期刊
FOODS
卷 11, 期 14, 页码 -出版社
MDPI
DOI: 10.3390/foods11142117
关键词
indigo; monooxygenase; tryptophanase; molecular chaperone; Escherichia coli
资金
- National Natural Science Foundation of China [31401669]
- Joint Program of Beijing Natural Science Foundation
- Beijing Municipal Education Commission [KZ201910011014]
- Support Project of High-level Teachers in Beijing Municipal Universities [IDHT20180506]
- Talent Training Quality Construction-First Class Professional Construction [PXM2019-014213-000010]
In this study, a genetic manipulation strategy was developed to construct an indigo biosynthesis cell factory in E. coli for industrial production. The over-expression of styAB gene increased the capacity of indigo biosynthesis, while the expression of tryptophanase and chaperone had different effects on indigo yield.
Indigo is an important pigment widely used in industries of food, cosmetics, and textile. In this work, the styrene monooxygenase StyAB from Pseudomonas putida was co-expressed with the tryptophanase TnaA and the chaperone groES-groEL in Escherichia coli for indigo production. Over-expression of the gene styAB endowed the recombinant E. coli AB with the capacity of indigo biosynthesis from indole and tryptophan. Tryptophan fermentation in E. con AB generated about five times more indigo than that from indole, and the maximum 530 mg/L of indigo was obtained from 1.2 mg/mL of tryptophan. The gene TnaA was then co-expressed with styAB, and the tryptophanase activity significantly increased in the recombinant E. coli ABT. However, TnaA expression led to a decrease in the activity of StyAB and indigo yield in E. coli ABT. Furthermore, the plasmid pGro7 harboring groES-groEL was introduced into E. coli AB, which obviously promoted the activity of StyAB and accelerated indigo biosynthesis in the recombinant E. coli ABP. In addition, the maximum yield of indigo was further increased to 550 mg/L from 1.2 mg/mL of tryptophan in E. coli ABP. The genetic manipulation strategy proposed in this work could provide new insights into construction of indigo biosynthesis cell factory for industrial production.
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