Home LiteratureArticle Details
PMID: 1745226 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Molecular cloning and nucleotide sequence of the glycogen branching enzyme gene (glgB) from Bacillus stearothermophilus and expression in Escherichia coli and Bacillus subtilis.

Molecular & general genetics : MGG ·Vol. 230 ·No. 1-2 ·1991-11-00 ·Pages 136-44

Kiel JA, Boels JM, Beldman G, Venema G

Abstract

The structural gene for the Bacillus stearothermophilus glycogen branching enzyme (glgB) was cloned in Escherichia coli. Nucleotide sequence analysis revealed a 1917 nucleotide open reading frame (ORF) encoding a protein with an Mr of 74787 showing extensive similarity to other bacterial branching enzymes, but with a shorter N-terminal region. A second ORF of 951 nucleotides encoding a 36971 Da protein started upstream of the glgB gene. The N-terminus of the ORF2 gene product had similarity to the Alcaligenes eutrophus czcD gene, which is involved in cobalt-zinc-cadmium resistance. The B. stearothermophilus glgB gene was preceded by a sequence with extensive similarity to promoters recognized by Bacillus subtilis RNA polymerase containing sigma factor H (E - sigma H). The glgB promoter was utilized in B. subtilis exclusively in the stationary phase, and only transcribed at low levels in B. subtilis spoOH, indicating that sigma factor H was essential for the expression of the glgB gene in B. subtilis. In an expression vector, the B. stearothermophilus glgB gene directed the synthesis of a thermostable branching enzyme in E. coli as well as in B. subtilis, with optimal branching activity at 53 degrees C.

