Producing human lactoferrin by high-density fermentation recombinant Pichia pastoris / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 181-185, 2004.
Artículo
en Chino
| WPRIM
| ID: wpr-281822
ABSTRACT
<p><b>BACKGROUND</b>To evaluate expression of human lactoferrin gene by high-density fermentation in recombinant Pichia pastoris on the premise of maintaining its biological activities.</p><p><b>METHODS</b>The neutrophil was isolated from human peripheral blood and its total RNA was prepared. Full-length cDNA of human lactoferrin gene was then obtained by RT-PCR, cloned into expression vector pPIC 3.5 K and transformed into Pichia pastoris strain KM71. With two-layer filter method, the transformants with high-productivity of human lactoferrin were screened out into fed-batch high-density fermentation. And later, the physical, chemical and biological activities of fermentation product were detected preliminarily.</p><p><b>RESULTS</b>The strain p3.5-k-7 with better productivity of human lactoferrin was screened out into fed-batch high-density fermentation. The fermentation lasted nearly for nine days, with A-600 of culture once above 260 and the highest productivity of human lactoferrin being 115 mg/L, 7.67 times the amount of that in shake flask cultivation.</p><p><b>CONCLUSION</b>The authors successfully realized high-density fermentation expression of human lactoferrin gene in recombinant Pichia pastoris.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Pichia
/
Proteínas Recombinantes
/
Clonación Molecular
/
Fermentación
/
Genética
/
Lactoferrina
/
Metabolismo
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Experimental and Clinical Virology
Año:
2004
Tipo del documento:
Artículo
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