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:8/28/2014)... Ca. (PRWEB) August 28, 2014 Best ... rating for an alcohol-based hand sanitizer, asks food processors ... by comparing the hand sanitizer they’re currently using to ... . Hand hygiene is critical to fighting cross-contamination and ... Best Sanitizers believes there are key criteria that ...
(Date:8/28/2014)... PA (PRWEB) August 28, 2014 The ... create a culture that will allow individuals to perform ... becoming increasingly important for employers looking to compete in ... industries are embracing this trend, as the sector increasingly ... of a company, which will have real results on ...
(Date:8/28/2014)... Exciting new work by a Florida State University ... that can take your temperature, emit white light, and ... the molecule looks like a butterfly. , Biwu Ma, ... Engineering in the FAMU-FSU College of Engineering, created the ... has continued to discover that his creation has many ...
(Date:8/28/2014)... Diego, CA (PRWEB) August 28, 2014 ... Pipette.com is offering a promotion for Eppendorf ... Pipette.com customers to purchase the highest quality tubes ... Tubes are ideal for customers doing chemical, medical, ... lab technicians who are not regularly able to ...
Breaking Biology Technology:Food Processors and Food Handlers are Encouraged to Prepare for Fall Harvest by Evaluating Their Current Hand Sanitizer Using Nine Key Criteria 2Food Processors and Food Handlers are Encouraged to Prepare for Fall Harvest by Evaluating Their Current Hand Sanitizer Using Nine Key Criteria 3Klein Hersh International Leads Philadelphia Area Life Science Industry in Company Culture 2Klein Hersh International Leads Philadelphia Area Life Science Industry in Company Culture 3Novel 'butterfly' molecule could build new sensors, photoenergy conversion devices 2Eppendorf Safe Lock Centrifuge Tube Promotion Now Available at Major Pipette Distributor Pipette.com 2Eppendorf Safe Lock Centrifuge Tube Promotion Now Available at Major Pipette Distributor Pipette.com 3
... ... Area and Beyond, ATLANTA and BALTIMORE, ... Medicine is adding an Elekta,Synergy(R) S system to its Leksell Gamma ... cancer with,both intracranial and extracranial resources for whole-body cancer,treatment with the ...
... MONICA, Calif., Aug. 17 The Foundation,for Taxpayer and ... next meeting of the fledgling Interstate Alliance for Stem ... process builds,support for publicly funded science. The decision came ... a protest from FTCR. Warren Wollschlager, of the ...
... against osteoporosis-related fractures, including spine and hip, - Reclast reduced spine ... placebo in study published in New England Journal of Medicine, - Unique once-yearly ... ... - Osteoporotic fractures affect one in two women over 50(1) and are ...
Cached Biology Technology:Johns Hopkins to add Elekta Synergy S(R) to Cancer Treatment Lineup 2Johns Hopkins to add Elekta Synergy S(R) to Cancer Treatment Lineup 3Consumer Advocates Praise Interstate Stem Cell Open Meeting Decision 2Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 2Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 3Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 4Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 5Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 6
(Date:8/28/2014)... German . The ... a striking pattern of blue stripes alternating with golden stripes. ... and yellow cells emerge during growth in the skin of ... to compose the characteristic colour pattern. ,While it ... to form proper stripes, the embryonic origin of the pigment ...
(Date:8/28/2014)... Access to Research Careers) Program has announced the ... America,s 27th Annual Mouse Molecular Genetics Conference from ... Grove, California. These awards are meant to ... scientists from underrepresented groups into the mainstream of ... participation of young scientists at the Genetics Society ...
(Date:8/28/2014)... MARC (Maximizing Access to Research Careers) Program has ... College of Sports Medicine,s Conference on Integrative Physiology ... Florida. These awards are meant to promote ... from underrepresented groups into the mainstream of the ... of young scientists at the American College of ...
Breaking Biology News(10 mins):How the zebrafish gets its stripes 2How the zebrafish gets its stripes 3
... This news release is available in ... a major food staple for about 400 million people in the ... banana yields worldwide are severely threatened by pests. Dirk Hlscher from ... an international team of researchers have discovered that some banana varieties ...
... Shenzhen, China - Chinese researchers from Rui-Jin Hospital, ... other institutes identified the recurrent T372R mutation in ... related with insulinoma oncogenesis, implicating a potential marker ... neuroendocrine tumors (PNETs). The latest study was published ...
... protein deacylases has received considerable attention in recent years ... metabolic regulation. In a new study published in the ... Buck Institute researchers have now identified widespread regulation of ... Using a novel quantitative proteomic method developed at ...
Cached Biology News:Toxic substances in banana plants kill root pests 2Toxic substances in banana plants kill root pests 3Scientists identified T372R mutation as potential target for diagnosis and treatment of insulinoma 2SIRT5 regulation has dramatic effect on mitochondrial metabolism 2SIRT5 regulation has dramatic effect on mitochondrial metabolism 3
... Storage: The kit is stable for 12 months ... contents of the kit cold. , ,Principle: ... silica-gel based membrane which selectively adsorbs up to ... specialized binding buffers. Nucleotides, oligos ( , ,Application: ...
The Econo gradient pump rack is a preassembled rack that is used to hold the components of the Econo gradient pump kits....
... Leu DAP FLISP Assay with FAM-Labeled ... nm) • Accurately measure serine protease ... permeabilization • B-Bridge's FLISP Kits utilize ... proteases (FLISPs) to measure chymotrypsin-like activity ...
... instrument is the cycler that revolutionized ... uniformity and swappable blocks ensure consistent ... This rugged machine is more than ... for new features and upgrades as ...
Biology Products: