Heterologous expression of genes for bioconversion of xylose to xylonic acid in Corynebacterium glutamicum and optimization of the bioprocess

M S Lekshmi Sundar,Devi Rajan,Silvin Hannibal,K Madhavan Nampoothiri,Aliyath Susmitha,Volker F Wendisch,Keerthi Sasikumar

doi:10.1186/s13568-020-01003-9

Abstract

In bacterial system, direct conversion of xylose to xylonic acid is mediated through NAD-dependent xylose dehydrogenase (xylB) and xylonolactonase (xylC) genes. Heterologous expression of these genes from Caulobacter crescentus into recombinant Corynebacterium glutamicum ATCC 13032 and C. glutamicum ATCC 31831 (with an innate pentose transporter, araE) resulted in an efficient bioconversion process to produce xylonic acid from xylose. Process parameters including the design of production medium was optimized using a statistical tool, Response Surface Methodology (RSM). Maximum xylonic acid of 56.32 g/L from 60 g/L xylose, i.e. about 76.67% of the maximum theoretical yield was obtained after 120 h fermentation from pure xylose with recombinant C. glutamicum ATCC 31831 containing the plasmid pVWEx1 xylB. Under the same condition, the production with recombinant C. glutamicum ATCC 13032 (with pVWEx1 xylB) was 50.66 g/L, i.e. 69% of the theoretical yield. There was no significant improvement in production with the simultaneous expression of xylB and xylC genes together indicating xylose dehydrogenase activity as one of the rate limiting factor in the bioconversion. Finally, proof of concept experiment in utilizing biomass derived pentose sugar, xylose, for xylonic acid production was also carried out and obtained 42.94 g/L xylonic acid from 60 g/L xylose. These results promise a significant value addition for the future bio refinery programs.

Highlights

D-xylonic acid, an oxidation product of xylose, is a versatile platform chemical with multifaceted applications in the fields of food, pharmaceuticals, and agriculture
In addition to ATCC 13032 wild type, we explored the C.glutamicum ATCC 31831 culture which contains a pentose transporter gene which enables the uptake of pentose sugar (Kawaguchi et al 2009; Choi et al 2019)
To check xylonic acid production from xylose, the C. glutamicum ATCC 31831 transformants harboring pVWEx1-xylB, pVWEx1-xylC and pVWEx1-xylBC were cultivated in CGXII medium containing 5 g/L of glucose as the carbon source for initial cell growth and 35 g/L of xylose as the substrate for xylonic acid production

Summary

Introduction

D-xylonic acid, an oxidation product of xylose, is a versatile platform chemical with multifaceted applications in the fields of food, pharmaceuticals, and agriculture. It is considered by the U.S Department of Energy to be one of the 30 chemicals of highest value because it can be used in a variety of applications, including as a dispersant, pH regulator, chelator, antibiotic clarifying agent and health enhancer (Byong-Wa et al 2006; Toivari et al 2012). Biogenic production of xylonic acid has been accomplished in various microorganisms, including Escherichia coli, Saccharomyces cerevisiae and Kluyveromyces lactis by introducing xylB (encoding xylose dehydrogenase) and xylC (encoding xylonolactonase) genes from Caulobacter crescentus or Trichoderma reesei (Nygård et al 2011; Toivari et al 2012; Cao et al 2013)

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Results

Conclusion