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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 8, No. 2, 2005, pp. 185-196
Bioline Code: ej05021
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 8, No. 2, 2005, pp. 185-196

 en Peroxidase production from hairy root cultures of red beet ( Beta vulgaris check for this species in other resources )
Rudrappa ,Thimmaraju; Neelwarne, Bhagyalakshmi; Kumar, Vinod; Lakshmanan, Venkatachalam; Venkataramareddy, Sreedhar Reddampalli & Aswathanarayana, Ravishankar Gokare


The genetically transformed roots of red beet have been shown, for the first time, to produce very high levels of peroxidase (POD; EC accounting for 1.21 x 106 Units L-1. Of the ten clones established using different strains of Agrobacterium rhizogenes check for this species in other resources , one was that from the strain LMG-150, three each from A 2/83, A 20/83 and A4. All the clones showed true integration of T-DNA when tested by PCR and Southern hybridization methods. Each clone differed significantly from the others in growth, hormone dependency and POD production where LMG-150 produced highest biomass (140 g FW L-1) as well as POD (ranging from 8000-9000 U g-1 FW and 1.18 x 106 U L-1 with a specific activity of 600 U mg-1 protein) on hormone-free medium, both in shake-flask as well as in bioreactor with a further enhancement to 1.21 x 106 U L-1 upon the addition of extra calcium chloride (5 mM). PAGE with active staining showed 4 distinct bands of Rm 0.06, 0.16, 0.25, 0.38 and 0.46 in the biomass and bands at Rm 0.06, 0.16, 0.25 and one extra band of Rm 0.575 in the spent medium where isozymes of Rm 0.38 and 0.46 were totally absent. The pH optima and other properties were grossly comparable with the standard horse-radish POD (HRP) with better thermal stability than HRP and therefore, the present source appears to offer a cheaper and additional alternative for the commercial production of POD.

Agrobacterium rhizogenes, auxins, elicitation, T-DNA, thermostability, transformation.

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