Navigation Links
Rapamycin Optimized in 100 Liter Fermentor Using BioCommand Control Strategys


Researchers in New Brunswick Scientific's in-house labs have developed a unique control strategy to achieve high antibiotic expression levels in a pilot-scale fermentation process of Streptomyces hygroscopicus. Using the BioFlo 6000 fermentor's on-line controller with touchscreen interface augmented by the powerful supervisory capabilities of AFSBioCommand software, scientists were successful in optimizing the culture environment for rapamycin production. Harvested dry cell weight recovered from the 100 liter fermentor was over 25 grams/L; intracellular rapamycin production was over 110 milligrams/L; and fermentation time was reduced by almost a day to 120 hours.


Introduction
Streptomyces expresses a variety of secondary metabolites, such as rapamycin, which are used in medical and biological applications. During fermentation, dense mycelia are produced, which are highly variable in phenotype and especially susceptible to changing conditions during cultivation. Formation of secondary metabolites is usually favored by sub-optimal cell growth conditions. Nutrient transfer problems can restrict the metabolism of the culture, and a high oxygen transfer rate (OTR) is always required. Therefore, it was necessary to develop a fermentation protocol which allowed for initial cell growth to a sufficient density as to facilitate the production of large quantities of the antibiotic, while preventing the culture from becoming so dense as to interfere with its ability to obtain vital nutrients and oxygen for the production of rapamycin.


Materials and Methods
Seed inoculum of Streptomyces hygroscopicus NBS-9746, selected from ATCC 29253 cell strain, was cultured in an Innova 4300 incubator shaker, then transferred to a 10 L BioFlo IV bench fermentor, and fina lly to a BioFlo 6000 Mobile Pilot Plant fermentor equipped with an ML-6100 touch-screen controller and 100 L working volurne vessel, (all equipment manufactured by New Brunswick Scientific, Edison, NJ). NBS' proprietary AFS-BioCommand software was used to control a multi-loop cascade strategy, wherein each of four process parameters were altered in sequence so as to maintain the desired D.O. level. Capable of managing up to eight fermentors, bioreactors, or ancillary instruments, this software automates the tedious and error-prone jobs of logging data, computing real and derived process values, and altering setpoints on the basis of time or metabolic events. In this study, AFS-BioCommand was programmed to maintain a D.O. level of 30% of air saturation by automatically increasing agitation rates from 200 to 450 rpm, then vessel pressure from 0.35 to 0.75 bar, aeration rate from 0.5 to 1 VVM and simultaneously shifting temperature from 28C to 25C, when D.O. fell below its setpoint. Based on optimization studies conducted on the bench level, pH was maintained between 6.2 and 6.8. A deadband was easily configured by using the local controller for the low setpoint and the supervisory control software for the high setpoint.


Discussion
Utilizing the multi-loop cascade strategy provided several benefits. First and foremost, D.O. was able to be maintained at optimal rates for production of rapamycin, a feat unable to be accomplished using a single loop control strategy ( see figure ). Agitation rates could also be significantly reduced, to minimize shear forces and enable higher yields of both cells and secondary metabolites.

Temperature was also lowered from that optimal for culture growth to the optimal range for rapamycin production. The lower temperature had the side benefit of slowing cell growth, thereby decreasing oxygen demands, which h elped to maintain the high D.O. levels.

For details of this study and further information on our bioprocessing supervisory software, speak with your local NBS representative. To discuss how you can put the expertise of our in-house researchers to work on your next process development, optimization, or contract production project, contact Dr. Julia Cino at 800-631-5417, by fax to 732-287-4222, or via e-mail at cino@nbsc.com.



'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Optimized Imaging of Protein Gels Stained with Coomassie Brilliant Blue Dye
2. Eagle Eye Optical Filter Optimized for Imaging Silver-Stained Gels
3. Optimized Dengue RT-PCR
4. P. pastoris Fermentation using a BioFlo 110 Benchtop Fermentor
5. E. coli Fermentation Using a BioFlo 110 Bench-Top Fermentor
6. An Easy-to-Use Research Fermentor/Bioreactor
7. Fundamentals of Fermentation: Techniques For Benchtop Fermentors
8. Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
9. Oil-Free PCR Using the Hot Top Assembly
10. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
11. Signal Transduction Reporting Systems Using Cis-Acting Enhancer Elements
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/23/2016)... Md. , June 23, 2016 A person ... from the crime scene to track the criminal down. ... the U.S. Food and Drug Administration (FDA) uses DNA evidence ... Sound far-fetched? It,s not. The FDA ... sequencing to support investigations of foodborne illnesses. Put as simply ...
(Date:6/23/2016)... Cancer Foundation (PCF) is pleased to announce 24 new Young Investigator (YI) ... of the Class of 2016 were selected from a pool of 128 applicants ... the Class of 2016 PCF Young Investigators ... ... ...
(Date:6/23/2016)... ... June 23, 2016 , ... Supplyframe, the Industry Network for ... Design Lab . Located in Pasadena, Calif., the Design Lab’s mission is to ... are designed, built and brought to market. , The Design Lab is Supplyframe’s ...
(Date:6/23/2016)... June 23, 2016 Apellis Pharmaceuticals, Inc. ... clinical trials of its complement C3 inhibitor, APL-2. ... multiple ascending dose studies designed to assess the ... subcutaneous injection in healthy adult volunteers. ... as a single dose (ranging from 45 to ...
Breaking Biology Technology:
(Date:5/24/2016)... , May 24, 2016 Ampronix facilitates superior patient care by providing ...  3D medical LCD display is the latest premium product recently added to the range ... ... ... Sony 3d Imaging- LCD Medical Display- Ampronix News ...
(Date:5/3/2016)... , May 3, 2016  Neurotechnology, a ... the MegaMatcher Automated Biometric Identification System (ABIS) ... large-scale multi-biometric projects. MegaMatcher ABIS can process multiple ... using any combination of fingerprint, face or iris ... MegaMatcher SDK and MegaMatcher Accelerator , ...
(Date:4/19/2016)... The new GEZE SecuLogic access ... "all-in-one" system solution for all door components. It can ... door interface with integration authorization management system, and thus ... minimal dimensions of the access control and the optimum ... offer considerable freedom of design with regard to the ...
Breaking Biology News(10 mins):