An improved and simplified method for the large-scale purification of pediocinPA-I produced by Pediococcus acidilactici

被引:17
作者
Beaulieu, L
Aomari, H
Groleau, D
Subirade, M
机构
[1] Univ Laval, STELA, Ctr INAF, Canada Res Chair Prot Biosyst & Funct Foods Nutra, Ste Foy, PQ G1K 7P4, Canada
[2] Natl Res Council Canada, Biotechnol Res Inst, Montreal, PQ H4P 2R2, Canada
关键词
bacteriocin; hydrophobic interaction; pediocin PA-I; Pediococcus acidilactici PAC 1.0; purification; scalable method;
D O I
10.1042/BA20050041
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The bacteriocin pediocin PA-I produced by Pediococcus acidilactici PAC 1.0 offers significant potential as a food preservative and as an antimicrobial agent in the medical area. However, low production yields and difficulties in obtaining significant amounts of pure pediocin PA-1 have limited, in part, its biochemical and physical characterization. In the present study, we describe a simple and more efficient purification strategy for pediocin PA-1. A hydrophobic interaction chromatography step using an octyl-Sepharose column was introduced for final purification and polishing. The new method is a scalable one, uses only two steps and yields highly purified pediocin PA-I with a recovery as high as 73%, which is at least two to three times more than that of the methods reported so far. Highly purified, biologically active pediocin PA-1 of the correct molecular mass (4624 Da, with two disulphide bridges) was obtained. Fourier-transform infrared analysis runs at p(2)H 6 indicated that pediocin PA-1 was more structured than similar pediocin PA-1 samples purified using the earlier purification scheme.
引用
收藏
页码:77 / 84
页数:8
相关论文
共 27 条
[1]   QUANTITATIVE STUDIES OF THE STRUCTURE OF PROTEINS IN SOLUTION BY FOURIER-TRANSFORM INFRARED-SPECTROSCOPY [J].
ARRONDO, JLR ;
MUGA, A ;
CASTRESANA, J ;
GONI, FM .
PROGRESS IN BIOPHYSICS & MOLECULAR BIOLOGY, 1993, 59 (01) :23-56
[2]   DIRECT DETECTION OF AN ANTIMICROBIAL PEPTIDE OF PEDIOCOCCUS-ACIDILACTICI IN SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS [J].
BHUNIA, AK ;
JOHNSON, MC ;
RAY, B .
JOURNAL OF INDUSTRIAL MICROBIOLOGY, 1987, 2 (05) :319-322
[3]   The lantibiotic nisin, a special case or not? [J].
Breukink, E ;
de Kruijff, B .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1999, 1462 (1-2) :223-234
[4]   EXAMINATION OF THE SECONDARY STRUCTURE OF PROTEINS BY DECONVOLVED FTIR SPECTRA [J].
BYLER, DM ;
SUSI, H .
BIOPOLYMERS, 1986, 25 (03) :469-487
[5]   Expanded bed adsorption as a unique unit operation for the isolation of bacteriocins from fermentation media [J].
Callewaert, R ;
De Vuyst, L .
BIOSEPARATION, 1999, 8 (1-5) :159-168
[6]   Purification of bacteriocins of lactic acid bacteria: Problems and pointers [J].
CarolissenMackay, V ;
Arendse, G ;
Hastings, JW .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 1997, 34 (01) :1-16
[7]   Comparative antimicrobial activity of enterocin L50, pediocin PA-1, nisin A and lactocin S against spoilage and foodborne pathogenic bacteria [J].
Cintas, LM ;
Casaus, P ;
Fernandez, MF ;
Hernandez, PE .
FOOD MICROBIOLOGY, 1998, 15 (03) :289-298
[8]   FOURIER-TRANSFORM INFRARED SPECTROSCOPIC INVESTIGATION OF PROTEIN STABILITY IN THE LYOPHILIZED FORM [J].
COSTANTINO, HR ;
GRIEBENOW, K ;
MISHRA, P ;
LANGER, R ;
KLIBANOV, A .
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1995, 1253 (01) :69-74
[9]   Conformational changes of pediocin in an aqueous medium monitored by Fourier transform infrared spectroscopy: a biological implication [J].
Gaussier, H ;
Lavoie, M ;
Subirade, M .
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 2003, 32 (1-2) :1-9
[10]   Replacement of trifluoroacetic acid with HCl in the hydrophobic purification steps of pediocin PA-1: A structural effect [J].
Gaussier, H ;
Morency, H ;
Lavoie, MC ;
Subirade, M .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (10) :4803-4808