A　novel　immobilized　cellulase　based　on　hydrogel　microsphere　was　established　to　accommodate　ionic　liquid　(IL)-involved　in　situ　biomass　saccharification.　For　immobilization,　poly(N-isopropylacrylamide)　(PNIPAM)　was　employed　to　immobilize　cellulase,　with　immobilization　yielded　up　to　95.6%.　Besides,　1-ethyl-3-methyllimidazolium　acetate　([EMIM]OAc)　was　found　as　an　appropriate　IL　for　in　situ　saccharification　due　to　its　relative　enzymatic　activity,　which　was　as　high　as　167.5%.　With　utilization　of　immobilized　cellulase　in　IL-pretreated　in　situ　bagasse　(BC)　saccharification,　relative　enzymatic　activity　was　40.9%,　which　was　higher　than　free　cellulase　under　5%　(v/v)　IL,　and　achieved　12.3%,　which　was　found　to　be　higher　than　free　one　in　25%　(v/v)　IL.　Moreover,　for　estimating　reusability,　relative　enzymatic　activity　on　the　6th　cycle　retained　over　85.2%.　This　novel　immobilization　system　possessed　a　remarkable　preservation　to　cellulase　in　hydrolysis　contained　interferences,　e.g.　IL,　suggesting　a　noticeable　practical　potential　in　hydrolysis　of　cost-effective　sustainable　biomass　materials.　©　2019　Elsevier　Ltd