Abstract: [Objective] This project aims to have a preliminary identification of the two thermotolerant fungi MHS7 and MHS9 isolated from a hot gas spring in Manasi County of Xining and study their enzyme activities.[Method]Thermotolerant fungi were isolated and screened by using a agar dilution method, and the primary identification was studied according to cultural characteristics and rDNA ITS (Internal Transcribed Spacer) sequences analysis.The amylase, protease, cellulose, lipase, xylanase and glycosidase were detected on six special selective media for thermotolerant fungus strains with the enzyme activity.[Result]Two strains of thermotolerant fungi named MHS7 and MHS9 were isolated.ITS sequence analysis identified that the strain MHS7 belonged to Gymnascella sp., and the strain MHS9 was Eurotium sp..Studies on enzyme-producing activities of strains MHS7 and MHS9 indicated that both of them produced amylase, protease, cellulose, and lipase.Of the two, the strain MHS9 had higher levels of amylase and protease activities than the strain MHS7.[Conclusion]The thermotolerant fungi MHS7 and MHS9 had complex enzyme activities and might have potential application value.