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1.
Chinese Journal of Biotechnology ; (12): 1682-1689, 2015.
Artigo em Chinês | WPRIM | ID: wpr-337467

RESUMO

The 1,095 bp gene encoding peroxidase from Coprinus cinereus was synthesized and integrated into the genome of Pichia pastoris with a highly inducible alcohol oxidase. The recombinant CiP (rCiP) fused with the a-mating factor per-pro leader sequence derived from Saccharomyces cerevisiae was secreted into the culture medium and identified as the target protein by mass spectrometry, confirming that a C. cinereus peroxidase (CiP) was successfully expressed in P. pastoris. The endoplasmic reticulum oxidoreductase 1 (Ero1) and protein disulfide isomerase (PDI) were co-expressed with rCiP separately and simultaneously. Compared with the wild type, overexpression of PDI and Erol-PDI increaseed Cip activity in 2.43 and 2.6 fold and their activity reached 316 U/mL and 340 U/mL respectively. The strains co-expressed with Erol-PDI was used to high density fermentation, and their activity reached 3,379 U/mL, which was higher than previously reported of 1,200 U/mL.


Assuntos
Coprinus , Meios de Cultura , Citoplasma , Fermentação , Glicoproteínas , Metabolismo , Espectrometria de Massas , Fator de Acasalamento , Oxirredutases atuantes sobre Doadores de Grupo Enxofre , Metabolismo , Peptídeos , Peroxidases , Pichia , Metabolismo , Isomerases de Dissulfetos de Proteínas , Metabolismo , Dobramento de Proteína , Saccharomyces cerevisiae , Proteínas de Saccharomyces cerevisiae , Metabolismo
2.
Acta Physiologica Sinica ; (6): 417-432, 2013.
Artigo em Inglês | WPRIM | ID: wpr-297553

RESUMO

Fungal pathogens represent major problems for human health and agriculture. As eukaryotic organisms, fungi share some important features with mammalian cells. Therefore, current anti-fungal antibiotics often can not distinguish between fungi and mammalian cells, resulting in serious side effects in mammalian cells. Accordingly, there is strong impetus to develop antifungal alternatives that are both safe and effective. The E1 family of colicin are channel-forming bacteriocins produced by Escherichia coli, which are bactericidal only to E. coli and related species. To target the channel-forming domain of colicin to fungal cell membrane, we engineered a sexual mating pheromone of Candida albicans, α-factor pheromone to colicin Ia. A peptide was constructed consisting of an α mating pheromone of C. albicans fused to the channel-forming domain of colicin Ia to create a new fusion protein, pheromonicin-CA (PMC-CA). Indirect immunolabeling showed that the PMC-CA bound to fungal cells and inhibited growth in the laboratory and field. In the field, the protective activity of pheromonicin against rice blast disease was significantly greater, on a molar basis, than that of triazoles, tricyclazole or isoprothiolane. These results suggest that fusion peptides may be of value as fungicidal agents under agricultural conditions.


Assuntos
Candida albicans , Química , Colicinas , Química , Fungicidas Industriais , Química , Fator de Acasalamento , Peptídeos , Química , Engenharia de Proteínas
3.
J. venom. anim. toxins incl. trop. dis ; 15(1): 157-162, 2009. tab
Artigo em Inglês | LILACS, VETINDEX | ID: lil-508237

RESUMO

The frequency of spermatophore production and the amount of time between matings for laboratory-reared males of Tityus (Atreus) magnimanus to regenerate spermatophores were examined. Males attain sexual maturity at the fifth or sixth instar (after a period of 137 to 155 days) and can produce initial spermatophores shortly after maturation. After mating, males can regenerate spermatophores within a single 24-hour period and remate. The present contribution represents the first report on an aspect of reproductive biology in this species and is part of a continuing study of the life history and post-embryonic development of Tityus (Atreus) magnimanus.(AU)


Assuntos
Animais , Masculino , Espermatogônias , Desenvolvimento Embrionário , Escorpiões , Relatório de Pesquisa , Fator de Acasalamento
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