caa a22 4500 605498040 CHVBK 20210128100544.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6083-y doi (NATIONALLICENCE)springer-10.1007/s00253-014-6083-y Addition of methionine and low cultivation temperatures increase palmitoleic acid production by engineered Saccharomyces cerevisiae [Elektronische Daten] [Yasushi Kamisaka, Kazuyoshi Kimura, Hiroshi Uemura, Masakazu Yamaoka] Palmitoleic acid (POA) has recently gained attention for its health benefits and as a potential resource for industrial feedstock. This study focused on the use of Saccharomyces cerevisiae, which has a high POA content but low lipid content, for POA production. We created an oleaginous S. cerevisiae as a dga1 mutant overexpressing Dga1p lacking the N-terminal 29 amino acids (Dga1∆Np). This was performed to further increase POA content in the oleaginous S. cerevisiae through optimization of culture conditions and genetic modifications. We found that high concentrations of methionine (2.0g/l) increased POA production in a concentration-dependent way, while other amino acids such as cysteine, glycine, and glutamine showed no effect. It was not clear if the effect of methionine was mediated through S-adenosylmethionine, mainly because its addition did not increase POA content as did the addition of methionine. We increased POA content up to 55% by incubation of the dga1 transformant in a medium containing 2g/l methionine at lower than normal temperatures ranging from 20 to 25°C. Cultivation at such temperatures increased dry cell weight, but did not affect the lipid content, thereby increasing total POA production. The effects of methionine and low temperatures (20-25°C) on POA content were more apparent in the strains overexpressing Dga1∆Np than those harboring empty vectors, which was consistent with the observation that POA was enriched in triacylglycerol. Overexpression of Ole1p, the enzyme responsible for POA production, did not increase POA content of the dga1 mutant overexpressing Dga1∆Np, but increased that of the wild-type strain overexpressing Dga1∆Np. The results suggested that genomic Ole1p in the dga1 mutant was active enough to achieve the optimal POA production under these conditions. Finally, the POA production by the S. cerevisiae transformant was increased 2.5-fold, which demonstrates that oleaginous S. cerevisiae is a potential source of POA. Springer-Verlag Berlin Heidelberg, 2014 DGA1 nationallicence Diacylglycerol acyltransferase nationallicence Low temperature nationallicence Methionine nationallicence OLE1 nationallicence Palmitoleic acid nationallicence Saccharomyces cerevisiae nationallicence Kamisaka Yasushi Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 305-8566, Tsukuba, Ibaraki, Japan aut Kimura Kazuyoshi Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 305-8566, Tsukuba, Ibaraki, Japan aut Uemura Hiroshi Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 305-8566, Tsukuba, Ibaraki, Japan aut Yamaoka Masakazu Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 305-8566, Tsukuba, Ibaraki, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 201-210 0175-7598 99:1<201 2015 99 253 https://doi.org/10.1007/s00253-014-6083-y text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6083-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kamisaka Yasushi Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 305-8566, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Kimura Kazuyoshi Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 305-8566, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Uemura Hiroshi Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 305-8566, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Yamaoka Masakazu Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 305-8566, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 201-210 0175-7598 99:1<201 2015 99 253