Abstract
Different carrier molecules have been fused to antimicrobial polypeptides (AMPs) to facilitate recombinant protein expression and purification. Some of them have improved the stability of AMPs and reduced the toxicity to host cells, but most current strategies still have some problems to be solved such as poor yield, low purity, high expense, time-consumption, and difficulty in scaling-up. Here, we introduced the elastin-like polypeptides (ELPs) as a fusion partner to express an antimicrobial polypeptide halocidin18 (Hal18). By the reversible soluble-insoluble phase transition, 69 mg of the fusion protein were purified from 1 l of culture medium with the purity of nearly 95%. After cleavage with hydroxylamine, the ELP's tag was easily separated from Hal18 in the next round of inverse transition cycle and Hal18 (1.7 mg, approximately 1.9 kDa) was mainly found in the supernatant with a recovery of about 47% and purity of 60%. Antimicrobial activity showed that Hal18 had strong antimicrobial activity against Escherichia coli and Micrococcus luteus but weak activity against Pichia pastoris.
References
Jan 1, 1977·Methods in Enzymology·P Bornstein, G Balian
Jan 1, 1992·Progress in Biophysics and Molecular Biology·D W Urry
Nov 15, 1991·The Biochemical Journal·D AndersonsH Steiner
Feb 1, 1988·Journal of Protein Chemistry·D W Urry
Dec 28, 1984·Biochemical and Biophysical Research Communications·D W UrryK U Prasad
Mar 27, 1996·Biochemical and Biophysical Research Communications·S Hara, M Yamakawa
Sep 1, 1995·Bio/technology·L A Collins-RacieE R LaVallie
Nov 5, 1999·Nature Biotechnology·D E Meyer, A Chilkoti
Aug 4, 2001·Biotechnology Progress·D E MeyerA Chilkoti
Jan 25, 2002·Nature·Michael Zasloff
Jun 18, 2002·FEBS Letters·Woong Sik JangIn Hee Lee
Feb 13, 2003·Biopolymers·Bin Li, Valerie Daggett
Apr 18, 2003·Protein Expression and Purification·Vitaly S SkosyrevLeonid M Vinokurov
Jul 25, 2003·Antimicrobial Agents and Chemotherapy·Woong Sik JangIn Hee Lee
Dec 19, 2003·Protein Expression and Purification·Philippa J BarrellAnthony J Conner
Dec 19, 2003·Nucleic Acids Research·Zhe Wang, Guangshun Wang
Jun 5, 2004·Protein Expression and Purification·Xian C RaoFu Q Hu
Aug 6, 2004·Protein Expression and Purification·Ingrid CipákováVladimír Velebný
Nov 24, 2004·Protein Science : a Publication of the Protein Society·Kimberly Trabbic-CarlsonAshutosh Chilkoti
Sep 10, 2005·Applied and Environmental Microbiology·Quande WeiHyung Joon Cha
Oct 12, 2005·Protein Expression and Purification·Xing-jun FengFan Zhang
Dec 6, 2005·Protein Expression and Purification·Yifeng LiGuangshun Wang
Dec 13, 2005·Current Opinion in Immunology·Kelly L Brown, Robert E W Hancock
Mar 18, 2006·Biochimica Et Biophysica Acta·Ja-Young MoonA Ramamoorthy
Jun 3, 2006·Biotechnology Progress·Dominic C ChowAshutosh Chilkoti
Apr 5, 2007·Nature Protocols·Hermann Schägger
Jun 19, 2007·Protein Expression and Purification·Xiaoxia XuWenqing Zhang
Jun 7, 2008·Current Microbiology·G-Q WuZ-L Shen
Citations
Mar 23, 2011·BMC Biotechnology·Fan HuLi Xin Ma
Jul 1, 2014·Journal of Controlled Release : Official Journal of the Controlled Release Society·Sarah R MacEwan, Ashutosh Chilkoti
Sep 8, 2010·Biotechnology Advances·Sagheer A Onaizi, Susanna S J Leong
Apr 21, 2016·Journal of Controlled Release : Official Journal of the Controlled Release Society·Agnes YeboahFrancois Berthiaume
Apr 26, 2016·Biotechnology and Bioengineering·Agnes YeboahFrancois Berthiaume
Jul 27, 2011·Peptides·Eduardo TapiaGloria Arenas
Aug 4, 2016·Journal of Biotechnology·Daniel A SousaOctávio L Franco
Sep 4, 2015·Biotechnology and Bioengineering·Sezin YigitDavid L Kaplan
Mar 18, 2020·3 Biotech·Chun-Guang YangXue-Jun Hu
Aug 30, 2020·Antibiotics·Yunqi MaJang-Su Park
Jan 26, 2021·ACS Biomaterials Science & Engineering·André da CostaRaul Machado
Aug 8, 2021·Biopolymers·Markian S BahniukLarry D Unsworth
May 23, 2015·Bioconjugate Chemistry·José Carlos Rodríguez-CabelloFrancisco Javier Arias
Aug 10, 2017·Langmuir : the ACS Journal of Surfaces and Colloids·Binwu ZhaoCarol K Hall
Aug 14, 2020·Biomacromolecules·Sergio AcostaJosé Carlos Rodríguez-Cabello