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.
Article
in Chinese
| 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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pichia
/
Recombinant Proteins
/
Cloning, Molecular
/
Fermentation
/
Genetics
/
Lactoferrin
/
Metabolism
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2004
Type:
Article
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