Molecular Cloning and Expression of a Xylanase Gene of Alkalophilic Aeromonas sp. no. 212 in Escherichia coli

The Riken Institute, Wako-shi, Saitama-ken 351-01, Japan ABSTRACT Summary: A gene coding for a xylanase activity of alkalophilic Aeromonas sp. no. 212 (ATCC 31085) was cloned in Escherichia coli HB101 with pBR322. Plasmid pAX1 was isolated from transformants producing xylanase, and the xylanase gene was located in a 6.0 kb Hind III fragment. The pAX1-encoded xylanase activity in E. coli HB101 was about 80 times higher than that of xylanase L in alkalophilic Aeromonas sp. no. 212. About 40% of the enzyme activity was observed in the periplasmic space of E. coli HB101. The pAX1-encoded xylanase had the same enzymic properties as those of xylanase L produced by alkalophilic Aeromonas sp. no. 212, but its molecular weight was lower (135000 vs 145000, as estimated by SDS polyacrylamide gel electrophoresis).