Xylitol　is　a　salutary　sugar　substitute　that　has　been　widely　used　in　the　food,　pharmaceutical,　and　chemical　industries.　Co-fermentation　of　xylose　and　glucose　by　metabolically　engineered　cell　factories　is　a　promising　alternative　to　chemical　hydrogenation　of　xylose　for　commercial　production　of　xylitol.　Here,　we　engineered　a　mutant　of　SecY　protein-translocation　channel　(SecY　[ΔP])　in　xylitol-producing　Escherichia　coli　JM109　(DE3)　as　a　passageway　for　xylose　uptake.　It　was　found　that　SecY　(ΔP)　channel　could　rapidly　transport　xylose　without　being　interfered　by　XylB-catalyzed　synthesis　of　xylitol-phosphate,　which　is　impossible　for　native　XylFGH　and　XylE　transporters.　More　importantly,　with　the　coaction　of　SecY　(ΔP)　channel　and　carbon　catabolite　repression　(CCR),　the　flux　of　xylose　to　the　pentose　phosphate　(PP)　pathway　and　the　xylitol　synthesis　pathway　in　E.　coli　could　be　automatically　controlled　in　response　to　glucose,　thereby　ensuring　that　the　mutant　cells　were　able　to　fully　utilize　sugars　with　high　xylitol　yields.　The　E.　coli　cell　factory　developed　in　this　study　has　been　proven　to　be　applicable　to　a　broad　range　of　xylose-glucose　mixtures,　which　is　conducive　to　simplifying　the　mixed-sugar　fermentation　process　for　efficient　and　economical　production　of　xylitol.　©　2021　Wiley　Periodicals　LLC