Establishment of in vitro plants, cell and tissue cultures from Oldenlandia affinis for the production of cyclic peptides

Abstract

In vitro cultured plants from Oldenlandia affinis were established from seeds and grown on a hormone-free medium. In vitro plants produced the cyclic peptide kalata B1 in concentrations of 0.67 mg g )1 dry weight after growth of 30 days. This was approximately 50% of the concentration analysed in green house plants (shoot tips), where different concentrations have been determined in leaves (1.82 mg g )1 ), shoot tips (1.36 mg g )1 ), stems (0.36 mg g )1 ), and in flowers (0.16 mg g )1 ). Callus and cell suspension cultures could be initiated from aseptic root, stem and leaf explants of O. affinis seedlings and plants. Different light intensities were shown to affect culture growth as well as chlorophyll synthesis. The friable callus was then used for the establishment of a cell suspension culture. Fresh and dry weight measurements showed that growth was optimal on MS medium supplemented with 0.4 mg l )1 2,4-dichlorophenoxyacetic acid (2,4-D). Leaf suspensions cultured on this medium showed a 4-fold increase of biomass by the first week of incubation. No quantifiable amounts of kalata B1 were produced under these conditions. Morphological differentiation seems to be essential for cyclic peptide production. Therefore, several undifferentiated as well as organised cell lines of O. affinis have been developed. These cell lines will constitute a worthwhile starting point for the optimisation of kalata B1 synthesis in liquid media to the objective of producing cyclic peptides under controlled and defined conditions in bioreactors.

In-vivo Hepatoprotective Activity of Methanolic Extracts of Sphaeranthus amaranthoides and Oldenlandia umbellate

“…The genus O. umbellate (family Rubiaceae) consists of different species, many of which are used in traditional medicine. 6 O. umbellate is commonly known as Indian madder, known to yield a color-fast red dye from its roots and has been used in diverse applications since ancient times. The root bark, preferably of a 2 year old plant, when used with a mordant will confer red color to calico, wool, and silk fabrics.…”

Targeting Colon Drug Delivery by Natural Products

“…Being gene products, cell based production systems seem promising, but although cyclotide producing plant cell cultures have been established [63], solid phase peptide synthesis is still the method of choice [64,65].Our knowledge about their biosynthesis is yet scarce. We know the structural arrangement of the cyclotide precur-sor from cDNA, and that an asparaginyl endo-peptidase has a likely role for cleaving at the N terminal side of the mature peptide and that protein disulfide isomerases seem to play a role for their successful folding.…”

Modern pharmacognosy: Connecting biology and chemistry

“…Given their extraordinary stability and their ability to carry highly variable stretches of sequences within this stable framework, cyclotides have also become a prime target as a platform for protein engineering [21,42]. What is important for these applications are the possibilities for both chemical synthesis and cell-based production systems that have recently been reported [43,44].…”