Efficient production of levan using a recombinant yeast Saccharomyces cerevisiae hypersecreting a bacterial levansucrase

Author:

Ko Hyunjun12,Bae Jung-Hoon1,Sung Bong Hyun12,Kim Mi-Jin1,Kim Chul-Ho3,Oh Baek-Rock3,Sohn Jung-Hoon12

Affiliation:

1. grid.249967.7 0000 0004 0636 3099 Synthetic Biology and Bioengineering Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB) 125 Gwahak-ro, Yuseong-Gu 34141 Daejeon Republic of Korea

2. grid.412786.e 0000 0004 1791 8264 Department of Biosystems and Bioengineering, KRIBB School of Biotechnology Korea University of Science and Technology (UST) 217 Gajeong-ro, Yuseong-Gu 34113 Daejeon Republic of Korea

3. grid.249967.7 0000 0004 0636 3099 Microbial Biotechnology Research Center, Jeonbuk Branch Institute Korea Research Institute of Bioscience and Biotechnology (KRIBB) 181, Ipsin-gil 56212 Jeongeup-si Jeollabuk-do Republic of Korea

Abstract

Abstract Levan is a fructose polymer with diverse applications in the food and medical industries. In this study, levansucrase from Rahnella aquatilis (RaLsrA) was hyper-secreted using a Saccharomyces cerevisiae protein secretion system. An optimal secretion signal, a translation fusion partner (TFP) containing an N-terminal 98 amino acid domain from a mitochondrial inner membrane protein, UTH1, was employed to secrete approximately 50 U/mL of bioactive RaLsrA into culture media with 63% secretion efficiency by fed-batch fermentation. Although the purified RaLsrA was useful for enzymatic conversion of high-molecular-weight levan of approximately 3.75 × 106 Da, recombinant yeast secreting RaLsrA could produce levan more efficiently by microbial fermentation. In a 50-L scale fermenter, 76-g/L levan was directly converted from 191-g/L sucrose by recombinant yeast cells, attaining an 80% conversion yield and 3.17-g/L/h productivity. Thus, we developed a cost-effective and industrially applicable production system for food-grade levan.

Publisher

Oxford University Press (OUP)

Subject

Applied Microbiology and Biotechnology,Biotechnology,Bioengineering

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