Biotechnological process for production of beta-dipeptides from cyanophycin on a technical scale and its optimization

Sallam A, Kast A, Przybilla S, Meiswinkel T, Steinbüchel A (2009)
Applied and environmental microbiology 75(1): 29-38.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
A triphasic process was developed for the production of beta dipeptides from cyanophycin (CGP) on a large scale. Phase I comprises an optimized acid extraction method for technical isolation of CGP from biomass. It yielded highly purified CGP consisting of aspartate, arginine, and a little lysine. Phase II comprises the fermentative production of an extracellular CGPase (CphE(al)) from Pseudomonas alcaligenes strain DIP1 on a 500-liter scale in mineral salts medium, with citrate as the sole carbon source and CGP as an inductor. During optimization, it was shown that 2 g liter(-1) citrate, pH 6.5, and 37 degrees C are ideal parameters for CphE(al) production. Maximum enzyme yields were obtained after induction in the presence of 50 mg liter(-1) CGP or CGP dipeptides for 5 or 3 h, respectively. Aspartate at a concentration of 4 g liter(-1) induced CphE(al) production with only about 30% efficiency in comparison to that with CGP. CphE(al) was purified utilizing its affinity for the substrate and its specific binding to CGP. CphE(al) turned out to be a serine protease with maximum activity at 50 degrees C and at pH 7 to 8.5. Phase III comprises degradation of CGP to beta-aspartate-arginine and beta-aspartate-lysine dipeptides with a purity of over 99% (by thin-layer chromatography and high-performance liquid chromatography), employing a crude CphE(al) preparation. Optimum degradation parameters were 100 g liter(-1) CGP, 10 g liter(-1) crude CphE(al) powder, and 4 h of incubation at 50 degrees C. The overall efficiency of phase III was 91%, while 78% (wt/wt) of the used CphE(al) powder with sustained activity toward CGP was recovered. The optimized process was performed with industrial materials and equipment and is applicable to any desired scale.
Stichworte
Peptide Hydrolases/isolation & purification; Dipeptides/metabolism; Dipeptides/isolation & purification; Thin Layer; Chromatography; Affinity; Chromatography; Biotechnology/methods; Biomass; Bacterial Proteins/isolation & purification; Bacterial Proteins/metabolism; Peptide Hydrolases/metabolism; Plant Proteins/isolation & purification; Plant Proteins/metabolism; Pseudomonas alcaligenes/enzymology
Erscheinungsjahr
2009
Zeitschriftentitel
Applied and environmental microbiology
Band
75
Ausgabe
1
Seite(n)
29-38
ISSN
0099-2240
Page URI
https://pub.uni-bielefeld.de/record/2473094

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Sallam A, Kast A, Przybilla S, Meiswinkel T, Steinbüchel A. Biotechnological process for production of beta-dipeptides from cyanophycin on a technical scale and its optimization. Applied and environmental microbiology. 2009;75(1):29-38.
Sallam, A., Kast, A., Przybilla, S., Meiswinkel, T., & Steinbüchel, A. (2009). Biotechnological process for production of beta-dipeptides from cyanophycin on a technical scale and its optimization. Applied and environmental microbiology, 75(1), 29-38. doi:10.1128/AEM.01344-08
Sallam, A., Kast, A., Przybilla, S., Meiswinkel, T., and Steinbüchel, A. (2009). Biotechnological process for production of beta-dipeptides from cyanophycin on a technical scale and its optimization. Applied and environmental microbiology 75, 29-38.
Sallam, A., et al., 2009. Biotechnological process for production of beta-dipeptides from cyanophycin on a technical scale and its optimization. Applied and environmental microbiology, 75(1), p 29-38.
A. Sallam, et al., “Biotechnological process for production of beta-dipeptides from cyanophycin on a technical scale and its optimization”, Applied and environmental microbiology, vol. 75, 2009, pp. 29-38.
Sallam, A., Kast, A., Przybilla, S., Meiswinkel, T., Steinbüchel, A.: Biotechnological process for production of beta-dipeptides from cyanophycin on a technical scale and its optimization. Applied and environmental microbiology. 75, 29-38 (2009).
Sallam, Ahmed, Kast, Alene, Przybilla, Simon, Meiswinkel, Tobias, and Steinbüchel, Alexander. “Biotechnological process for production of beta-dipeptides from cyanophycin on a technical scale and its optimization”. Applied and environmental microbiology 75.1 (2009): 29-38.

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