Cost-effective production of recombinant peptides in Escherichia coli

被引:55
|
作者
Gaglione, Rosa [1 ]
Pane, Katia [2 ]
Dell'Olmo, Eliana [1 ]
Cafaro, Valeria [3 ]
Pizzo, Elio [3 ]
Olivieri, Giuseppe [4 ]
Notomista, Eugenio [3 ]
Arciello, Angela [1 ,5 ]
机构
[1] Univ Naples Federico II, Dept Chem Sci, I-80126 Naples, Italy
[2] IRCCS, SDN, I-80143 Naples, Italy
[3] Univ Naples Federico II, Dept Biol, I-80126 Naples, Italy
[4] Wageningen Univ & Res, AlgaePARC, Bioproc Engn, POB 16, NL-6700 AA Wageningen, Netherlands
[5] INBB, Rome, Italy
关键词
Host defence peptides; Apolipoprotein B; Apolipoprotein A-I; Auto-inducing expression procedure; Cost-effective recombinant production; Techno-economic analysis; HOST-DEFENSE PEPTIDES; ANTIMICROBIAL PEPTIDES; EFFECTIVE EXPRESSION; APOA-I; AUTOINDUCTION; LACTOSE; PURIFICATION; DOMAIN; FORMS;
D O I
10.1016/j.nbt.2019.02.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Among bioactive peptides, cationic antimicrobial peptides (AMPs), also referred to as host defence peptides (HDPs), are valuable tools to treat infections, being able to kill a wide variety of microbes directly and/ or modulate host immunity. HDPs have great therapeutic potential against antibiotic-resistant bacteria, viruses and even parasites. However, high manufacturing costs have greatly limited their development as drugs, thus highlighting the need to develop novel and competitive production strategies. Here, a cost-effective procedure was established to produce the high amounts of peptides required for basic and clinical research. Firstly, a novel culture medium was designed, which was found to support significantly higher cell densities and recombinant expression levels of peptides under test compared to conventional media. The procedure has been also efficiently scaled up by using a 5 L fermenter, while the costs have been lowered significantly by developing a successful auto-induction strategy, which has been found to support higher yields of target constructs and cell biomass compared to conventional strategies based on expression induction by IPTG. Interestingly, it was estimated that by increasing production scale from 100 to 1000 mg/batch, unit costs decreased strongly from 253 to 42 (sic)/mg. These costs appear highly competitive when compared to chemical synthesis strategies. Altogether, the data indicate that the strategy represents an important starting point for the future development of large-scale manufacture of HDPs.
引用
收藏
页码:39 / 48
页数:10
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