CsCCD2 Access Tunnel Design for a Broader Substrate Profile in Crocetin Production
文献类型：期刊论文
| 作者 | Liang, Nan1,2,3; Yao, Ming-Dong1,2,3; Wang, Ying1,2,3; Liu, Jia4 ; Feng, Lu4; Wang, Zhi-Ming5; Li, Xiang-Yu5; Xiao, Wen-Hai1,2,3,6; Yuan, Ying-Jin1,2,3
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| 刊名 | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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| 出版日期 | 2021-10-06 |
| 卷号 | 69期号:39页码:11626-11636 |
| ISSN号 | 0021-8561 |
| 关键词 | crocetin CCD2 access tunnel engineering Saccharomyces cerevisiae |
| DOI | 10.1021/acs.jafc.1c04588 |
| 通讯作者 | Xiao, Wen-Hai(wenhai.xiao@tju.edu.cn) |
| 英文摘要 | Crocetin, a high-value apocarotenoid in saffron, is widely applied to the fields of food and medicine. However, the existing method of obtaining crocetin through large-scale cultivation is far from meeting the market demand. Microbial synthesis of crocetin is a potential alternative to traditional resources, and it is found that carotenoid cleavage dioxygenase (CCD) is the critical enzyme to synthesize crocetin. So, in this study, we used "hybrid-tunnel" engineering to obtain variants of Crocus sativus-derived CsCCD2, essential for zeaxanthin conversion into crocetin, with a broader substrate specificity and higher catalytic efficiency. Variants including S323A, with a lower charge bias and a larger tunnel size than the wild-type, showed a 5-fold higher crocetin titer in yeast-based fermentations. S323A could also convert the beta-carotene substrate to crocetin dialdehyde and exhibited a 12.83-fold greater catalytic efficiency (k(cat)/K-m) toward zeaxanthin than the wild-type in vitro. This strategy enabled the production of 107 mg/L crocetin in 5 L fed-batch fermentation, higher than that previously reported. Our findings demonstrate that engineering access tunnels to expand the substrate profile by in silico protein design represents a viable strategy to refine the catalytic properties of enzymes across a range of applications. |
| WOS关键词 | CROCUS-SATIVUS L. ; CAROTENOID CLEAVAGE ; SAFFRON ; BIOSYNTHESIS ; CATALYZES ; PATHWAY |
| 资助项目 | National Key Research and Development Program of China[2019YFA0706900] ; National Natural Science Foundation of China[21621004] ; Key-Area Research and Development Program of Guangdong Province[2020B0303070002] |
| WOS研究方向 | Agriculture ; Chemistry ; Food Science & Technology |
| 语种 | 英语 |
| 出版者 | AMER CHEMICAL SOC |
| WOS记录号 | WOS:000706191500009 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/298478]  |
| 专题 | 中国科学院上海药物研究所 |
| 通讯作者 | Xiao, Wen-Hai |
| 作者单位 | 1.Tianjin Univ, Sch Chem Engn & Technol, Frontier Sci Ctr Synthet Biol, Tianjin 300072, Peoples R China 2.Tianjin Univ, Sch Chem Engn & Technol, Minist Educ, Key Lab Syst Bioengn, Tianjin 300072, Peoples R China 3.Tianjin Univ, Collaborat Innovat Ctr Chem Sci & Engn Tianjin, Tianjin 300072, Peoples R China 4.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 5.CABIO Biotech Wuhan Co Ltd, Wuhan 436070, Peoples R China 6.Tianjin Univ, Georgia Tech Shenzhen Inst, Shenzhen 518071, Peoples R China |
| 推荐引用方式 GB/T 7714 | Liang, Nan,Yao, Ming-Dong,Wang, Ying,et al. CsCCD2 Access Tunnel Design for a Broader Substrate Profile in Crocetin Production[J]. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY,2021,69(39):11626-11636. |
| APA | Liang, Nan.,Yao, Ming-Dong.,Wang, Ying.,Liu, Jia.,Feng, Lu.,...&Yuan, Ying-Jin.(2021).CsCCD2 Access Tunnel Design for a Broader Substrate Profile in Crocetin Production.JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY,69(39),11626-11636. |
| MLA | Liang, Nan,et al."CsCCD2 Access Tunnel Design for a Broader Substrate Profile in Crocetin Production".JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 69.39(2021):11626-11636. |
入库方式： OAI收割
来源：上海药物研究所
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