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:4/19/2017)... Indiana (PRWEB) , ... April 18, 2017 , ... Alisa ... 2017 Distinguished Alumni Awards from the Purdue College of Pharmacy in Lafayette, Indiana. ... Purdue Pharmacy Program for achievements in their careers and other scientific endeavors. , ...
(Date:4/19/2017)... , ... April 19, 2017 , ... ... Series A-1 financing round. This event adds to several other early achievements at ... Executive and Scientific Teams. , ThermaGenix will use proceeds from the ...
(Date:4/19/2017)... and PUNE, India , April 19, 2017 ... Microfiltration Market: Global Opportunity Analysis and Industry Forecast, 2014-2022 ," the global market ... million by 2022, registering a CAGR of 9.6% from 2016 to 2022. ... ... ...
(Date:4/19/2017)... FRANCISCO, Calif. , April 19, 2017 /PRNewswire/ ... VCYT ) today announced that it will report ... of market on Wednesday, May 3, 2017. Following the ... webcast at 4:30 p.m. Eastern Time to discuss the company,s ... live webcast and subsequent replay may be accessed ...
Breaking Biology Technology:
(Date:3/22/2017)... -- Vigilant Solutions , a vehicle location and ... today the appointment of retired FBI special agent ... development. Mr. Sheridan brings more than 21 ... on the aviation transportation sector, to his new role ... served as the Aviation Liaison Agent Coordinator (ALAC) in ...
(Date:3/16/2017)... 2017 CeBIT 2017 - Against identity fraud with DERMALOG solutions "Made ... ... Used combined in one project, multi-biometric solutions provide a crucial contribution against identity ... Used combined in one project, multi-biometric ... ...
(Date:3/9/2017)... and MOUNTAIN VIEW, Calif. ... "Eating Well Made Simple," and 23andMe , the ... guide better food choices.  Zipongo can now provide customers ... food preferences, health goals and biometrics, but also genetic ... food choices. Zipongo,s personalized food decision support ...
Breaking Biology News(10 mins):