Scale-up of fermentation and purification of recombinant allophycocyanin over-expressed in Escherichia coli

doi:10.1016/j.procbio.2005.02.023

IOCAS-IR > 实验海洋生物学重点实验室

	Scale-up of fermentation and purification of recombinant allophycocyanin over-expressed in Escherichia coli
	Ge, BS ; Tang, ZH ; Zhao, FQ ; Ren, YH ; Yang, Y; Qin, S
	2005-10-01
发表期刊	PROCESS BIOCHEMISTRY
ISSN	1359-5113
卷号	40 期号:10 页码:3190-3195
文章类型	Article
摘要	Phycobiliprotein is a photosynthetic antenna pigment found in cyanobacteria, rhodophytes, cryptophytes and certain dinoflagellates, which has been found to have anti-oxidative and anti-tumour activities. In this paper, a recombinant allophycocyanin (rAPC) had been expressed in Escherichia coli for anti-tumour effect. E. coli cells were cultured using glucose fed-batch method to achieve high cell densities. The biomass of rAPC was up to 3.52 g/L broth. The rAPC was purified from soluble E. coli cell lysate employing hydrophobic interaction chromatographic (HIC) method developed at the bench scale using 20 mL column. The process was performed at the pilot scale using 500 mL column for evaluation of scale-up. An amylose affinity column was used to improve the purity of final product in pilot scale purification. The purification process resulted in greater than 98% pure product and yielded up to 2.0 g/kg wet cells at the bench scale and 1.2 g/kg wet cells at the pilot scale. Peptide mapping was used to prove the identity of rAPC purified from bench scale and pilot scale process. Purified rAPC at the pilot scale was found to have remarkable inhibition on S-180 carcinoma in mice. (c) 2005 Elsevier Ltd. All rights reserved.; Phycobiliprotein is a photosynthetic antenna pigment found in cyanobacteria, rhodophytes, cryptophytes and certain dinoflagellates, which has been found to have anti-oxidative and anti-tumour activities. In this paper, a recombinant allophycocyanin (rAPC) had been expressed in Escherichia coli for anti-tumour effect. E. coli cells were cultured using glucose fed-batch method to achieve high cell densities. The biomass of rAPC was up to 3.52 g/L broth. The rAPC was purified from soluble E. coli cell lysate employing hydrophobic interaction chromatographic (HIC) method developed at the bench scale using 20 mL column. The process was performed at the pilot scale using 500 mL column for evaluation of scale-up. An amylose affinity column was used to improve the purity of final product in pilot scale purification. The purification process resulted in greater than 98% pure product and yielded up to 2.0 g/kg wet cells at the bench scale and 1.2 g/kg wet cells at the pilot scale. Peptide mapping was used to prove the identity of rAPC purified from bench scale and pilot scale process. Purified rAPC at the pilot scale was found to have remarkable inhibition on S-180 carcinoma in mice. (c) 2005 Elsevier Ltd. All rights reserved.
关键词	Scale-up Recombinant Allophycocyanin Escherichia Coli Protein Purification Tumour Inhibition
学科领域	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical
DOI	10.1016/j.procbio.2005.02.023
URL	查看原文
收录类别	SCI
语种	英语
WOS记录号	WOS:000231833100006
引用统计	被引频次：20[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/3094
专题	实验海洋生物学重点实验室
作者单位	1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Ge, BS,Tang, ZH,Zhao, FQ,et al. Scale-up of fermentation and purification of recombinant allophycocyanin over-expressed in Escherichia coli[J]. PROCESS BIOCHEMISTRY,2005,40(10):3190-3195.
APA	Ge, BS,Tang, ZH,Zhao, FQ,Ren, YH,Yang, Y,&Qin, S.(2005).Scale-up of fermentation and purification of recombinant allophycocyanin over-expressed in Escherichia coli.PROCESS BIOCHEMISTRY,40(10),3190-3195.
MLA	Ge, BS,et al."Scale-up of fermentation and purification of recombinant allophycocyanin over-expressed in Escherichia coli".PROCESS BIOCHEMISTRY 40.10(2005):3190-3195.

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