Production of L-Theanine Using Escherichia coli Whole-Cell Overexpressing gamma-Glutamylmethylamide Synthetase with Baker's Yeast
- 주제(키워드) L-theanine , whole-cell biocatalyst , baker's yeast , ATP regeneration
- 주제(기타) Biotechnology & Applied Microbiology
- 주제(기타) Microbiology
- 설명문(일반) [Yang, Soo-Yeon; Han, Yeong-Hoon; Park, Ye-Lim; Park, Jun-Young; No, So-Young; Park, Saerom; Park, Hyung Yeon; Yang, Yung-Hun] Konkuk Univ, Dept Biol Engn, Coll Engn, Seoul 05029, South Korea; [Jeong, Daham] Konkuk Univ, Dept Biosci & Biotechnol, Seoul 05029, South Korea; [Kim, Wooseong] Ewha Womans Univ, Coll Pharm, Seoul 03760, South Korea; [Kim, Wooseong] Ewha Womans Univ, Grad Sch Pharmaceut Sci, Seoul 03760, South Korea; [Seo, Seung-Oh] Catholic Univ Korea, Dept Food Sci & Nutr, Bucheon 14662, South Korea; [Yang, Yung-Hun] Konkuk Univ, Inst Ubiquitous Informat Technol & Applicat, Seoul 05029, South Korea
- 등재 SCIE, SCOPUS, KCI등재
- OA유형 Bronze
- 발행기관 KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
- 발행년도 2020
- 총서유형 Journal
- URI http://www.dcollection.net/handler/ewha/000000168946
- 본문언어 영어
- Published As https://dx.doi.org/10.4014/jmb.1910.10044
초록/요약
L-Theanine, found in green tea leaves has been shown to positively affect immunity and relaxation in humans. There have been many attempts to produce L-theanine through enzymatic synthesis to overcome the limitations of traditional methods. Among the many genes coding for enzymes in the L-theanine biosynthesis, glutamylmethylamide synthetase (GMAS) exhibits the greatest possibility of producing large amounts of production. Thus, GMAS from Methylovorus mays No. 9 was overexpressed in several strains including vectors with different copy numbers. BW25113(DE3) cells containing the pET24ma::gmas was selected for strains. The optimal temperature, pH, and metal ion concentration were 50 degrees C, 7, and 5 mM MnCl2, respectively. Additionally, ATP was found to be an important factor for producing high concentration of L-theanine so several strains were tested during the reaction for ATP regeneration. Baker's yeast was found to decrease the demand for ATP most effectively. Addition of potassium phosphate source was demonstrated by producing 4-fold higher L-theanine. To enhance the conversion yield, GMAS was additionally overexpressed in the system. A maximum of 198 mM L-theanine was produced with 16.5 mmol/l/h productivity. The whole-cell reaction involving GMAS has greatest potential for scale-up production of L-theanine.
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