Abstract

In the industrial production of xanthan gum using Xanthomonas campestris CGMCC15155, large amounts of ethanol are required to extract xanthan gum from the fermentation broth and remove xanthomonadin impurities. To reduce the amount of ethanol and the overall production cost of xanthan gum, a xanthomonadin‐deficient strain of CGMCC15155 was constructed by inserting the Vitreoscilla globin (vgb) gene, under the control of the LacZ promoter, into the region of the pigA gene, which is involved in xanthomonadin synthesis. The insertion of vgb inactivated pigA, resulting in the production of white xanthan gum. The lack of xanthomonadins resulted in a decreased yield of xanthan gum. However, the expression product of vgb gene, VHb, could increase the metabolism of X. campestris, which allowed the production of xanthan gum to reach wild‐type levels in the engineered strain. The yield, molecular weight, and rheological properties of the xanthan gum synthesized by the engineered and wild‐type bacteria were essentially the same. When the same volume of ethanol was used, the whiteness values of the xanthan gum extracted from engineered and wild‐type bacteria were 65.20 and 38.17, respectively. To extract xanthan gum with the same whiteness, three and seven times the fermentation volume of ethanol was required for the engineered and wild‐type strains, respectively. Thus, the engineered train reduced the requirement for ethanol in xanthan gum production by 133.3%. The results demonstrated that the engineered bacteria used less ethanol, thus reducing the downstream processing cost in xanthan gum production.

Highlights

  • Polysaccharides are very important biological materials in food, medicine, and other industries

  • Recombinant plasmid pLO3-­pigA was transformed into X. campestris CGMCC15155 (XC) via conjugation between E. coli S17-­1 pLO3-­pigA and XC, which resulted in the pigA gene being knocked out by homologous recombination

  • 3.1 | Identification of genes related to xanthomonadins in Xanthomonas campestris CGMCC15155

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Summary

| INTRODUCTION

Polysaccharides are very important biological materials in food, medicine, and other industries. The production cost of food grade xanthan gum in the downstream purification steps can be as high as 50%, which would not be necessary for nonfood applications (Palaniraj & Jayaraman, 2011). One of the main problems in industrial xanthan gum production is the difficulty in stirring and ventilating the broth during fermentation, which affects oxygen transmission (Palaniraj & Jayaraman, 2011). We hypothesized that introducing the Vitreoscilla globin (vgb) gene into X. campestris might reduce the oxygen shortage, resulting from the excessive viscosity of the fermentation broth, thereby increasing the yield of xanthan gum. The expression of VHb increased the metabolism of X. campestris, allowing the engineered strain to produce xanthan gum at wild-­type levels. The results demonstrated that the engineered bacteria required less ethanol for downstream processing, reducing the cost of xanthan gum production

| MATERIALS AND METHODS
| Analytical methods
| Evaluation of rheological behavior
| RESULTS AND DISCUSSION
| CONCLUSIONS
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