Cells Density Affects Cell Production of Citrus limonia in Flask and Air-Lift Bioreactor Cultures and Limonin Farming
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Date
2022-01-19
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Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract
Abstract. Japansche citroen (JC) is popular citrus rootstock in Indonesia due to well-adapted to biotic and abioticstresses. JC is naturally rich of secondary metabolites. Therefore, cells production of JC through tissue culture is potential for continuous metabolites farming in local pharmaceutical industries. The objectives of study were to to determine the best initial cells density (ICD) in flask and air-lift bioreactor (ALB) cultures for mass cells proliferation and to determine the limonin content in cells. Callus was induced from nucellus and habituated callus were collected to initiate cell suspensions. Selected cells were transferred to flask and ALB cultures at the density of 2, 4, and 6 mg/ml for 90 days of culturing. An increasing ICD affected cell growth in flask and ALB cultures. Cells in flask culture grew slower and were produced lower in quantity than in ALB. At 4 mg/ml, growth rate (GR) of cells in flask culture at day 30 reached maximum of 4.12 mg/day, 87-folds lower than in ALB that reached 360 mg/day. Limonin at 120.725 μg/g dried cells (DW) were harvested from ALB within 30 days, while total phenolic was 9.883±0.471 mg GAE/g DW and total flavonoid was 0.144±0.011 mg QE/g DW. This study confirmed the advantages of cell culture as a perpetual source for production of biomass content metabolites that amenable lower operational cost compared to natural production for local industries.
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Citrus limonia, abiotic, tissue culture