[Cloning and expression of endoglucanase of marine cold-adapted bacteria Pseudoalteromonas sp. MB-1].
The cold-adapted gram-negative rod bacterium MB-1 which could secret cellulase was screened from mud of the bottom of the Huanghai. According to the sequence of 16S rDNA, this bacterium screened was identified as one species of Pseudoalteromonas and was named as Pseudoalteromonas sp. MB-1. The gene celA encoding cold-adapted endogluanase was cloned and then jointed to pGEX-4T-1 to construct expression plasmid pGEX-celA which was expressed in E. coli BL21. Analysis to the supernatant of E. coli sonicate revealed that the concentration of GST-CelA was about 78.5 mg/L. Properties of the fusion enzyme of GST-CelA including the optimum temperature at 35 degrees C and the optimum pH about 7.2, showed that this fusion enzyme still belonged to cold-adapted enzyme and neutral enzyme. The result lays solid base for the fundamental theory and application research on cold-adapted cellulase from Pseudoalteromonas sp. MB-1.
You
Yin-wei
YW
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China. yyw30000@163.com
Wang
Tian-hong
TH
D002480:Cellulase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D003080:Cold Temperature; D004269:DNA, Bacterial / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D031161:Pseudoalteromonas / Q000145:classification* / Q000201:enzymology* / Q000235:genetics; D012336:RNA, Ribosomal, 16S; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry; D012623:Seawater / Q000382:microbiology*; D013696:Temperature
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D002480:Cellulase