Related Genes
czcD glgB
MeSH Terms
1,4-alpha-Glucan Branching Enzyme/genetics Amino Acid Sequence Bacillus subtilis/genetics Base Sequence Blotting, Southern Cloning, Molecular DNA, Bacterial/genetics Escherichia coli/genetics Gene Expression Geobacillus stearothermophilus/genetics Molecular Sequence Data Nucleic Acid Hybridization Open Reading Frames Plasmids Promoter Regions, Genetic Sequence Homology, Nucleic Acid beta-Galactosidase/genetics
Chemicals
DNA, Bacterial 1,4-alpha-Glucan Branching Enzyme beta-Galactosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kiel J A
Department of Genetics, Center of Biological Sciences, The Netherlands.
Boels J M
Beldman G
Venema G
References (39)
39 references, click to expand
  1. Rapid and efficient cosmid cloning.
    Nucleic Acids Res. 1981 Jul 10;9(13):2989-98 PMID: 6269067
  2. Thermal stability and protein structure.
    Biochemistry. 1979 Dec 11;18(25):5698-703 PMID: 518863
  3. Synonymous codon usage in Bacillus subtilis reflects both translational selection and mutational biases.
    Nucleic Acids Res. 1987 Oct 12;15(19):8023-40 PMID: 3118331
  4. Multiple procaryotic ribonucleic acid polymerase sigma factors.
    Microbiol Rev. 1986 Sep;50(3):227-43 PMID: 3095618
  5. Improved tools for biological sequence comparison.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444-8 PMID: 3162770
  6. Physiology, biochemistry and genetics of bacterial glycogen synthesis.
    Adv Microb Physiol. 1989;30:183-238 PMID: 2700539
  7. A simple method for displaying the hydropathic character of a protein.
    J Mol Biol. 1982 May 5;157(1):105-32 PMID: 7108955
  8. Localization of a second SigH promoter in the Bacillus subtilis sigA operon and regulation of dnaE expression by the promoter.
    J Bacteriol. 1990 Oct;172(10):5631-6 PMID: 1698762
  9. A comprehensive sequence analysis program for the IBM personal computer.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):581-99 PMID: 6546431
  10. Bacillus subtilis requires a "stringent" Shine-Dalgarno region for gene expression.
    DNA. 1984;3(1):17-21 PMID: 6199171
  11. Bacterial glycogen synthesis and its regulation.
    Annu Rev Microbiol. 1984;38:419-58 PMID: 6093684
  12. Ultraviolet inactivation and excision-repair in Bacillus subtilis. I. Construction and characterization of a transformable eightfold auxotrophic strain and two ultraviolet-sensitive derivatives.
    Mutat Res. 1972 May;15(1):1-10 PMID: 4623569
  13. Sigma H-directed transcription of citG in Bacillus subtilis.
    J Bacteriol. 1989 Nov;171(11):5928-32 PMID: 2509422
  14. Cloning and nucleotide sequence of the highly thermostable neutral protease gene from Bacillus stearothermophilus.
    J Gen Microbiol. 1988 Jul;134(7):1883-92 PMID: 3149972
  15. Biosynthesis of bacterial glycogen. Purification and properties of the Escherichia coli b alpha-1,4,-glucan: alpha-1,4-glucan 6-glycosyltansferase.
    Biochemistry. 1977 Aug 9;16(16):3693-9 PMID: 407932
  16. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers.
    Gene. 1982 Oct;19(3):259-68 PMID: 6295879
  17. Improved estimation of secondary structure in ribonucleic acids.
    Nat New Biol. 1973 Nov 14;246(150):40-1 PMID: 4519026
  18. Nucleotide sequence and promoter region for the neutral protease gene from Bacillus stearothermophilus.
    J Bacteriol. 1985 Sep;163(3):824-31 PMID: 2993245
  19. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  20. The degree of branching in (alpha 1,4)-(alpha 1,6)-linked glucopolysaccharides is dependent on intrinsic properties of the branching enzymes.
    Eur J Biochem. 1987 Oct 15;168(2):393-7 PMID: 2959476
  21. The fine structure of the branched alpha-D-glucan from the blue-green alga Anacystis nidulans: comparison with other bacterial glycogens and phytoglycogen.
    Carbohydr Res. 1975 Feb;39(2):295-302 PMID: 806344
  22. Bacillus subtilis spo0H gene.
    J Bacteriol. 1984 Feb;157(2):405-12 PMID: 6420389
  23. Polysaccharide that may serve as a carbon and energy storage compound for sporulation in Bacillus cereus.
    J Bacteriol. 1974 Oct;120(1):399-406 PMID: 4214355
  24. The effect of restriction on shotgun cloning and plasmid stability in Bacillus subtilis Marburg.
    Mol Gen Genet. 1987 Sep;209(2):335-42 PMID: 2823077
  25. Toxicity of an overproduced foreign gene product in Escherichia coli and its use in plasmid vectors for the selection of transcription terminators.
    Gene. 1984 Feb;27(2):161-72 PMID: 6202587
  26. Cloning restriction fragments that promote expression of a gene in Bacillus subtilis.
    J Bacteriol. 1981 Jun;146(3):1162-5 PMID: 6787015
  27. Biosynthesis of bacterial glycogen. Primary structure of Escherichia coli 1,4-alpha-D-glucan:1,4-alpha-D-glucan 6-alpha-D-(1, 4-alpha-D-glucano)-transferase as deduced from the nucleotide sequence of the glg B gene.
    J Biol Chem. 1986 Jul 5;261(19):8738-43 PMID: 3013861
  28. Genetics of endospore formation in Bacillus subtilis.
    Annu Rev Genet. 1986;20:625-69 PMID: 3101583
  29. Cloning and expression of the branching enzyme gene (glgB) from the cyanobacterium Synechococcus sp. PCC7942 in Escherichia coli.
    Gene. 1989 May 15;78(1):9-17 PMID: 2527779
  30. Nucleotide sequence of the phosphoglycerate kinase gene from the extreme thermophile Thermus thermophilus. Comparison of the deduced amino acid sequence with that of the mesophilic yeast phosphoglycerate kinase.
    Biochem J. 1988 Sep 1;254(2):509-17 PMID: 3052437
  31. Biosynthesis of bacterial glycogen. Cloning of the glycogen biosynthetic enzyme structural genes of Escherichia coli.
    J Biol Chem. 1981 Jul 10;256(13):6944-52 PMID: 6263926
  32. Nucleotide sequence of the Synechococcus sp. PCC7942 branching enzyme gene (glgB): expression in Bacillus subtilis.
    Gene. 1990 Apr 30;89(1):77-84 PMID: 2142668
  33. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination.
    Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963-5 PMID: 6575390
  34. Transcription termination and the regulation of gene expression.
    Annu Rev Biochem. 1986;55:339-72 PMID: 3527045
  35. Expression and nucleotide sequence of a plasmid-determined divalent cation efflux system from Alcaligenes eutrophus.
    Proc Natl Acad Sci U S A. 1989 Oct;86(19):7351-5 PMID: 2678100
  36. A general method for the construction of Escherichia coli mutants by homologous recombination and plasmid segregation.
    Mol Gen Genet. 1987 May;207(2-3):294-301 PMID: 3112516
  37. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis.
    Gene. 1983 Dec;26(1):101-6 PMID: 6323249
  38. Structure and function of L-lactate dehydrogenases from thermophilic and mesophilic bacteria, VI. Nucleotide sequences of lactate dehydrogenase genes from the thermophilic bacteria Bacillus stearothermophilus, B. caldolyticus and B. caldotenax.
    Biol Chem Hoppe Seyler. 1987 Sep;368(9):1167-77 PMID: 3675869
  39. Glucan biosynthesis in Bacillus stearothermophilus. I. Properties of the polysaccharide.
    Arch Biochem Biophys. 1972 Mar;149(1):252-8 PMID: 5017254
Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00290661
Published
1991-11-00
Pages
136-44
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Databases
GENBANK
M35089, S55527, S55528, S55529, S55530, S55531, S55532, S63909, S69712, Z14057
External Links
PubMed source DOI: 10.1007/BF00290661
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2