Navigation Links
High Yield Protein Production from Pichia pastoris Yeast: A Protocol for Benchtop Fermentation

ed before the culture was switched to the production phase.

The production phase (methanol feeding) was started after 43 hours of cell growth. The production feed medium consists of 100% methanol and 12 mL/L trace metal solution. Feeding rates were divided into three stages: 6 hr induction, 48 hr in a high-feed-rate stage and 44 hr in a low-feed-rate stage. The feeding rate of the induction stage was ramped from 1 to 10.9 mL/L/hr, which was controlled by the computer program. Feeding rates in the high and low rate stages were 15 and 2 mL/L/hr respectively. The total volume of feed was 2 L. During the fermentation, oxygen demand can be quite high and oxygen was added to the air stream automatically. (Figure 2)

pH is usually adjusted to inhibit the activities of proteinases existing in the culture broth during the production phase. Furthermore, since a host strain that is protease deficient was used, it was not necessary to change the pH level when culture was shifted from cell growth phase to production phase. It has been found that pH 5 is the optimal for cell metabolism and cell growth and that the oxygen consumption rate is higher at that pH. Using this protocol, optical densities of up to 630 can be obtained. Protein expression, of course, varies with the particular protein being expressed.

Although not without problems related to its culture, P. pastoris culture protocols are scalable and have become a powerful tool for the production of commercially valuable proteins.

More information on P. pastoris culture (expression vectors, protocols, etc.) can be obtained from Invitrogen, Inc., Carlsbad, CA and from Pichia Protocols published by Humana Press and edited by David R. Higgins and James M. Cregg



Figure 1: BioFlo 3000 fermentor with supervisory control system.


