Characterization and Expression of the Structural Gene for Pullulanase, a Maltose-inducible Secreted Protein of Klebsiella Pneumoniae

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1985 Nov 1

PMID 3902791

Citations 41

Authors

S Michaelis

C Chapon

C dEnfert

A P Pugsley

M Schwartz

Affiliations

Soon will be listed here.

Abstract

Some strains of Klebsiella pneumonia secrete pullulanase, a debranching enzyme which produces linear molecules (maltodextrins, amylose) from amylopectin and glycogen. pulA, the structural gene for pullulanase, was introduced into Escherichia coli, either on a multiple-copy-number plasmid or as a single copy in the chromosome. When in E. coli, pulA was controlled by malT, the positive regulatory gene of the maltose regulon. Indeed, pulA expression was undetectable in a malT-negative mutant and constitutive in a malTc strain. Furthermore, the plasmid carrying pulA titrated the MalT protein. When produced in E. coli, pullulanase was not localized in the same way as in K. pneumoniae. In the latter case it was first exported to the outer membrane, with which it remained loosely associated, and was then released into the growth medium. In E. coli the enzyme was distributed both in the inner and the outer membranes and was never released into the growth medium.

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References

Wallenfels K, Bender H, Rached J . Pullulanase from Aerobacter aerogenes; production in a cell-bound state. Purification and properties of the enzyme. Biochem Biophys Res Commun. 1966; 22(3):254-61. DOI: 10.1016/0006-291x(66)90474-8. View

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

Eisele B, Rasched I, Wallenfels K . Molecular characterization of pullulanase from Aerobacter aerogenes. Eur J Biochem. 1972; 26(1):62-7. DOI: 10.1111/j.1432-1033.1972.tb01739.x. View

Dessein A, Schwartz M . Is there a pullulanase in escherichia coli?. Eur J Biochem. 1974; 45(2):363-6. DOI: 10.1111/j.1432-1033.1974.tb03561.x. View

Hope G, Dean A . Pullulanase synthesis in klebsiella (aerobacter) aerogenes strains growing in continuous culture. Biochem J. 1974; 144(2):403-11. PMC: 1168509. DOI: 10.1042/bj1440403. View

Casadaban M . Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976; 104(3):541-55. DOI: 10.1016/0022-2836(76)90119-4. View

Murray N, Brammar W, Murray K . Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977; 150(1):53-61. DOI: 10.1007/BF02425325. View

Hohn B, Murray K . Packaging recombinant DNA molecules into bacteriophage particles in vitro. Proc Natl Acad Sci U S A. 1977; 74(8):3259-63. PMC: 431522. DOI: 10.1073/pnas.74.8.3259. View

Wohner G, Wober G . Pullulanase, an enzyme of starch catabolism, is associated with the outer membrane of Klebsiella. Arch Microbiol. 1978; 116(3):303-10. DOI: 10.1007/BF00417856. View

10.

Debarbouille M, Shuman H, Silhavy T, Schwartz M . Dominant constitutive mutations in malT, the positive regulator gene of the maltose regulon in Escherichia coli. J Mol Biol. 1978; 124(2):359-71. DOI: 10.1016/0022-2836(78)90304-2. View

11.

Wandersman C, Schwartz M, Ferenci T . Escherichia coli mutants impaired in maltodextrin transport. J Bacteriol. 1979; 140(1):1-13. PMC: 216772. DOI: 10.1128/jb.140.1.1-13.1979. View

12.

Pugsley A, Schnaitman C . Factors affecting the electrophoretic mobility of the major outer membrane proteins of Escherichia coli in polyacrylamide gels. Biochim Biophys Acta. 1979; 581(1):163-78. DOI: 10.1016/0005-2795(79)90233-2. View

13.

Reeve J . Use of minicells for bacteriophage-directed polypeptide synthesis. Methods Enzymol. 1979; 68:493-503. DOI: 10.1016/0076-6879(79)68038-2. View

14.

Chapon C . Role of the catabolite activator protein in the maltose regulon of Escherichia coli. J Bacteriol. 1982; 150(2):722-9. PMC: 216422. DOI: 10.1128/jb.150.2.722-729.1982. View

15.

Blumenberg M, Yanofsky C . Regulatory region of the Klebsiella aerogenes tryptophan operon. J Bacteriol. 1982; 152(1):49-56. PMC: 221373. DOI: 10.1128/jb.152.1.49-56.1982. View

16.

Raibaud O, Debarbouille M, Schwartz M . Use of deletions created in vitro to map transcriptional regulatory signals in the malA region of Escherichia coli. J Mol Biol. 1983; 163(3):395-408. DOI: 10.1016/0022-2836(83)90065-7. View

17.

Bedouelle H . Mutations in the promoter regions of the malEFG and malK-lamB operons of Escherichia coli K12. J Mol Biol. 1983; 170(4):861-82. DOI: 10.1016/s0022-2836(83)80192-2. View

18.

Chapon C . Expression of malT, the regulator gene of the maltose region in Escherichia coli, is limited both at transcription and translation. EMBO J. 1982; 1(3):369-74. PMC: 553051. DOI: 10.1002/j.1460-2075.1982.tb01176.x. View

19.

Raibaud O, Mock M, Schwartz M . A technique for integrating any DNA fragment into the chromosome of Escherichia coli. Gene. 1984; 29(1-2):231-41. DOI: 10.1016/0378-1119(84)90183-5. View

20.

Raibaud O, Gutierrez C, Schwartz M . Essential and nonessential sequences in malPp, a positively controlled promoter in Escherichia coli. J Bacteriol. 1985; 161(3):1201-8. PMC: 215027. DOI: 10.1128/jb.161.3.1201-1208.1985. View