Investigation of chromatography and polymer/salt aqueous two-phase processes for downstream processing development of recombinant phenylalanine dehydrogenase

被引：13

作者：

Omidinia, Eskandar ^{[1
]}

Mohamadi, Hamid Shahbaz ^{[1
]}

Dinarvand, Rassoul ^{[2
]}

Taherkhani, Heshmat-Allah ^{[3
]}

机构：

[1] Pasteur Inst Iran, Dept Biochem, Tehran 13164, Iran

[2] Univ Tehran Med Sci, Fac Pharm, Tehran, Iran

[3] Golestan Univ Med Sci, Fac Med, Gorgan, Iran

来源：

BIOPROCESS AND BIOSYSTEMS ENGINEERING | 2010年 / 33卷 / 03期

关键词：

Aqueous two-phase systems (ATPS); Chromatography; Downstream processing; Phenylalanine dehydrogenase (PheDH); PURIFICATION; EXTRACTION; SYSTEMS; RECOVERY;

D O I：

10.1007/s00449-009-0327-8

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

This work presents a comprehensive study between the polymer/salt aqueous two-phase systems (ATPS) and chromatography process for downstream processing of recombinant Bacillus badius phenylalanine dehydrogenase (PheDH). First, the partitioning behavior of recombinant PheDH in polyethylene glycol (PEG)/K2HPO4 ATPS was examined. For comparative purpose, a classical chromatographic protocol was performed as well. Investigation of chromatography and ATPS procedures revealed that the ATPS comprising of 9% (w/w) PEG-6000, 16% (w/w) K2HPO4 and 16% (w/w) KCl with pH of 8.0, volume ratio (V (R) ) of 0.25, temperature of 25 A degrees C and 40% (w/w) cell lysate ensured the most favorable approach for PheDH downstream process. A specific activity of 4,231.4 U/mg, a yield of 96.7% and a recovery of 162.0% were obtained. Furthermore, the shorter process time (4 vs. 48 h) and the lower total cost (4 vs. 20 a,not sign) were additionally features that confirmed the suitability of proposed technique.

引用

页码：317 / 329

页数：13

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