Construction of a glucaric acid biosensor for screening myo-inositol oxygenase variants.

Author: Cui WANG ¹ ; Ye LIU ¹ ; Xu GONG ¹ ; Long LIU ¹ ; Zhen KANG ¹
Author Information

1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
Publication Type:Journal Article
Keywords: Myo-inositol oxygenase; biosensor; glucaric acid; high-throughput screening
MeSH: Biosensing Techniques; Biosynthetic Pathways; Escherichia coli; Glucaric Acid; chemistry; Inositol Oxygenase; chemistry
From: Chinese Journal of Biotechnology 2018;34(11):1772-1783
CountryChina
Language:Chinese
Abstract: Glucaric acid (GA), a top value-added chemical from biomass, has been widely used for prevention and control of diseases and the production of polymer materials. In GA biosynthesis pathway, the conversion of inositol to glucuronic acid that catalyzed by myo-inositol oxygenase is the limiting step. It is necessary to improve MIOX activity. In the present study, we constructed a high-throughput screening system through combing the concentration of GA with the green fluorescent protein fluorescence intensity. By applying this screening system, three positive variants (K59V/R60A, R171S and D276A) screened from the mutant library. In comparison, the recombinant strain Escherichia coli BL21(DE3)/MU-R171S accumulated more GA, 136.5% of that of the parent strain.