Abstract:
PURPOSE OF WORK:The purpose of this study was to determine if Arabidopsis protoplast transfection could be scaled up, from the commonly used cell-based studies, to be used in triterpenoid production assays as an in planta alternative/complement to other expression systems. Enzyme activities are often identified using heterologous expression systems such as yeast cells. These systems, however, may be incompatible for expressing enzymes involved in specialized (secondary) metabolism. Previous reports with long-term in planta expression systems show that the activity of the triterpenoid pathway can be enhanced by expressing enzymes catalyzing initial steps in the pathway. Here we show that triterpenoid production can also be enhanced in Arabidopsis mesophyll protoplasts after transfection. This system is designed to quantify changes in productivity of a plant metabolic pathway within 48 h and, as proof of concept, we show a significantly increased production of a triterpenoid by transiently expressing squalene synthase 1 (SQS1) from 0.5 pg/protoplast in mock-transfected protoplasts to 2.7 pg/protoplast in constitutively expressing SQS1 protoplasts.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Johnson EE,Jetter R,Wasteneys Gdoi
10.1007/s10529-013-1427-8subject
Has Abstractpub_date
2014-04-01 00:00:00pages
855-8issue
4eissn
0141-5492issn
1573-6776journal_volume
36pub_type
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