An　agar-degrading　bacterium,　strain　BN3,　was　isolated　from　a　coastal　soil　sample　collected　in　Taiwan　Strait,　China　and　identified　to　be　a　novel　species　of　the　genus　Microbulbifer.　The　gene　(N3-1)　encoding　for　a　novel　β-agarase　from　the　isolate　was　cloned　and　sequenced.　It　encoded　a　mature　protein　with　274　amino　acids　and　a　calculated　molecular　mass　of　34.3　kDa.　The　deduced　amino　acid　sequence　manifested　sequence　similarity　(61–84%　identity)　to　characterized　β-agarases　in　the　glycoside　hydrolase　family　16.　The　recombinant　agarase　was　hyper-produced　extracellularly　using　Pichia　pastoris　as　the　host.　After　induction　in　a　shake　flask　for　96　h,　the　yield　of　recombinant　N3–1　protein　reached　0.406　mg/mL,　and　the　enzyme　activity　attained　502.1　U/mL.　The　enzyme　purified　by　ion　exchange　chromatography　displayed　a　specific　activity　of　6447　U/mg　at　pH　6.0　and　50　°C.　The　optimal　pH　and　temperature　for　agarase　activity　were　approximately　6　and　50　°C,　respectively.　The　pattern　of　agarose　hydrolysis　showed　that　the　enzyme　was　an　endo-type　β-agarase,　capable　of　hydrolyzing　agarose　and　Gracilaria　lemaneiformis,　with　neoagarobiose　and　neoagarotetraose　as　the　final　main　products.　©　2018