration. The aggregation of molecules in the solution is also the first step of crystallization.
This research project deals with the following themes from the standpoint of identification of behavior of protein in various environments and its application to downstream processings :
1. Application of hydrophobic interaction to the separation of protein during refolding.
The adsorption characteristics were different between the native and denatured lysozyme. Reactivated lysozyme by semibatch refolding was recovered by the column chromatography el
2. Refolding of denatured enzymes in semibatch system
The denatured protein solution was supplied into the renaturation solution at a constant flow rate. The recoveries of activity in a semibatch operation were higher than those in batch operations.
3. Characteristics of formation of dynamic membrane on porous ceramic support.
The dynamic membrane of ovalbumin with self-rejection over 0.9 was formed on the porous ceramic support. The addition of salt or inhibitor of gelation inhibited the formation of membrane and lost the already formed membrane.
4. Effect of coexinsting amino acids or impurity on the crystallization of L-aspartic acid.
Coexisting amino acids were included fairly in the product L-aspartic crystals and detoriarated the crystal habit.
5. Separation by two-step filtration of biopolymer formed by Bacillus subtilis.
The biopolymer, γ-PGA, formed in Bacillus subtilis cultivation was separated from the microorganism by microfiltration and recovered by subsequent ultrafiltration. The fouling for the microfiltration was improved by back flushing with the permeate. Less
Research Output (8results)