Low malt beers have high sales volumes in Japan, but improving their mouthfeel, including softness, smoothness and decreasing astringency, is challenging because the compounds responsible remain unclear. In this study, beer was fractionated by preparative size-exclusion chromatography, with the polypeptide and maltodextrin fractions purified using solid-phase extraction and ion-exchange resin. Sensory data from a spike test showed that the mouthfeel (softness, smoothness, and reduced astringency) of low malt beer was improved both by the degree of polymerisation (DP) of maltodextrins (DP of 2-10; at increased concentration of 40 to 60%; P < 0.01) and by 10 - 20 kilodalton (kDa) high molecular weight (HMW) polypeptide and 2-3 kDa low molecular weight polypeptide fractions (at a 50% increase in concentration; P < 0.01). Furthermore, highly purified 10 to 20 kDa HMW polypeptides improved the softness and smoothness and decreased the astringency (at a 25% increase in concentration). This report is the first to provide experimental sensory data indicating that HMW polypeptides improve the mouthfeel of beer. Based on these findings, a new low malt beer was developed that showed significantly higher levels of the 10-20 kDa HMW polypeptides with an overall improved mouthfeel. Mass spectrometric analysis of the 10 to 20 kDa proteins identified several unique foam positive proteins, including barley dimeric alpha-amylase inhibitor-1 and non-specific lipid-transfer protein 1. These 10-20 kDa HMW proteins are likely to be responsible for the improved mouthfeel of beer. © 2020 Kirin Holdings Kabushik Kaisha Co. Ltd. Journal of the Institute of Brewing published by John Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling