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Research Project: New Sustainable Processing Technologies to Produce Healthy, Value-Added Foods from Specialty Crops

Location: Healthy Processed Foods Research

Title: Optimization of culturing conditions of recombined Escherichia coli to produce umami octopeptide-containing protein

Author
item Zhang, Yin - Sichuan University
item Wei, Xiong - Sichuan University
item Lu, Zhou - Sichuan University
item Pan, Zhongli
item Gou, Xinhua - Sichuan University
item Venkitasamy, Chandrasekar - University Of California

Submitted to: Journal of Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/17/2017
Publication Date: 1/19/2017
Citation: Zhang, Y., Wei, X., Lu, Z., Pan, Z., Gou, X., Venkitasamy, C. 2017. Optimization of culturing conditions of recombined Escherichia coli to produce umami octopeptide-containing protein. Journal of Food Chemistry. 227:78–84. doi:10.1016/j.foodchem.2017.01.096.

Interpretive Summary: Umami substances are very important for food seasoning and healthy eating. There are 52 reported peptides showing umami taste in recent years, and twenty of these peptides are questioned for existence of umami taste. The most possible reason leading to the disputation of the taste of umami peptide might be the use of synthesized peptide to verify the taste of natural peptide. Therefore, it is important to develop methods to prepare natural peptide which can be used verify the umami taste. In this study, we used gene engineering tool and recombinant expression system to structure the Escherichia coli. The results of SDS-PAGE of the recombinant protein indicated that the recombinant expression system was successfully structured. The optimal culturing process conditions predicted with the fitting model were OD600 value of 35 0.5, IPTG of 0.6mM, culturing temperature of 28.75 ºC and culturing time of 5h. The method of preparation of natural umami peptide described in this study can be easily applied for the industrial production of LGAGGSLA.

Technical Abstract: Umami peptide is a recently found umami ingredient and its taste is widely disputed. The most possible reason leading to the disputation of the taste of umami peptide might be the use of synthesized peptide to verify the taste of natural peptide. In this research, a novel method was developed to prepare the natural peptide which could be useful to verify the taste of umami peptide. A controversial octopeptide was selected and the gene engineering tool was used to structure its Escherichia coli. expressing vector. Response surface method was adopted to optimize the expression conditions of the recombinant protein. The results of SDS-PAGE of the recombinant protein indicated that the recombinant expression system was successfully structured. The fitting results of the response surface experiment showed that the OD600 value was the key factor which influenced the expression of the recombinant protein. The optimal culturing process conditions predicted with the fitting model were OD600 value of 35 0.5, IPTG of 0.6mM, culturing temperature of 28.75 ºC and culturing time of 5h.