Cellulose production by Enterobacter sp. CJF-002 and identification of genes for cellulose biosynthesis

Enterobacter sp. CJF-002, which had been isolated as a cellulose producer with saccharides as a carbon source, was shown to efficiently produce cellulose from beet molasses (B-Mol) and biodiesel fuel by-product (BDF-B), renewable non-edible and inexpensive biomasses. The cellulose production rates of Enterobacter sp. CJF-002 using B-Mol and BDF-B as carbon sources were faster than those of Acetobacter xylinum (A. xylinum) ATCC23769, a representative cellulose producing bacterium. To clarify the biosynthetic machinery of cellulose in the strain, genes responsible for cellulose biosynthesis were cloned. Six open reading frames (ORFs) were suggested to be clustered and their amino acid sequences had high similarities with those of BcsA, BcsB, BcsZ (endoglucanase), BcsC, YhjQ, and YhjK from Escherichia coli, respectively. Of these, the former four genes showed low similarities to corresponding orthologs in a cellulose biosynthetic gene cluster of A. xylinum. A bcsC-knockout mutant produced no cellulose, confirming that the gene is essential for cellulose production of Enterobacter sp. CJF-002. The predicted three-dimensional structure of BcsZ_En from Enterobacter sp. CJF-002 had high similarity with that of CMCax (endoglucanase) from A. xylinum ATCC23769 in spite of the low similarity in their amino acid sequences. Taken together, A. xylinum and Enterobacter sp. CJF-002 might produce cellulose via a similar synthetic mechanism.