PUL ID

PUL0311

PubMed

31455320, BMC Biotechnol. 2019 Aug 27;19(1):63. doi: 10.1186/s12896-019-0556-0.

Characterization method

enzyme activity assay

Genomic accession number

HG738867.1

Nucelotide position range

3086037-3095373

Substrate

cellulose

Loci

bcsABZC

Species

Escherichia coli/562

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

bcsA BN896_3222 CDJ73343.1 0 - 2619 (+) HG738867.1:3086037-3088656 -
bcsB BN896_3221 CDJ73344.1 2629 - 4969 (+) HG738867.1:3088666-3091006 -
bcsZ BN896_3220 CDJ73345.1 4975 - 6082 (+) HG738867.1:3091012-3092119 -
bcsC BN896_3219 CDJ73346.1 6114 - 9337 (+) HG738867.1:3092151-3095374 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

bcsA 1 - 2619 (+) CAZyme: GT2|GT2_Glycos_transf_2 Yes
bcsB 2630 - 4969 (+) TC: gnl|TC-DB|P37652|4.D.3.1.6 Yes
bcsZ 4976 - 6082 (+) CAZyme: GH8 Yes
bcsC 6115 - 9337 (+) TC: gnl|TC-DB|P37650|1.B.55.3.1 Yes

PUL ID

PUL0311

PubMed

31455320, BMC Biotechnol. 2019 Aug 27;19(1):63. doi: 10.1186/s12896-019-0556-0.

Title

Identification and characterization of an Endo-glucanase secreted from cellulolytic Escherichia coli ZH-4.

Author

Pang J, Wang J, Liu Z, Zhang Q, Qi Q

Abstract

BACKGROUND: In the previous study, the cellulolytic Escherichia coli ZH-4 isolated from bovine rumen was found to show extracellular cellulase activity and could degrade cellulose in the culture. The goal of this work was to identify and characterize the secreted cellulase of E. coli ZH-4. It will be helpful to re-understand E. coli and extend its application in industry. RESULTS: A secreted cellulase was confirmed to be endo-glucanase BcsZ which was encoded by bcsZ gene and located in the cellulose synthase operon bcsABZC in cellulolytic E. coli ZH-4 by western blotting. Characterization of BcsZ indicated that a broad range of pH and temperature tolerance with optima at pH 6.0 and 50 degrees C, respectively. The apparent Michaelis-Menten constant (K(m)) and maximal reaction rate (V(max)) for BcsZ were 8.86 mg/mL and 0.3 muM/min.mg, respectively. Enzyme activity of BcsZ was enhanced by Mg(2+) and inhibited by Zn(2+), Cu(2+) and Fe(3+). BcsZ could hydrolyze carboxymethylcellulose (CMC) to produce cello-oligosaccharides, cellotriose, cellobiose and glucose. CONCLUSIONS: It is confirmed that extracellular cellulolytic capability of E. coli ZH-4 was attributed to BcsZ, which explained why E. coli ZH-4 can grow on cellulose. The endo-glucanase BcsZ from E. coli-ZH4 has some new characteristics which will extend the understanding of endo-glucanase. Analysis of the secretion characteristics of BcsZ provided a great reference for applying E. coli in multiple industrial fields.