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J Microbiol Biotechnol. 2017 Feb 28;27(2):306-315. doi: 10.4014/jmb.1611.11054.

Enhanced Production of Itaconic Acid through Development of Transformed Fungal Strains of Aspergillus terreus.

Author information

1
Green Materials and Process R&BD Group, Korea Institute of Industrial Technology, Cheonan 331822, Republic of Korea.
2
Department of Chemical and Biological Engineering, Korea University, Seoul 136713, Republic of Korea.
3
Department of Green Process and System Engineering, Korea University of Science and Technology, Daejeon 34141, Republic of Korea.
4
College of Biomedical Science, Kangwon National University, Chuncheon 24341, Republic of Korea.

Abstract

Metabolic engineering with a high-yielding mutant, A. terreus AN37, was performed to enhance the production of itaconic acid (IA). Reportedly, the gene cluster for IA biosynthesis is composed of four genes: reg (regulator), mtt (mitochondrial transporter), cad (cis-aconitate decarboxylase), and mfs (membrane transporter). By overexpressing each gene of the IA gene cluster in A. terreus AN37 transformed by the restriction enzyme-mediated integration method, several transformants showing high productivity of IA were successfully obtained. One of the AN37/cad transformants could produce a very high amount of IA (75 g/l) in shake-flask cultivations, showing an average of 5% higher IA titer compared with the high-yielding control strain. Notably, in the case of the mfs transformants, a maximal increase of 18.3% in IA production was observed relative to the control strain under the identical fermentation conditions. Meanwhile, the overexpression of reg and mtt genes showed no significant improvements in IA production. In summary, the overexpressed cis-aconitate decarboxylase (CAD) and putative membrane transporter (MFS) appeared to have positive influences on the enhanced IA productivity of the respective transformant. The maximal increases of 13.6~18.3% in IA productivity of the transformed strains should be noted, since the parallel mother strain used in this study is indeed a very high-performance mutant that has been obtained through intensive rational screening programs in our laboratory.

KEYWORDS:

Aspergillus terreus; cis-aconitate decarboxylase (CAD); itaconate gene cluster; plasma membrane transporter (MFS); protoplast transformation; restriction enzyme-mediated integration (REMI)

PMID:
27974733
DOI:
10.4014/jmb.1611.11054
[Indexed for MEDLINE]
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