Document Detail


Small-scale slow glucose feed cultivation of Pichia pastoris without repression of AOX1 promoter: towards high throughput cultivations.
MedLine Citation:
PMID:  24326738     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Recombinant protein synthesis in Pichia pastoris is generally controlled by the strong methanol inducible AOX1 promoter which is repressed by glucose and glycerol. In shake flasks, commonly one or two methanol pulses are added per day for induction. Such pulse feeding procedure leads to carbon starvation phases, which may enhance proteolytic activities and, therefore, cause product losses. Starvation between the methanol pulses could be avoided with a continuous enzymatic feed of glucose from a glucose-based polymer. The amount of glucose was low enough to prevent AOX1 repression by glucose. Energy and carbon were continuously supplied for cell maintenance resulting in significantly increased cell densities and product activities, as shown here at the example of a fungal lipase expressed in P. pastoris. A threefold improvement in measured product activity was obtained by applying enzymatic glucose feed and a further improvement was achieved by applying a defined mixture of ammonium compounds. The strategy described here simplifies the general procedure in shaken cultures by allowing the direct continuation of the cultivation from glucose to the methanol-based production phase without a medium change. It is easily applicable to multiwell plates and thus beneficial for high throughput applications.
Authors:
Johanna Panula-Perälä; Antti Vasala; Janne Karhunen; Heikki Ojamo; Peter Neubauer; Anu Mursula
Related Documents :
2196238 - Immunoglobulin g, complement factor c3 and lymphocytes in proliferative intraocular dis...
24890138 - Impact of rosiglitazone on body composition, hepatic fat, fatty acids, adipokines and g...
18524198 - Acute bilateral dense cortical cataracts as a first manifestation of juvenile diabetes ...
19936028 - Increased neuronal nitric oxide synthase activity in retinal neurons in early diabetic ...
11869488 - Glucose and insulin metabolism in twins: influence of zygosity and birth weight.
17287148 - Prevalence of nonalcoholic fatty liver disease in women with polycystic ovary syndrome.
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-12-12
Journal Detail:
Title:  Bioprocess and biosystems engineering     Volume:  -     ISSN:  1615-7605     ISO Abbreviation:  Bioprocess Biosyst Eng     Publication Date:  2013 Dec 
Date Detail:
Created Date:  2013-12-11     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101088505     Medline TA:  Bioprocess Biosyst Eng     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Bioprocess Engineering Laboratory, Department of Process and Environmental Engineering, University of Oulu, P.O. Box 4300, 90014, Oulu, Finland, johanna.panula-perala@oulu.fi.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  A rapid in situ immobilization of D-amino acid oxidase based on immobilized metal affinity chromatog...
Next Document:  Graphene oxide can induce in vitro and in vivo mutagenesis.