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:7/23/2014)... Quantum Center at Skolkovo) and Mario Ziman (Masaryk ... of Physics in Bratislava, Slovakia) have found a ... through an amplifier and, conversely, when transmitting a ... an article published in the journal Physical ... particles are considered to be the basis of ...
(Date:7/23/2014)... Last year, Formedix celebrated its partnership ... of data management solutions for clinical trials. The company ... electronic data capture (EDC) and clinical data management. Formedix, ... tools added TrialMaster to its Transform for EDC product ... for TrialMaster, is being released this week, in response ...
(Date:7/23/2014)... MOINES, Iowa (PRWEB) July 23, 2014 ... join the business as vice president of Agricultural Biotechnology ... Mendel Biotechnology where he served most recently as president, ... extensive biotechnology and business leadership in the seed and ... Pioneer,” said Paul E. Schickler , president of ...
(Date:7/23/2014)...  Sofinnova Ventures, a Menlo Park ... Sofinnova Venture Partners IX, L.P. at the hard cap ... fund target of $425 million. Sofinnova focuses ... with recent funds, SVP IX will provide capital primarily ... with select investments in earlier stage opportunities. Investments will ...
Breaking Biology Technology:Scientists find way to maintain quantum entanglement in amplified signals 2Scientists find way to maintain quantum entanglement in amplified signals 3Scientists find way to maintain quantum entanglement in amplified signals 4Formedix and OmniComm Systems® Partnership Strengthened with the Release of Enhanced TrialMaster® Integration for EDC Build 2Formedix and OmniComm Systems® Partnership Strengthened with the Release of Enhanced TrialMaster® Integration for EDC Build 3DuPont Pioneer Names Neal Gutterson to Lead Agricultural Biotechnology 2DuPont Pioneer Names Neal Gutterson to Lead Agricultural Biotechnology 3Sofinnova Raises $500 Million Biotech Venture Fund 2
... The single-atom thick material graphene maintains its high ... step to advancing the material from a laboratory phenomenon ... devices, researchers report in the April 9 issue of ... engineers and theoretical physicists from the University of Texas ...
... Mechanical Engineering at Stevens Institute of Technology was named ... (YIP) award by the Office of Naval Research (ONR). ... exceptional promise for creative study. "We are proud ... said ONR,s Director of Research Dr. Michael Kassner. "Through ...
... ... Regulatory Submissions , ... Rockville, MD (Vocus) April 8, 2010 -- Bringing its popular interactive workshop to Europe ... eCTD Submissions in Brussels, 5–7 May. This introductory workshop guides participants on how to ...
Cached Biology Technology:With support, graphene still a superior thermal conductor 2Dr. Chang-Hwan Choi honored with prestigious Young Investigator Award 2Dr. Chang-Hwan Choi honored with prestigious Young Investigator Award 3RAPS to Host eCTD Submissions Workshop in Brussels 2RAPS to Host eCTD Submissions Workshop in Brussels 3
(Date:7/23/2014)... the world have no access to clean drinking water. ... people die from water-related diseases every year. ETH student ... contribution to solving this problem. Working with researchers from ... Functional Materials Laboratory, the 23-year-old spent a year researching ... makes our DrinkPure filter unique is that you can ...
(Date:7/23/2014)... personalized cancer medicine, (the WIN consortium), recently announced that ... University will be one of its six new ... in the Tri-State Area. The goal of WIN is ... the patient by forging collaborations between partners in academia, ... WIN consortium means Jefferson cancer patients will have access ...
(Date:7/23/2014)... potential for clinical use of induced pluripotent stem ... previously been hindered by the risk of dysregulated ... ability to use etoposide treatment to halt teratoma ... disease, specifically acute myocardial infarction, is demonstrated in ... , a peer-reviewed journal from Mary Ann Liebert, ...
Breaking Biology News(10 mins):ETH student develops filter for clean water around the world 2New partnership brings more personalized cancer treatment to Philadelphia 2
... research professor and professor emeritus at The University of ... the American Academy of Arts & Sciences (AAAS), along ... Earl Jones, U.S. Secretary of Defense Robert Gates and ... scientist who translates his discoveries into technology that helps ...
... New York Sea Grant (NYSG) announces the release of ... Populations in South Shore Bays of Long Island, NY, ... of five research projects funded through NYSG,s Hard Clam ... NOAA National Marine Fisheries Service, South Shore Estuary Reserve, ...
... In Maine, government scientists have figured out how to measure ... and the sun - all the while saving money, energy, ... this energy-efficient ingenuity was the need to help the National ... spring snowmelt. The problem was this: While the USGS ...
Cached Biology News:University of Texas at Austin engineer elected to American Academy of Arts and Sciences 2University of Texas at Austin engineer elected to American Academy of Arts and Sciences 3Sea Grant report synthesizes recent research on New York's clams 2Measuring snow with a bucket, a windmill, and the sun? 2
... Expansion Kit - Neurosphere is a system designed ... expansion as neurospheres in a serum free environment. ... Media Supplement, which has been optimized for neural ... . (1996) Genes & Development 10 :3129). ...
Request Info...
... RQ1 RNase-Free DNase is a ... single-stranded or double-stranded DNA to produce ... for use in applications where maintaining ... RQ1 DNase is qualified for use ...
... Polymerase I catalyzes the template-directed polymerization of ... direction. DNA Polymerase I possesses a 3?5 ... the error rate during DNA replication, and ... enables the enzyme to replace nucleotides in ...
Biology Products: