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:10/19/2014)... October 19, 2014 According to ... (Access Points; WLAN Controllers; Wireless gateways)]- Global Advancements, ... segments the global Outdoor Wi-Fi market with analysis ... report MarketsandMarkets also identifies the drivers and the ... the trends, opportunities and challenges. , Browse 65 ...
(Date:10/18/2014)... 2014 The Asia-Pacific Bio-based Advanced ... the bio-based advanced phase change material market in ... Browse through the TOC of the Asia-Pacific Bio-based ... an idea of the in-depth analysis provided. This ... the Asia-Pacific bio-based advanced phase change material market, ...
(Date:10/18/2014)... (PRWEB) October 18, 2014 The Asian ... in Asia with analysis and forecast of revenue. The ... to $712.3 million by 2018, at a CAGR of ... TOC of the Asian arthroscopic devices market, to get ... provides a glimpse of the segmentation of arthroscopic devices ...
(Date:10/18/2014)... The Asia-Pacific Liquid Crystal on Silicon (LCoS) display market ... with analysis and forecast of revenue. This market was ... to reach $380.0 million by 2018, at a CAGR ... the TOC of the Asia-Pacific LCoS display market report, ... This also provides a glimpse of the segmentation of ...
Breaking Biology Technology:Outdoor Wi-Fi Market (by Access Points, WLAN Controllers, Wireless gateways) Expected to Cross $37.2 Billion by 2018 - New Report by MarketsandMarkets 2Outdoor Wi-Fi Market (by Access Points, WLAN Controllers, Wireless gateways) Expected to Cross $37.2 Billion by 2018 - New Report by MarketsandMarkets 3Outdoor Wi-Fi Market (by Access Points, WLAN Controllers, Wireless gateways) Expected to Cross $37.2 Billion by 2018 - New Report by MarketsandMarkets 4The Asia-Pacific Bio-based Advanced Phase Change Materials Market is estimated to grow to $39.8 million by 2018 - New Report by MicroMarket Monitor 2The Asia-Pacific Bio-based Advanced Phase Change Materials Market is estimated to grow to $39.8 million by 2018 - New Report by MicroMarket Monitor 3The Asia-Pacific Bio-based Advanced Phase Change Materials Market is estimated to grow to $39.8 million by 2018 - New Report by MicroMarket Monitor 4The Asian arthroscopic devices market is estimated to grow to $712.3 million by 2018 - New Report by MicroMarket Monitor 2The Asian arthroscopic devices market is estimated to grow to $712.3 million by 2018 - New Report by MicroMarket Monitor 3The Asian arthroscopic devices market is estimated to grow to $712.3 million by 2018 - New Report by MicroMarket Monitor 4The Asia-Pacific Liquid Crystal on Silicon (LCoS) display market is expected to reach $380.0 million by 2018 - New Report by MicroMarket Monitor 2The Asia-Pacific Liquid Crystal on Silicon (LCoS) display market is expected to reach $380.0 million by 2018 - New Report by MicroMarket Monitor 3
... where bacteria and leukocytes (white blood cells) move toward ... a novel method for treating sepsis, a systemic and ... blood. A team of researchers at the Massachusetts Institute ... designed a branchlike system of microfluidic channels, 20 micrometers ...
... Institute researchers Jorge Otero-Millan, Stephen Macknik, and Susana Martinez-Conde ... Neuroscience in a compelling study into why illusions ... Joseph,s Hospital and Medical Center in Phoenix. , The ... of illusory motion in Akiyoshi Kitaoka,s striking visual illusion, ...
... Bristol have created artificial muscles that can be transformed ... remarkable camouflaging abilities of organisms such as squid and ... they believe could be used in ,smart clothing, to ... The study is published today, 2 May, ...
Cached Biology Technology:Barrow researchers unravel illusion 2Squid and zebrafish cells inspire camouflaging smart materials 2
(Date:10/15/2014)... WASHINGTON, N.Y. , Oct. 15, 2014 /PRNewswire/ ... technology solutions for home and community-based care, today ... benefits of implementing Sandata,s Santrax® Electronic Visit Verification™ ... Care Services is a home health company founded ... Texas . The ...
(Date:10/15/2014)... to an unexpected extent. The outbreak does not follow ... shows a new disease dynamic in regions, where it ... German National Academy of Sciences Leopoldina, acatech – the ... of the German Academies of Sciences and Humanities have ... In the statement the academies call for the following ...
(Date:10/15/2014)... 2014   Neurotechnology , a provider of ... of the VeriLook Surveillance 3.0 software development ... biometric face identification using live video streams from ... new version not only identifies faces in a ... while they are moving through the video frame. ...
Breaking Biology News(10 mins):Sandata Announces Case Study with Quality Care Services, Inc. 2Academies call for consequences from the Ebola virus epidemic 2Academies call for consequences from the Ebola virus epidemic 3Academies call for consequences from the Ebola virus epidemic 4Academies call for consequences from the Ebola virus epidemic 5VeriLook Surveillance 3.0 SDK Identifies Faces and Moving Objects, Differentiates Pedestrians from Other Moving Objects in Video Surveillance Systems 2VeriLook Surveillance 3.0 SDK Identifies Faces and Moving Objects, Differentiates Pedestrians from Other Moving Objects in Video Surveillance Systems 3
... A new study involving the University of Colorado at ... Colorado and other western states indicates the size and ... and flammability of houses built in the so-called wildland-urban ... the Sept. 4 print edition of the Proceedings of ...
... elk, he might give them a word of advice: avoid ... A biological scientist at the University of Alberta, Boyce analyzed ... predation between wolves and elk are more strongly influenced by ... that even though wolf and elk populations overlapped in many ...
... ANN ARBOR, Mich. A new generation of implanted heart-assisting ... survive, and thrive, until they receive a heart transplant, a ... 75 percent of patients stay alive for at least six ... also helped patients original hearts regain function and allowed their ...
Cached Biology News:Wolves find happy hunting grounds in Yellowstone National Park 2Device helps patients survive, regain function til transplant 2Device helps patients survive, regain function til transplant 3Device helps patients survive, regain function til transplant 4