title: Enhancing Indigo Production by Over-Expression of the Styrene Monooxygenase in Pseudomonas putida.
abstract: As an important traditional blue dye, indigo has been used in food and textile industry for centuries, which can be produced via the styrene oxygenation pathway in Pseudomonas putida. Hence, the styrene monooxygenase gene styAB and oxide isomerase gene styC are over-expressed in P. putida to investigate their roles in indigo biosynthesis. RT-qPCR analysis indicated that transcriptions of styA and styB were increased by 2500- and 750-folds in the styAB over-expressed strain B4-01, compared with the wild-type strain B4, consequently significantly enhancing the indole monooxygenase activity. Transcription of styC was also increased by 100-folds in the styC over-expressed strain B4-02. Besides, styAB over-expression slightly up-regulated the transcription of styC in B4-01, while styC over-expression hardly exerted an effect on the transcriptional levels of styA and styB and indole monooxygenase activity in B4-02. Furthermore, shaking flask experiments showed that indigo production in B4-01 reached 52.13 mg L(-1) after 24 h, which was sevenfold higher than that in B4. But no obvious increase in indigo yield was observed in B4-02. Over-expression of styAB significantly enhanced the indigo production, revealing that the monooxygenase STYAB rather than oxide isomerase STYC probably acted as the key rate-limiting enzyme in the indigo biosynthesis pathway in P. putida. This work provided a new strategy for enhancing indigo production in Pseudomonas.
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, 17 Qing Hua East Road, Hai Dian District, Beijing, 100083, China.|School of Food and Chemical Engineering, Beijing Technology and Business University, 11 & 33 Fu Cheng Road, Hai Dian District, Beijing, 100048, China.|Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University, Beijing, China.|Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, No. 11, Fu Cheng Road, Hai Dian District, Beijing, 100048, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Cheng
authors.affiliation: School of Food and Chemical Engineering, Beijing Technology and Business University, 11 & 33 Fu Cheng Road, Hai Dian District, Beijing, 100048, China.|Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University, Beijing, China.|Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, No. 11, Fu Cheng Road, Hai Dian District, Beijing, 100048, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Yin
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, 17 Qing Hua East Road, Hai Dian District, Beijing, 100083, China.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
authors.affiliation: School of Food and Chemical Engineering, Beijing Technology and Business University, 11 & 33 Fu Cheng Road, Hai Dian District, Beijing, 100048, China.|Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University, Beijing, China.|Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, No. 11, Fu Cheng Road, Hai Dian District, Beijing, 100048, China.
authors.forename: Baoguo
authors.initials: B
authors.lastname: Sun
authors.affiliation: School of Food and Chemical Engineering, Beijing Technology and Business University, 11 & 33 Fu Cheng Road, Hai Dian District, Beijing, 100048, China.|Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University, Beijing, China.|Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, No. 11, Fu Cheng Road, Hai Dian District, Beijing, 100048, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Hao
authors.affiliation: School of Food and Chemical Engineering, Beijing Technology and Business University, 11 & 33 Fu Cheng Road, Hai Dian District, Beijing, 100048, China. wctbtbu@hotmail.com.|Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University, Beijing, China. wctbtbu@hotmail.com.|Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, No. 11, Fu Cheng Road, Hai Dian District, Beijing, 100048, China. wctbtbu@hotmail.com.
authors.forename: Chengtao
authors.initials: C
authors.lastname: Wang
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D015870:Gene Expression; D007211:Indoles / Q000378:metabolism*; D010105:Oxygenases / Q000235:genetics* / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D007211:Indoles; C030374:indole; D010105:Oxygenases; C026721:styrene monooxygenase