Background:　Current　commercial　production　of　isomalto-oligosaccharides　(IMOs)　commonly　involves　a　lengthy　multistage　process　with　low　yields.　Results:　To　improve　the　process　efficiency　for　production　of　IMOs,　we　developed　a　simple　and　efficient　method　by　using　enzyme　cocktails　composed　of　the　recombinant　Bacillus　naganoensis　pullulanase　produced　by　Bacillus　licheniformis,　alpha-amylase　from　Bacillus　amyloliquefaciens,　barley　bran　beta-amylase,　and　alpha-transglucosidase　from　Aspergillus　niger　to　perform　simultaneous　saccharification　and　transglycosylation　to　process　the　liquefied　starch.　After　13　h　of　reacting　time,　49.09%　IMOs　(calculated　from　the　total　amount　of　isomaltose,　isomaltotriose,　and　panose)　were　produced.　Conclusions:　Our　method　of　using　an　enzyme　cocktail　for　the　efficient　production　of　IMOs　offers　an　attractive　alternative　to　the　process　presently　in　use.　(C)　2017　Pontificia　Universidad　Cat\u00f3lica　de　Valpara\u00edso.　Production　and　hosting　by　Elsevier　B.V.　All　rights　reserved.