Paenibacillus sp. strain HC1 xylanases responsible for degradation of rice bran hemicellulose.

Paenibacillus sp. strain HC1 is the first bacterium capable of growing on rice bran hemicellulose as a sole carbon source. Two xylanases (Xyl-I and -II) were purified from the bacterial culture fluid and enzymatically characterized. Xyl-I and -II showed monomer forms with molecular masses of 30 and 18kDa, respectively, and were most active at around pH 5.0 and 45 degrees C. Xylooligosaccharides were degraded to xylobiose and xylose by Xyl-I, but not by Xyl-II, suggesting that Xyl-I plays an important role in complete depolymerization of xylan. Both enzymes acted endolytically on rice bran hemicellulose, indicating that Xyl-I and -II contribute to the structure determination and practical use of the polysaccharide, an unutilized biomass in technology.

Sequence and analysis of the 46.6-kb plasmid pA1 from Sphingomonas sp. A1 that corresponds to the typical IncP-1beta plasmid backbone without any accessory gene.

Sphingomonas sp. A1 (strain A1) is capable of directly incorporating macromolecules (e.g., alginate) through the specialized import system--"super-channel." Here, we report the complete DNA sequence and genetic organization of plasmid pA1 from strain A1. Nucleotide sequence analysis revealed that pA1 comprises 46,557 bp encoding 49 open reading frames (ORFs) with 65% G+C content and abundant GCCG/CGGC motifs. Many predicted pA1 ORFs showed high similarity to pA81 ORFs; pA81 is supposedly a self-transmissible promiscuous incompatibility (Inc) group P-1beta plasmid. Unlike any reported IncP-1 plasmids, pA1 contains no inserted mobile genetic elements. The genetic organization and predicted pA1 ORFs showed greater similarity to the IncP-1beta plasmid backbone than to the IncP-1alpha plasmid backbone. pA1 contains restriction site-associated repeat sequences typical of the IncP-1beta but absent in the IncP-1alpha and delta subgroups. Thus, the overall pA1 structure corresponds to that of the typical IncP-1beta plasmids. Phylogenetic analysis of the replication-associated proteins suggested that pA1 may have diverged later along with the two IncP-1beta plasmids--pA81 and pB4. The 2.4-kb duplicates of stable inheritance genes klcAB and korC in pA1 possibly resulted from insertion and/or recombination events via the repeat sequences flanking these duplicates.

Engineered membrane superchannel improves bioremediation potential of dioxin-degrading bacteria.

Sphingomonas sp. A1 possesses specialized membrane structures termed 'superchannels' that enable the direct incorporation of macromolecules into the cell. We have engineered two related sphingomonads, the dioxin-degrading S. wittichii RW1 and the polypropylene glycol-degrading S. subarctica IFO 16058(T), to incorporate this superchannel into their cell membranes. In both cases the bioremediation capability of the organisms was substantially increased pointing at the potential of this approach as a general strategy to improve bacterial degradation of hazardous compounds in the environment.

Degradation of rice bran hemicellulose by Paenibacillus sp. strain HC1: gene cloning, characterization and function of beta-D-glucosidase as an enzyme involved in degradation.

A bacterium (strain HC1) capable of assimilating rice bran hemicellulose was isolated from a soil and identified as belonging to the genus Paenibacillus through taxonomical and 16S rDNA sequence analysis. Strain HC1 cells grown on rice bran hemicellulose as a sole carbon source inducibly produced extracellular xylanase and intracellular glycosidases such as beta-D-glucosidase and beta-D-arabinosidase. One of them, beta-D-glucosidase, was further analyzed. A genomic DNA library of the bacterium was constructed in Escherichia coli and gene coding for beta-D-glucosidase was cloned by screening for beta-D-glucoside-degrading phenotype in E. coli cells. Nucleotide sequence determination indicated that the gene for the enzyme contained an open reading frame consisting of 1,347 bp coding for a polypeptide with a molecular mass of 51.4 kDa. The polypeptide exhibits significant homology with other bacterial beta-D-glucosidases and belongs to glycoside hydrolase family 1. Beta-D-Glucosidase purified from E. coli cells was a monomeric enzyme with a molecular mass of 50 kDa most active at around pH 7.0 and 37 degrees C. Strain HC1 glycosidases responsible for degradation of rice bran hemicellulose are expected to be useful for structurally determining and molecularly modifying rice bran hemicellulose and its derivatives.