Enhancement and Scale-Up of ?-(1, 3) Glucan Production by Agrobacterium sp.

Author(s):  
Sri Lakshmi Puliga ◽  
Suhas Handa ◽  
Sathyanarayana N Gummadi ◽  
Mukesh Doble

Curdlan is a water insoluble polysaccharide composed exclusively of ?-(1, 3) linked glucose residues. Agrobacterium sp. is known to produce extracellular curdlan under nitrogen-limited conditions. The purpose of this study was to investigate the effects of pH, amounts of ammonium, sucrose and trace elements, and time the addition of sucrose, ammonium and uracil on the production of curdlan in a shake flask and to further scale-up the process to a 5 L fermentor. A maximum of 48.7 g L-1 of curdlan was obtained in a shake flask when 150 and 1.4 g L-1 of sucrose and ammonium were used at the initial pH of 6.5. The production was enhanced to 57 g L-1 by adding one third of sucrose and 1 g L-1 of uracil at the 48th h in a fed batch mode. The process was scaled up to a 5 L bioreactor in a batch mode where the oxygen transfer rate was higher (0.192 mg L-1s-1) when compared to that in the shake flask (0.096 mg L-1s-1). Curdlan production was 58 g L-1 in the bioreactor, which was higher than the shake flask under batch conditions (48.7 g L-1). The viscosity average molecular weight of the curdlan produced was found to be 1.4 × 105.

2003 ◽  
Vol 46 (3) ◽  
pp. 455-459 ◽  
Author(s):  
Georgina L. Michelena ◽  
Aidín Martínez ◽  
Antonio Bell ◽  
Emilia Carrera ◽  
Roxana Valencia

Fed batch fermentation was carried out for the dextransucrase enzyme production from Leuconostoc mesenteroides and the production was scale-up using oxygen transfer criteriuom. It was found that in 5 L vessel fermentation capacity, the best agitation speed was 225 min-1 and aeration rate was 0.15 vvm, obtaining dextransucrase activity of 127 DSU/mL.. The maximum enzyme production velocity coincide with the maximum growth velocity between 6 and 7 h of fermentation, which confirmed that dextransucrase production was associated with microbial growth. High enzyme yields were achieved during scale up based on oxygen transfer rate.


2012 ◽  
Vol 69 ◽  
pp. 182-195 ◽  
Author(s):  
Cornelia Bähr ◽  
Bernd Leuchtle ◽  
Christian Lehmann ◽  
Julia Becker ◽  
Markus Jeude ◽  
...  

Author(s):  
Nina Ihling ◽  
Lara Pauline Munkler ◽  
Christoph Berg ◽  
Britta Reichenbächer ◽  
Johannes Wirth ◽  
...  

Cultivations of mammalian cells are routinely conducted in shake flasks. In contrast to instrumented bioreactors, reliable options for non-invasive, time-resolved monitoring of the culture status in shake flasks are lacking. The Respiration Activity Monitoring Respiration Activity Monitoring System system was used to determine the oxygen transfer rate (OTR) in shake flasks. It was proven that the OTR could be regarded as equal to the oxygen uptake rate as the change of the dissolved oxygen concentration in the liquid phase over time was negligibly small. Thus, monitoring the oxygen transfer rate (OTR) was used to increase the information content from shake flask experiments. The OTR of a Chinese hamster ovary cell line was monitored by applying electrochemical sensors. Glass flasks stoppered with cotton plugs and polycarbonate flasks stoppered with vent-caps were compared in terms of mass transfer characteristics and culture behavior. Similar mass transfer resistances were determined for both sterile closures. The OTR was found to be well reproducible within one experiment (standard deviation <10%). It correlated with changes in cell viability and depletion of carbon sources, thus, giving more profound insights into the cultivation process. Culture behavior in glass and polycarbonate flasks was identical. Monitoring of the OTR was applied to a second culture medium. Media differed in the maximum OTR reached during cultivation and in the time when all carbon sources were depleted. By applying non-invasive, parallelized, time-resolved monitoring of the OTR, the information content and amount of data from shake flask experiments was significantly increased compared to manual sampling and offline analysis. The potential of the technology for early-stage process development was demonstrated.


Author(s):  
Luis Peña-Ortiz ◽  
Ivan Schlembach ◽  
Gerald Lackner ◽  
Lars Regestein

Mycofactocin (MFT) is a recently discovered glycosylated redox cofactor, which has been associated with the detoxification of antibiotics in pathogenic mycobacteria, and, therefore, of potential medical interest. The MFT biosynthetic gene cluster is commonly found in mycobacteria, including Mycobacterium tuberculosis, the causative agent of tuberculosis. Since the MFT molecule is highly interesting for basic research and could even serve as a potential drug target, large-scale production of the molecule is highly desired. However, conventional shake flask cultivations failed to produce enough MFT for further biochemical characterization like kinetic studies and structure elucidation, and a more comprehensive study of cultivation parameters is urgently needed. Being a redox cofactor, it can be hypothesized that the oxygen transfer rate (OTR) is a critical parameter for MFT formation. Using the non-pathogenic strain Mycobacterium smegmatis mc2 155, shake flask experiments with online measurement of the oxygen uptake and the carbon dioxide formation, were conducted under different levels of oxygen supply. Using liquid chromatography and high-resolution mass spectrometry, a 4–8 times increase of MFT production was identified under oxygen-limited conditions, in both complex and mineral medium. Moreover, the level of oxygen supply modulates not only the overall MFT formation but also the length of the glycosidic chain. Finally, all results were scaled up into a 7 L stirred tank reactor to elucidate the kinetics of MFT formation. Ultimately, this study enables the production of high amounts of these redox cofactors, to perform further investigations into the role and importance of MFTs.


2008 ◽  
Vol 7 (3) ◽  
pp. 199-211 ◽  
Author(s):  
Dan Cascaval ◽  
Anca-Irina Galaction ◽  
Stefanica Camarut ◽  
Radu Z. Tudose

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