Yoshiki Matsumiya

Phylogenetic analysis of long-chain hydrocarbon-degrading bacteria and evaluation of their hydrocarbon-degradation by the 2,6-DCPIP assay

Thirty-six bacteria that degraded long-chain hydrocarbons were isolated from natural environments using long-chain hydrocarbons (waste car engine oil, base oil or the c-alkane fraction of base oil) as the sole carbon and energy source. A phylogenetic tree of the isolates constructed using their 16S rDNA sequences revealed that the isolates were divided into six genera plus one family (Acinetobacter, Rhodococcus, Gordonia, Pseudomonas, Ralstonia, Bacillus and Alcaligenaceae, respectively). Furthermore, most of the isolates (27 of 36) were classified into the genera Acinetobacter, Rhodococcus or Gordonia. The hydrocarbon-degradation similarity in each strain was confirmed by the 2,6-dichlorophenol indophenol (2,6-DCPIP) assay. Isolates belonging to the genus Acinetobacter degraded long-chain normal alkanes (n-alkanes) but did not degrade short-chain n-alkanes or cyclic alkanes (c-alkanes), while isolates belonging to the genera Rhodococcus and Gordonia degraded both long-chain n-alkanes and c-alkanes.

Isolation and characterization of an ether‐type polyurethane‐degrading micro‐organism and analysis of degradation mechanism by Alternaria sp.

Aims:  To degrade ether‐type polyurethane (ether‐PUR), ether‐PUR–degrading micro‐organism was isolated. Moreover, ether‐PUR–degrading mechanisms were analysed using model compounds of ether‐PUR.

Mutational effect for stability in a conserved region of thermolysin

Aims:  To investigate the mutational effect for the stability of thermolysin (TLN) in conserved regions.

Soybean Peptide: Novel Plant Growth Promoting Peptide from Soybean

Soybean is one of the most important agricultural products and its global production was more than 200 million tons per year in 2005 (Table 1) (Ministry of Agriculture, Forestry and Fisheries (Japan), 2007; Uchida, 2007). Soybean is used mainly as a vegetable oil (31.6 million tons a year in 2005) and the production ratio is the highest (30%) among vegetable oils. Soybean waste, which remains after extraction of vegetable oil, contains about 50% proteins, which consist of a well-balanced mix of amino acids. Therefore, soybean waste is a valuable biomass for animal feedstuffs. Soybean is used directly as food in Japan and several Asian countries but soybean proteins are used less widely elsewhere in the world.

Expression in Escherichia coli, Refolding, and Purification of the Recombinant Mature Form of Human Matrix Metalloproteinase 7 (MMP-7)

In the latent pro-form of matrix metalloproteinase 7 (MMP-7), the cysteine residue in the pro-peptide binds the active-site zinc ion. Hence, recombinant active MMP-7 was prepared from pro-MMP-7 by modification of this cysteine residue with a mercuric reagent. In this study, mature MMP-7 was expressed in Escherichia coli as inclusion bodies, solubilized, and refolded with 1 M L-arginine. The purified product was indistinguishable from the one prepared from pro-MMP-7 as assessed by hydrolysis of (7-methoxycoumarin-4-yl)acetyl-L-Pro-L-Leu-Gly-L-Leu-[N 3-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl]-L-Ala-L-Arg-NH2.

Isolation and characterization of a bluegill-degrading microorganism, and analysis of the root hair-promoting effect of the degraded products.

Bluegill-degrading bacteria were isolated from various environmental sources. Brevibacillus sp. BGM1 degraded bluegill efficiently at 50 °C, and its culture supernatant showed the highest peptide and amino acid concentrations as trichloroacetic acid (TCA) soluble fraction (ASF) (10.7 mg/ml) of all supernatants obtained with bluegill as a substrate. Strain BGM1 secreted a protease(s) into the medium, and the concentration of peptides and amino acids gradually increased. The fertile effect of the degraded bluegill products (DGP) on Brassica rapa was also investigated. The root hair density of B. rapa grown with DGP at a concentration of 30 μg peptides and amino acids/ml was about 1.7 times higher than when grown with the same concentration of undegraded bluegill. DGP was shown to increase root hair numbers and adventitious root formation. The results of this study suggest that a specific peptide(s) for promotion of root hair is produced from the order Perciformes with a protease(s) from BGM1.

Analysis of peptide uptake and location of root hair‐promoting peptide accumulation in plant roots

Peptide uptake by plant roots from degraded soybean‐meal products was analyzed in Brassica rapa and Solanum lycopersicum. B. rapa absorbed about 40% of the initial water volume, whereas peptide concentration was decreased by 75% after 24 h. Analysis by reversed‐phase HPLC showed that number of peptides was absorbed by the roots during soaking in degraded soybean‐meal products for 24 h. Carboxyfluorescein‐labeled root hair‐promoting peptide was synthesized, and its localization, movement, and accumulation in roots were investigated. The peptide appeared to be absorbed by root hairs and then moved to trichoblasts. Furthermore, the peptide was moved from trichoblasts to atrichoblasts after 24 h. The peptide was accumulated in epidermal cells, suggesting that the peptide may have a function in both trichoblasts and atrichoblasts. Copyright