The application of lecithin mixed reverse micelles for bioproduction and separation processes

卵磷脂混合反胶束在生物生产和分离过程中的应用

• 批准号: 06650914
• 负责人: KUBOI Ryoichi
• 金额: $ 1.28万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06650914/
• 关键词: Riverse micelle; Mixed micells; Lecithin; AOT; Partition chromatograph; CPC; Bioreactor; beta-Galactosidase; 遠心溶液分配クロマト

The application of reverse micellar micro bioreactor systems for enzymatic conversion with continuous recovery of water-soluble products was demonstrated, based on the production of galactose by micelles containing beta-galactosidase. The beta-gal was firmly encapsulated in AOT (bis (2-ethylhexyl) sodium sulfosuccinate) /lecithin mixed micelles. The addition of lecithin to the AOT reverse micelles stabilized the micellar interface, and the leakage of the encapsulated beta-gal to the bulk aqueous phase was therefore negligible, while substrate and product were free to transfer and partition owing to their low molecular weights. The stability of the solubilized beta-gal was also increased in the AOT/lecithin mixed micelles as compared to the conventional AOT micelles. A consider ably higher activity was observed in the present systems than in the bulk buffer solution. Enzymatic conversion and product recovery was carried out in a continuous operation mode by use of centrifugal partition chromatography (CPC), employing the reverse micellar solution with encapsulated beta-gal as the stationary phase and the substrate aqueous solution as the mobile phase. Use of the CPC contactor eliminated any mass transfer limitation on the substrate and the product, and galactose was continuously recovered in the effluent.

以含有β -半乳糖苷酶的胶束生产半乳糖为基础，展示了反胶束微生物反应器系统在酶促转化中连续回收水溶性产物的应用。β -gal被牢固地包裹在AOT（双（2-乙基己基）琥珀酸钠）/卵磷脂混合胶束中。在AOT反胶束中加入卵磷脂稳定了胶束界面，因此被封装的β -gal向体水相的泄漏可以忽略不计，而底物和产物由于其低分子量而可以自由转移和分裂。与传统的AOT胶束相比，AOT/卵磷脂混合胶束中溶解的β -gal的稳定性也有所提高。在本系统中观察到的活性比在散装缓冲液中观察到的活性高得多。采用离心分配色谱法（CPC），以包封β -半乳糖的反胶束溶液为固定相，底物水溶液为流动相，在连续操作模式下进行酶转化和产物回收。CPC接触器的使用消除了对底物和产物的传质限制，并且在出水中连续回收了半乳糖。

项目成果

K.Shiomori, R.Kuboi, I.Komasawa and S.Kawano: "Effective Purification Method of Large Molecular Weight Proteins Using Conventional AOT Reverse Micelles" J.Chem.Eng.Japan. 28. 803-809 (1995)

K.Shiomori、R.Kuboi、I.Komasawa 和 S.Kawano：“使用传统 AOT 反胶束的大分子量蛋白质的有效纯化方法”J.Chem.Eng.Japan。

Y.Yamada: "Effective Bioconuersion with Continuous Product Recovery Using AOT/Lecithin Mixed Reverse Micollar Systems and Centritugal Portition Chromatography as a Novel Bioreactor" Biotechnol.Prog.11. 682-688 (1995)

Y.Yamada：“使用 AOT/卵磷脂混合反向微领系统和离心部分色谱作为新型生物反应器进行连续产品回收的有效生物转化”Biotechnol.Prog.11。

K.Shiomori: "Effective Puritication Method of Large Molecular Weight Proteins Using Conventional AOT Reverse Micelles" J.Chem.Eng.Japan. 28. 803-809 (1995)

K.Shiomori：“使用传统 AOT 反胶束的大分子量蛋白质的有效纯化方法”J.Chem.Eng.Japan。

Y.Yamada, R.Kuboi and I.Komasawa: "Effective Bioconversion with Continuous Product recovery Using AOT/Lecithin Mixed Reverse Micellar Systems and Centrifugal Partition Chromatography as a Novel bioreactor" Biotechnol.Prog.11. 682-688 (1995)

Y.Yamada、R.Kuboi 和 I.Komasawa：“使用 AOT/卵磷脂混合反胶束系统和离心分配色谱作为新型生物反应器进行有效的生物转化和连续产品回收”Biotechnol.Prog.11。

Y.Yamada,: "Effective Bioconversion with Continuous Product Recovery Using AOT/Lecithin Mixed Reverse Micellar Systems and Centrifugal Partition Chromatography as a Novel Bioreactor" Biotechnol.Prog.,. 11. 682-688 (1995)

Y.Yamada，：“使用 AOT/卵磷脂混合反胶束系统和离心分配色谱作为新型生物反应器进行连续产物回收的有效生物转化”Biotechnol.Prog.,。