Engineering of RHA1 for utilisation of carboxymethylcellulose.

RHA1 was engineered to utilise the cellulose component of lignocellulose, as well as the lignin fraction, by introduction of cellulase genes. The genome of RHA1 was found to contain two β-glucosidase genes, RHA1_ro01034 and RHA1_ro02947, which support growth on cellobiose as growth substrate. Five Gram-positive endocellulase genes and one exocellulase gene were cloned into expression vector pTipQC2, and expressed in RHA1. Endoglucanase activity was detected, with highest activity using , and this recombinant strain grew on minimal media containing 0.5% carboxymethylcellulose (CMC). The RHA1 genome was also found to contain a 3-dehydroshikimate dehydratase gene RHA1_ro01367, which supports growth on quinic acid as growth substrate, and conversion to protocatechuic acid. Therefore, this bacterium shows promise for further engineering to utilise cellulose for conversion to protocatechuic acid-derived bioproducts.