Biocatalytic upgrading of bio-based platform chemical 5-hydroxymethylfurfural (5-HMF) to 2,5-bis(hydroxymethyl) furan (BHMF) is currently of great interest due to the product specificity, mild reaction and high efficiency. In this work, 200 mM 5-HMF could be effectively biotransformed to BHMF at 90.6% with highly 5-HMF- tolerant recombinant E. coli CCZU-K14 whole cells at pH 6.5 and 30 degrees C under the optimum reaction conditions (cosubstrate glucose 1.0 mol glucose/(mol 5-HMF), D-xylose 400 mM, L-glutamic acid 250 mM, Mg2+ 1.5 mM, 0.2 mol beta-cyclodextrin/(mol 5-HMF), CTAB (cetyltrimethyl ammonium bromide) 12.5 mM, and 0.1 g wet cells/mL). It was found that E. coli CCZU-K14 was highly tolerant to 5-HMF (up to 400 mM). Effective bioreduction of biomass-derived 5-HMF (<= 200) to BHMF was successfully demonstrated in this study. In conclusion, this strategy showed high potential application for the synthesis of BHMF.