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:12/19/2014)... 2014 iLab Solutions, the global leader ... Siri Bryant as the new Director of Institutional Implementations ... Implementations. These two new leadership positions were created to ... ensuring that iLab continues to meet the needs of ... years, iLab has been deployed at over 450 core ...
(Date:12/19/2014)... , Dec. 18, 2014  Decision Resources Group finds ... biomaterial market will grow moderately through 2023 as the ... growing awareness of dental biomaterials all spur procedure volumes. ... as China and ... dental procedures within reach for a larger proportion of ...
(Date:12/17/2014)... 16, 2014 The Activated Carbon ... which is why IBISWorld updated its original industry research ... benefit from an intensified focus on environmental policy. Over ... emission standards for power plants and a range of ... IBISWorld Industry Analyst Sarah Kahn, “a range of downstream ...
(Date:12/17/2014)... Ipsen Biopharmaceuticals, Inc., an affiliate ... IPSEY), today announced that Somatuline® Depot® (lanreotide) ... approved by the U.S. Food and Drug Administration ... (GEP-NETs) in adult patients with unresectable, well or ... improve progression-free survival (PFS). , “The approval of ...
Breaking Biology Technology:iLab Solutions Announces Two New Leadership Positions to Meet the Needs of Its Growing Customer Base 2Asia Pacific Dental Biomaterial Market Will Grow Strongly Through 2023 2Asia Pacific Dental Biomaterial Market Will Grow Strongly Through 2023 3Activated Carbon Manufacturing in the US Industry Market Research Report Has Been Updated from IBISWorld 2Activated Carbon Manufacturing in the US Industry Market Research Report Has Been Updated from IBISWorld 3Ipsen Announces the FDA Approval of Somatuline® Depot® (lanreotide) Injection for the Treatment of Gastroenteropancreatic Neuroendocrine Tumors 2Ipsen Announces the FDA Approval of Somatuline® Depot® (lanreotide) Injection for the Treatment of Gastroenteropancreatic Neuroendocrine Tumors 3Ipsen Announces the FDA Approval of Somatuline® Depot® (lanreotide) Injection for the Treatment of Gastroenteropancreatic Neuroendocrine Tumors 4Ipsen Announces the FDA Approval of Somatuline® Depot® (lanreotide) Injection for the Treatment of Gastroenteropancreatic Neuroendocrine Tumors 5Ipsen Announces the FDA Approval of Somatuline® Depot® (lanreotide) Injection for the Treatment of Gastroenteropancreatic Neuroendocrine Tumors 6Ipsen Announces the FDA Approval of Somatuline® Depot® (lanreotide) Injection for the Treatment of Gastroenteropancreatic Neuroendocrine Tumors 7Ipsen Announces the FDA Approval of Somatuline® Depot® (lanreotide) Injection for the Treatment of Gastroenteropancreatic Neuroendocrine Tumors 8
... Feb. 6, 2012  Auxilium Pharmaceuticals, Inc. (Nasdaq: ... announced the appointment of two executive officers, effective ... Vice President, Chief Administrative Officer and General Counsel, ... Vice President, Sales. "The leadership appointments ...
... 2012 Ikaria, Inc., a critical care company focused on ... needs of critically ill patients in the hospital and ICU ... will present at the 14th Annual BIO CEO & Investor ... 13, 2012 at 3:30 p.m. EST in the Park South ...
... , a Manchester team lead by Nobel laureates Professor Andre ... dimension in graphene research. Their research shows a transistor that ... next silicon. Graphene one atomic plane of ... from electronic to chemical and from optical to mechanical. ...
Cached Biology Technology:Auxilium Announces Appointments to Enhance Leadership Team 2Auxilium Announces Appointments to Enhance Leadership Team 3Auxilium Announces Appointments to Enhance Leadership Team 4Auxilium Announces Appointments to Enhance Leadership Team 5Ikaria® to Present at 14th Annual BIO CEO & Investor Conference 2Graphene electronics moves into a third dimension 2
(Date:12/10/2014)... N.C. , Dec. 9, 2014  Valencell, a ... it is seeing a staggering demand from its licensees ... the Company revealed, is not solely coming from fitness ... as well. "A wearable is only as ... result, accuracy is the ultimate driver in long-term mass ...
(Date:12/10/2014)... , Dec. 08, 2014 Research and Markets ... addition of the "Biometrics Market in Japan ... http://photos.prnewswire.com/prnh/20130307/600769 The ... such as rural banking and upgradation of the ... witnessed in the market. Besides the aforementioned projects, ...
(Date:12/5/2014)... 4, 2014  Tute Genomics, a leader in cloud-based ... Series A1 funding led by UK-based Eurovestech. Peak Ventures ... the investment round. "We are at a ... next-generation sequencing and seeks new approaches for the diagnosis, ... , MD MBA, and CEO of Tute Genomics. "One ...
Breaking Biology News(10 mins):Wearable Technology Products Demand Highly Accurate Biometric Technology 2Biometrics Market in Japan 2014-2018: Key Vendors are DDS, Fujitsu, Hitachi and NEC 2Tute Genomics Raises $2.3 Million to Scale Up Cloud-Based Genome Analytics and Accelerate Personalized Medicine 2Tute Genomics Raises $2.3 Million to Scale Up Cloud-Based Genome Analytics and Accelerate Personalized Medicine 3
... a gene implicated in human cancers, researchers from Whitehead Institute ... online issue of Science. , MicroRNAs are tiny snippets of ... the gene's messenger RNA (which copies DNA information and starts ... microRNA was discovered in 1993, in worms. It took seven ...
... digest cellulose, but the human process of producing ethanol ... is the sluggish rate at which the cellulose enzyme ... are then fermented into ethanol. , To help unlock ... molecular simulations at the San Diego Supercomputer Center (SDSC), ...
... parents, 8 million cases of acute middle ear infections every ... trips to the pediatrician. But new research from a collaboration ... all that. , In the March 23 issue of ... Jude show that middle ear infections in mice can be ...
Cached Biology News:MicroRNA helps prevent tumors 2MicroRNA helps prevent tumors 3Meeting the ethanol challenge: Scientists use supercomputer to target cellulose bottleneck 2Meeting the ethanol challenge: Scientists use supercomputer to target cellulose bottleneck 3Viral protein is an effective preventative against infection 2
Mouse monoclonal antibody raised against a partial recombinant QARS. NCBI Entrez Gene ID = QARS...
96-well format (For processing samples on microplate-size MALDI target plates such as those from Bruker, Shimadzu, Thermo, and PerkinElmer) Packaging: 2 / pack...
... mixing devices. Stepping motors device is maintenance ... durable shaker to provide a uniform motion ... nature. Regulated action scale for all modes. ... control time, continuous time, action scale and ...
Mouse monoclonal antibody raised against a partial recombinant STAU2. NCBI Entrez Gene ID = STAU2...
Biology Products: