The cloned β-mannanase gene from alkalophilic Bacillus sp. AM-001 produces two β-mannanases in Escherichia coli

The gene encoding beta -mannanase was cloned from alkalophilic Bacillus sp. AM-001 into Escherichia coli JM101 by inserting HindIII-generated DNA fragments into the HindIII site of pUC19. A 2.0 kb XbaI-PstI fragment of the donor strain DNA was sufficient for beta -mannanase synthesis. The amount of...

Full description

Saved in:
Bibliographic Details
Published in:Archives of microbiology 1989-06, Vol.152 (1), p.10-15
Main Authors: AKINO, T, KATO, C, HORIKOSHI, K
Format: Article
Language:English
Subjects:
Bacillus
Bacteriology
beta -mannanase A
beta -mannanase B
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
genes
Metabolism. Enzymes
Microbiology
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The gene encoding beta -mannanase was cloned from alkalophilic Bacillus sp. AM-001 into Escherichia coli JM101 by inserting HindIII-generated DNA fragments into the HindIII site of pUC19. A 2.0 kb XbaI-PstI fragment of the donor strain DNA was sufficient for beta -mannanase synthesis. The amount of beta -mannanase expressed in E. coli JM101 harboring pMAH3 (containing a 2.4 kb XbaI-HindIII fragment) was about 24% of the activity produced by the donor strain. E. coli JM101 harboring pMAH3 was found to produce two enzymatically active beta -mannanases (A and B). These two beta -mannanases were purified to electrophoretically homogenous states. The beta -mannanase A had enzymatic properties similar to those of the beta -mannanases M-I and M-II produced by alkalophilic Bacillus sp. AM-001, and the beta mannanase B resembled its beta -mannanase M-III. In contrast to beta -mannanase production in the donor strain, that in E. coli was not inducible.
ISSN:0302-8933
1432-072X
DOI:10.1007/BF00447004