GENETIC ENGINEERING OF THE ZYGOMYCETE BLAKESLEA TRISPORA FOR IMPROVED CAROTENOID BIOSYNTHESIS – TRANSFORMATION USING AGROBACTERIUM TUMEFACIENS

Ref ID: 4820

Author:

Thorsten Heinekamp1, Markus Matuschek2, André Schmidt1, Claus Bollschweiler2 and Axel A. Brakhage1

Author address:

1Institute of Microbiology, University of Hannover, Schneiderberg 50, D-30167 Hannover, Germany; Phone: +49 511 762
3819; Fax: +49 511 762 5287; Email: heinekamp@ifmb.uni-hannover.de
2BASF AG, Fine Chemicals & Biocatalysis Research, D-67056 Ludwigs

Full conference title:

7th European Conference on Fungal Genetics

Abstract:

Carotenoids fulfil essential biological functions and have important beneficial effects on human health, e.g., as antioxidants
or as potential antitumor agents. They are used commercially as food colours and animal feed supplements. To
date, carotenoids are mainly produced by chemical synthesis, but due to increasing preference of natural products, the
extraction of carotenoids from natural sources is becomimg an interesting alternative. The zygomycete Blakeslea trispora
is used as natural source for beta-carotene production. A major drawback of B. trispora is the lack of a transformation
system until now which allowed both to increase the production of beta-carotene and to produce interesting derivatives.
In general, stable transformation of zygomycetes was rarely described and appears to be difficult due to unknown biological
reasons. Consistently, all efforts to transform B. trispora via electroporation or protoplast formation with exogenous
DNA were not successful. Here, we report the transformation of B. trispora using the gram-negative bacterium
Agrobacterium tumefaciens. T-DNA mediated gene transfer in B. trispora via A. tumefaciens strain LBA4404 was achieved
by placing the Escherichia coli hygromycin resistance gene (hygromycin B phosphotransferase gene hph) under control
of the Aspergillus nidulans gpdA promoter. A. tumefaciens was able to introduce DNA into intact fungal cells such as
hyphae and germinating spores which circumvents protoplast generation. In an additional approach the B. trispora tef1
promoter was used to control the expression of the crtZ gene of the green algae Haematococcus pluvealis leading to production
of zeaxhantin in transgenic B. trispora cultures.

Abstract Number: VIIp-27

Conference Year: 2004

Link to conference website: NULL

New link: NULL

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