Production of Limonoids with Insect Antifeedant Activity in a Two-Stage Bioreactor Process with Cell Suspension Culture of Azadirachta indica

Appl Biochem Biotechnol (2015) 177:334345 DOI 10.1007/s12010-015-1745-5

Andrs Vsquez-Rivera1 & Diego Chicaiza-Finley1 & Rodrigo A. Hoyos1 & Fernando Orozco-Snchez1

Received: 27 January 2015 /Accepted: 6 July 2015 / Published online: 4 August 2015# Springer Science+Business Media New York 2015

Abstract Neem tree (Azadirachta indica) cell suspension culture is an alternative for the production of limonoids for insect control that overcomes limitations related to the supply of neem seeds. To establish conditions for cell growth and azadiracthin-related limonoid production, the effect of different sucrose concentrations, nitrate and phosphate in Murashige and Skoog (MS) medium, and the addition of one precursor and three elicitors was evaluated in shake flasks. The process was scaled up to a 3-l stirred tank bioreactor in one- and two-stage batch cultivation. In shake flasks, more than fivefold increase in the production of limonoids with the modified MS medium was observed (increase from 0.77 to 4.52 mg limonoids/g dry cell weight, DCW), while an increase of more than fourfold was achieved by adding the elicitors chitosan, salicylic acid, and jasmonic acid together (increase from 1.03 to4.32 mg limonoids/g DCW). In the bioreactor, the volumetric production of limonoids was increased more than threefold with a two-stage culture in day 18 (13.82 mg limonoids/l in control single-stage process and 41.44 mg/l in two-stage process). The cultivation and operating mode of the bioreactor reported in this study may be adapted and used in optimization and process plant development for production of insect antifeedant limonoids with A. indica cell suspension cultures.

* Fernando Orozco-Snchez [email protected]

Andrs Vsquez-Rivera [email protected]

Diego Chicaiza-Finley [email protected]

Rodrigo A. Hoyos [email protected]

1 Grupos de Biotecnologa Industrial y Biotecnologa Vegetal, Facultad de Ciencias, Universidad

Nacional de Colombia Sede Medelln, Calle 59A No 63- 20, Medelln, Colombia

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Web End = Production of Limonoids with Insect Antifeedant Activity in a Two-Stage Bioreactor Process with Cell Suspension Culture of Azadirachta indica

Appl Biochem Biotechnol (2015) 177:334345 335

Keywords Azadirachta indica . Plant cell culture . Bioreactor . Two-stage cultivation .

Limonoids . Antifeedant activity

Introduction

The use of chemical pesticides in agriculture has caused serious ecological and health...