期刊
BIORESOURCE TECHNOLOGY
卷 245, 期 -, 页码 899-905出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2017.07.116
关键词
Kluyveromyces marxianus; Astaxanthin-biosynthesis genes; Repeated genome integration; Astaxanthin; Site-directed mutagenesis
资金
- Intelligenomics Biotech. Co. Ltd.
- Ministry of Science and Technology, Taiwan [MOST 104-2311-B-039-001-MY3, MOST 104-2621-M-039-001, MOST 104-2621-M-005-003-MY3, MOST 105-2621-M-039-001-MY2, MOST 106-2622-8-005-003-TB1]
- Academia Sinica
- China Medical University Hospital [DMR-104-090, DMR-106-120]
In this study, an astaxanthin-biosynthesis Kluyveromyces marxianus strain Sm23 was first constructed, which could produce 31 mu g/g DCW astaxanthin. Then, repeated genome integration of the key astaxanthin biosynthesis genes Hpchyb and bkt was done to increase gene copy number and astaxanthin yield. Four improved strains were obtained and the yield of astaxanthin and the total yield of carotenoids in a strain increased with the copy numbers of Hpchyb and bkt. To improve the yield further, the gene Hpchyb from Haematococcus pluvialis was modified by site-directed mutagenesis to increase the enzyme efficiency or/and to prevent the heterologous protein degradation by ubiquitination. Using repeated-integration approach of bkt and the mutated Hpchyb into Sm23, the S3-2 strain was obtained and shown to produce the 3S, 3 ' S-astaxanthin at 9972 mu g/g DCW in a 5 L fermentor.
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