Abstract:
:We have evaluated the cloning and functional expression of previously described broad antimicrobial spectrum bacteriocins SRCAM 602, OR-7, E-760, and L-1077, by recombinant Pichia pastoris. Synthetic genes, matching the codon usage of P. pastoris, were designed from the known mature amino acid sequence of these bacteriocins and cloned into the protein expression vector pPICZαA. The recombinant derived plasmids were linearized and transformed into competent P. pastoris X-33, and the presence of integrated plasmids into the transformed cells was confirmed by PCR and sequencing of the inserts. The antimicrobial activity, expected in supernatants of the recombinant P. pastoris producers, was purified using a multistep chromatographic procedure including ammonium sulfate precipitation, desalting by gel filtration, cation exchange-, hydrophobic interaction-, and reverse phase-chromatography (RP-FPLC). However, a measurable antimicrobial activity was only detected after the hydrophobic interaction and RP-FPLC steps of the purified supernatants. MALDI-TOF MS analysis of the antimicrobial fractions eluted from RP-FPLC revealed the existence of peptide fragments of lower and higher molecular mass than expected. MALDI-TOF/TOF MS analysis of selected peptides from eluted RP-FPLC samples with antimicrobial activity indicated the presence of peptide fragments not related to the amino acid sequence of the cloned bacteriocins.
journal_name
Biomed Res Intjournal_title
BioMed research internationalauthors
Arbulu S,Jiménez JJ,Gútiez L,Cintas LM,Herranz C,Hernández PEdoi
10.1155/2015/767183subject
Has Abstractpub_date
2015-01-01 00:00:00pages
767183eissn
2314-6133issn
2314-6141journal_volume
2015pub_type
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journal_title:BioMed research international
pub_type: 杂志文章
doi:10.1155/2014/309307
更新日期:2014-01-01 00:00:00
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pub_type: 杂志文章,评审
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pub_type: 临床试验,杂志文章
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pub_type: 杂志文章
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journal_title:BioMed research international
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journal_title:BioMed research international
pub_type: 杂志文章,多中心研究
doi:10.1155/2019/7041607
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journal_title:BioMed research international
pub_type: 杂志文章,随机对照试验
doi:10.1155/2014/632765
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doi:10.1155/2017/4276928
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journal_title:BioMed research international
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doi:10.1155/2014/802841
更新日期:2014-01-01 00:00:00
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journal_title:BioMed research international
pub_type: 杂志文章,评审
doi:10.1155/2015/682857
更新日期:2015-01-01 00:00:00
abstract:INTRODUCTION:The aim of this report is to study the graft and patient survival in a large cohort of recipients with an analysis of factors that may affect the final outcomes. METHODS:Between March 1976 and March 2008, 1967 consecutive live-donor renal transplants were carried out. Various variables that may have an im...
journal_title:BioMed research international
pub_type: 临床试验,杂志文章
doi:10.1155/2013/912413
更新日期:2013-01-01 00:00:00
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journal_title:BioMed research international
pub_type: 杂志文章
doi:10.1155/2020/1204605
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journal_title:BioMed research international
pub_type: 杂志文章
doi:10.1155/2018/5484731
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journal_title:BioMed research international
pub_type: 杂志文章
doi:10.1155/2013/276417
更新日期:2013-01-01 00:00:00
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pub_type: 杂志文章,评审
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doi:10.1155/2014/696475
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journal_title:BioMed research international
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