Figure 2: Pure oxygen s
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. Yeast Protein Production System Features High Yields and One-Step Purification
2. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
3. Combine High Yield, Great Accuracy and the Prevention of Carry-over Contamination by Using the Novel Expand High FidelityPLUS PCR System
4. ArrayScript RT: Maximize Yields of Full-length cDNA
5. Increase Protein Activity and Yield
6. High Yield Transcription for Every Application
7. Maximize Viral RNA Yield from Biological Fluid
8. Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue
9. Synthesize High Yields of Biotinylated aRNA
10. Translate High Yields of Active Protein
11. Translate High Yields of Biologically Active Proteins
Post Your Comments:
(Date:7/10/2014)... , July 10, 2014 Research ... addition of the "International Photonic Integrated Circuit ... Forecasts to 2019" report to their offering. ... The concept of photonic integration traces its ... century. The promise of photonic integration went unexplored ...
(Date:7/10/2014)... fight against terrorism with the creation of a sensor ... use of light and special glass fibres. , Published ... , the researchers describe a novel optical fibre sensor ... 6.3 ppm (parts per million). It requires an analysis ... detection has involved looking for metals that encase them ...
(Date:7/10/2014)... Understanding the basis of psychiatric disorders has been extremely ... increase risk but are insufficient to cause disease. Now ... Cell Press journal Cell Stem Cell describe ... genetic risks interact with other risk factors or environmental ... Their research pinpoints a genetic variant that may predispose ...
(Date:7/10/2014)... sit upside down forever. Nanophysics makes sure of that. Mechanics ... just published in Physical Review E . Knowledge that ... that seem to be able to sit still forever, and ... years. We will soon be able to buy smart new ... But the fact is, sooner or later the grip is ...
Breaking Biology Technology:International Photonic Integrated Circuit (Monolithic Integration, Hybrid Integration, Module Integration) Market - Forecasts to 2019 2Detecting trace amounts of explosives with light 2New strategy could uncover genes at the root of psychiatric illnesses 2Even geckos can lose their grip 2
... ATLANTA, Jan. 21 CryoLife, Inc. (NYSE: ... an industry milestone when it received tissue from its ... 2009 came from a Kansas-based organ and tissue procurement ... with the support of the organ and tissue procurement ...
... Jan. 21 , News ... Organizations spanning the life sciences industry -- ... manufacturers -- are increasingly turning to Oracle(R) ... more efficiently and effectively. , , ...
... January 21 eTrinsic, a division of Simbionix ... products and,technology for training medical salespeople and professionals, ... as Sales Executive for the,Midwest and Eastern Region. ... located in,Grayslake, Illinois. , ...
Cached Biology Technology:CryoLife, Inc. Receives Tissue From 100,000th Donor, Marking Industry Milestone 2Life Sciences Organizations Choose Oracle to Accelerate and Simplify Clinical Trial Data Collection 2Life Sciences Organizations Choose Oracle to Accelerate and Simplify Clinical Trial Data Collection 3Life Sciences Organizations Choose Oracle to Accelerate and Simplify Clinical Trial Data Collection 4Glen C. Williams Joins eLearning Sales Team for Simbionix, Heading up Chicago Office 2
(Date:7/11/2014)... to the hippocampus, the marginal division of the ... What is the impact degree of substance P ... memory function? Yan Yu and his team, College ... using immunofluorescence staining, that substance P receptor, neurokinin ... striatal marginal division of normal rats. Unilateral or ...
(Date:7/11/2014)... Janeiro, Brazil- In the brains of all vertebrates, ... allows an electric or chemical signal to be ... synapses, which are the most abundant type of ... formation is crucial for learning, memory, perception and ... synapses critical for brain function. For instance, ...
(Date:7/10/2014)... , July 1, 2014 Smart technology ... for convenience and improved security: NXT-ID, Inc. (OTCBB: NXTD), eBay ... GOOG ), Apple Inc. (NASDAQ: AAPL ), ... (NYSE: MA) NXT-ID, Inc., (OTCQB: NXTD) a biometric authentication ... announce that the second series of 30 second spots on ...
Breaking Biology News(10 mins):Blame it on the astrocytes 2Advantages for Digital Wallets Eliminate Need to Carry Credit Cards - Large Scale CNBC Ad Campaign for Latest Smart Wallet to Commence July 7th 2Advantages for Digital Wallets Eliminate Need to Carry Credit Cards - Large Scale CNBC Ad Campaign for Latest Smart Wallet to Commence July 7th 3Advantages for Digital Wallets Eliminate Need to Carry Credit Cards - Large Scale CNBC Ad Campaign for Latest Smart Wallet to Commence July 7th 4Advantages for Digital Wallets Eliminate Need to Carry Credit Cards - Large Scale CNBC Ad Campaign for Latest Smart Wallet to Commence July 7th 5
... most species seem well adapted to their environment. Scientists ... evolutionary changes occur at a surprisingly rapid pace. ... a new book by John N. Thompson, distinguished professor ... California, Santa Cruz. In Relentless Evolution (University of Chicago ...
... the usual way proteins are made may explain mysterious ... disabilities. The discovery, made by a team ... may lead to better treatments for older adults with ... Fragile X-associated Tremor Ataxia Syndrome (FXTAS), causes shakiness and ...
... has been presented today to Professor Adolfo Diez-Perez, MD, ... Hospital del Mar, Director of the Bone and Joint ... and Professor of Medicine at the Autonomous University of ... IOF Medal of Achievement every two years to honour ...
Cached Biology News:New book explores relentless evolution in a constantly changing world 2Science surprise: Toxic protein made in unusual way may explain brain disorder 2Science surprise: Toxic protein made in unusual way may explain brain disorder 3IOF Medal of Achievement awarded to Professor Adolfo Diez-Perez 2
SlowFade Antifade Kit with DAPI...
nuclear RNA export factor 1, mRNA...
The Captivate Disposable Sample Chamber is designed for use with the Captivate magnetic yoke assembly (C-24700) and can be used with our Captivate ferrofluid conjugates (C-21473 C-21474 C-21476) or ...
... analysis of gene expression) technology ... Expression profiles generated by SAGE ... low-abundance transcripts making it possible ... I-SAGE Kit is designed to ...
Biology Products: