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Escherichia coli K-12 substr. MG1655 Enzyme: endo-1,4-D-glucanase



Gene: bcsZ Accession Numbers: EG12258 (EcoCyc), b3531, ECK3516

Synonyms: bcsC, yhjM

Regulation Summary Diagram: ?

Summary:
The bcsZ gene encodes an endo-1,4-D-glucanase which belongs to glycosyl hydrolase family 8 [Park99].

BcsZ has a predicted signal sequence [Park99]; a computational prediction that BcsZ is a lipoprotein is thought to be a probable false positive [Gonnet04]. In cell fractionation assays, the enzymatic activity appeared to be primarily extracellular [Park99], although the publication reported no positive controls.

Crystal structures of apo-BcsZ and that of a catalytically inactive mutant bound to the substrate cellopentaose have been solved [Mazur11].

BcsZ is encoded in a predicted operon together with bcsA, bcsB, and bcsC. In other organisms, these genes are involved in cellulose biosynthesis, a characteristic of the rdar morphotype. However, the K-12 laboratory strain of E. coli does not show a rdar morphotype and does not produce cellulose [Zogaj01]. In E. coli K-12 strains such as W3110 and MG1655, a "domesticating SNP" consists of a point mutation that introduces a stop codon after the first 5 amino acids of the wild-type bcsQ ORF. The bcsQ ORF shown in EcoCyc begins just downstream of this SNP and appears to be non-functional. This domesticating SNP lowers expression of both bcsQ and the downstream bcsA gene. After repairing the SNP, the resulting "de-domesticated" W3110 derivative strain produces cellulose and has dramatically altered biofilm morphology [Serra13].

BcsZ: "bacterial cellulose synthesis"

Citations: [Omadjela13]

Gene Citations: [Blattner97, Sofia94, Salmon03]

Locations: extracellular space

Map Position: [3,687,178 <- 3,688,284] (79.47 centisomes)
Length: 1107 bp / 368 aa

Molecular Weight of Polypeptide: 41.7 kD (from nucleotide sequence), 40 kD (experimental) [Park99 ]

Unification Links: ASAP:ABE-0011541 , EchoBASE:EB2167 , EcoGene:EG12258 , EcoliWiki:b3531 , ModBase:P37651 , OU-Microarray:b3531 , PortEco:bcsZ , PR:PRO_000022205 , Pride:P37651 , Protein Model Portal:P37651 , RefSeq:NP_417988 , RegulonDB:EG12258 , SMR:P37651 , String:511145.b3531 , UniProt:P37651

Relationship Links: CAZy:IN-FAMILY:GH8 , InterPro:IN-FAMILY:IPR002037 , InterPro:IN-FAMILY:IPR008928 , InterPro:IN-FAMILY:IPR012341 , InterPro:IN-FAMILY:IPR019834 , PDB:Structure:3QXF , PDB:Structure:3QXQ , Pfam:IN-FAMILY:PF01270 , Prints:IN-FAMILY:PR00735 , Prosite:IN-FAMILY:PS00812

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0000272 - polysaccharide catabolic process Inferred by computational analysis [UniProtGOA11a]
GO:0005975 - carbohydrate metabolic process Inferred by computational analysis [UniProtGOA11a, GOA01]
GO:0008152 - metabolic process Inferred by computational analysis [UniProtGOA11a]
GO:0030245 - cellulose catabolic process Inferred by computational analysis [UniProtGOA11a]
Molecular Function: GO:0008810 - cellulase activity Inferred from experiment Inferred by computational analysis [GOA01a, Park99]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01]
GO:0004553 - hydrolase activity, hydrolyzing O-glycosyl compounds Inferred by computational analysis [GOA01]
GO:0016787 - hydrolase activity Inferred by computational analysis [UniProtGOA11a]
GO:0016798 - hydrolase activity, acting on glycosyl bonds Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005576 - extracellular region Inferred from experiment Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, DiazMejia09, Park99]

MultiFun Terms: metabolism degradation of macromolecules polysaccharides

Essentiality data for bcsZ knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]

Credits:
Last-Curated ? 25-Jun-2014 by Keseler I , SRI International


Enzymatic reaction of: endo-1,4-D-glucanase

Synonyms: CMCase, β-1,4-D-glucan glucanohydrolase

EC Number: 3.2.1.4

cellulose[extracellular space] + n H2O[extracellular space] <=> n a cellodextrin[extracellular space]

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is physiologically favored in the direction shown.

Summary:
Enzyme activity was assayed using carboxymethyl cellulose (CMC) as a substrate [Park99].

T(opt): 40 °C [Park99]

pH(opt): 7 [Park99]


Sequence Features

Feature Class Location Citations Comment
Signal-Sequence 1 -> 21
[UniProt10a]
UniProt: Non-Experimental Qualifier: potential;
Chain 22 -> 368
[UniProt09]
UniProt: Endoglucanase;
Active-Site 55
[UniProt10a]
UniProt: Proton donor; Non-Experimental Qualifier: by similarity;
Active-Site 116
[UniProt10a]
UniProt: Nucleophile; Non-Experimental Qualifier: potential;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
Gene common name added 21-July-99. BcsC gene identified as an endo-1,4-D-glucanase, EC# 3.2.1.4. Ref: [99200480] Mol Gen Genet 1999 261:236-41. Park YW and Yun HD.
10/20/97 Gene b3531 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG12258; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Blattner97: Blattner FR, Plunkett G, Bloch CA, Perna NT, Burland V, Riley M, Collado-Vides J, Glasner JD, Rode CK, Mayhew GF, Gregor J, Davis NW, Kirkpatrick HA, Goeden MA, Rose DJ, Mau B, Shao Y (1997). "The complete genome sequence of Escherichia coli K-12." Science 277(5331);1453-74. PMID: 9278503

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA01a: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

Gonnet04: Gonnet P, Rudd KE, Lisacek F (2004). "Fine-tuning the prediction of sequences cleaved by signal peptidase II: a curated set of proven and predicted lipoproteins of Escherichia coli K-12." Proteomics 4(6);1597-613. PMID: 15174130

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Mazur11: Mazur O, Zimmer J (2011). "Apo- and cellopentaose-bound structures of the bacterial cellulose synthase subunit BcsZ." J Biol Chem 286(20);17601-6. PMID: 21454578

Omadjela13: Omadjela O, Narahari A, Strumillo J, Melida H, Mazur O, Bulone V, Zimmer J (2013). "BcsA and BcsB form the catalytically active core of bacterial cellulose synthase sufficient for in vitro cellulose synthesis." Proc Natl Acad Sci U S A 110(44);17856-61. PMID: 24127606

Park99: Park YW, Yun HD (1999). "Cloning of the Escherichia coli endo-1,4-D-glucanase gene and identification of its product." Mol Gen Genet 261(2);236-41. PMID: 10102357

Salmon03: Salmon K, Hung SP, Mekjian K, Baldi P, Hatfield GW, Gunsalus RP (2003). "Global gene expression profiling in Escherichia coli K12. The effects of oxygen availability and FNR." J Biol Chem 278(32);29837-55. PMID: 12754220

Serra13: Serra DO, Richter AM, Hengge R (2013). "Cellulose as an architectural element in spatially structured Escherichia coli biofilms." J Bacteriol 195(24);5540-54. PMID: 24097954

Sofia94: Sofia HJ, Burland V, Daniels DL, Plunkett G, Blattner FR (1994). "Analysis of the Escherichia coli genome. V. DNA sequence of the region from 76.0 to 81.5 minutes." Nucleic Acids Res 1994;22(13);2576-86. PMID: 8041620

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

Zogaj01: Zogaj X, Nimtz M, Rohde M, Bokranz W, Romling U (2001). "The multicellular morphotypes of Salmonella typhimurium and Escherichia coli produce cellulose as the second component of the extracellular matrix." Mol Microbiol 39(6);1452-63. PMID: 11260463

Other References Related to Gene Regulation

Huerta03: Huerta AM, Collado-Vides J (2003). "Sigma70 promoters in Escherichia coli: specific transcription in dense regions of overlapping promoter-like signals." J Mol Biol 333(2);261-78. PMID: 14529615


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Wed Dec 17, 2014, biocyc13.