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Background Microorganisms can prioritize the uptake of different sugars depending on their metabolic needs and preferences. When both D-glucose and D-xylose are present in growth media,  E. coli  cells typically consume D-glucose first and then D-xylose. Similarly, when  E. coli  BL21(DE3) is provided with both D-glucose and D-xylose under anaerobic conditions, glucose is consumed first, whereas D-xylose is consumed very slowly. Results When BL21(DE3) was adaptively evolved via subculture, the consumption rate of D-xylose increased gradually. Strains JH001 and JH019, whose D-xylose consumption rate was faster, were isolated after subculture. Genome analysis of the JH001 and JH019 strains revealed that C91A (Q31K) and C740T (A247V) missense mutations in the  xylR  gene (which encodes the XylR transcriptional activator), respectively, controlled the expression of the  xyl  operon. RT-qPCR analyses demonstrated that the XylR mutation caused a 10.9-fold and 3.5-fold increase in the expression of the  xylA  (xylose isomerase) and  xylF  (xylose transporter) genes, respectively, in the adaptively evolved JH001 and JH019 strains. A C91A adaptive mutation was introduced into a new BL21(DE3) background via single-base genome editing, resulting in immediate and efficient D-xylose consumption. Conclusions Anaerobically-adapted BL21(DE3) cells were obtained through short-term adaptive evolution and  xylR  mutations responsible for faster D-xylose consumption were identified, which may aid in the improvement of microbial fermentation technology.

Efficient anaerobic consumption of D-xylose by E. coli BL21(DE3) via xylR adaptive mutation