A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia's hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medi...A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia's hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medium (MSM). The extreme thermophile Brevibacillus sp. PLI-1 was found to produce extracellular thermophilic alkaline protease with optimal activity at 70℃ and pH 8.0-9.0. The molecular weight of the protease was estimated to be around 56 kD by SDS-PAGE. The maximum activity of the protease was 26.54 U mL-1. The protease activity did not decrease after 30 min and still retained more than 70% of relative activity after 60 min at 70℃ and pH 8.0. The ion Mg2+ was found to promote protease activity at both low and high concentrations, whereas Cu2+ and Zn2+ could almost completely inhibit the activity. Divalent cation chelator EDTA inhibited the enzyme activity by 55.06% ± 0.27%, while the inhibition caused by PMSF, Leupeptin, Pepstain A and Benzamidine were 66.78% ± 3.25%, 52.37% ± 0.25%, 62.47% ± 2.96% and 50.99% ± 0.24%, respectively. Based on these observations, the enzyme activity was conspicuously sensitive to the serine and cysteine protease inhibitors. All these results indicated that the protease isolated from the strain PLI-1 was a thermophilic protease and had a high-temperature stability and a pH stability.展开更多
Anaerobic oxidation of methane and sulfate reduction was studied in the pore waters of four cores at two stations of the middle Okinawa Trough. Pore water vertical distributions of sulfate, methane, sulfide, total alk...Anaerobic oxidation of methane and sulfate reduction was studied in the pore waters of four cores at two stations of the middle Okinawa Trough. Pore water vertical distributions of sulfate, methane, sulfide, total alkalinity, ammonium, and phosphate were determined in this study. Our results show strong linear sulfate concentration gradients of 6.83 mmol/L m?1 in Core A and 5.96 mmol/L m?1 in Core C, which were collected from two stations. Concurrent variations of methane, total alkalinity and hydrogen sulfide all exhibit steep increases with depth at both cores, which indicate active methane seep activities around two stations. Pore water ammonium and phosphate concentrations reveal minor influences of organic matter degradation on sulfate reduction at two stations. Sulfate methane interface(SMI) was extrapolated from linear sulfate profiles in methane seep cores. Shallower SMI depths(A: 4.9 mbsf; C: 5.4 mbsf) indicate strong methane fluxes and active anaerobic oxidation of methane in the underlying sediments.展开更多
基金supported by the Basic Scientific Research Foundation of FIO, SOA (2010G13)the National Natural Science Foundation of China (41076108)+1 种基金the Science Foundation for Youth Scholars of State Oceanic Administration (2012131)the Public Welfare Project of Marine Science Research (200805039)
文摘A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia's hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medium (MSM). The extreme thermophile Brevibacillus sp. PLI-1 was found to produce extracellular thermophilic alkaline protease with optimal activity at 70℃ and pH 8.0-9.0. The molecular weight of the protease was estimated to be around 56 kD by SDS-PAGE. The maximum activity of the protease was 26.54 U mL-1. The protease activity did not decrease after 30 min and still retained more than 70% of relative activity after 60 min at 70℃ and pH 8.0. The ion Mg2+ was found to promote protease activity at both low and high concentrations, whereas Cu2+ and Zn2+ could almost completely inhibit the activity. Divalent cation chelator EDTA inhibited the enzyme activity by 55.06% ± 0.27%, while the inhibition caused by PMSF, Leupeptin, Pepstain A and Benzamidine were 66.78% ± 3.25%, 52.37% ± 0.25%, 62.47% ± 2.96% and 50.99% ± 0.24%, respectively. Based on these observations, the enzyme activity was conspicuously sensitive to the serine and cysteine protease inhibitors. All these results indicated that the protease isolated from the strain PLI-1 was a thermophilic protease and had a high-temperature stability and a pH stability.
基金supported by the National Natural Science Foundation of China(Grand Nos.41306062,41104086)the Key Laboratory of Gas Hydrate Foundation(Grand Nos.SHW[2014]-DX-03,SHW[2014]-DX-04)the Key Laboratory of Marine Hydrocarbon Resources and Environmental Geology Foundation(Grand No.MRE 201213)
文摘Anaerobic oxidation of methane and sulfate reduction was studied in the pore waters of four cores at two stations of the middle Okinawa Trough. Pore water vertical distributions of sulfate, methane, sulfide, total alkalinity, ammonium, and phosphate were determined in this study. Our results show strong linear sulfate concentration gradients of 6.83 mmol/L m?1 in Core A and 5.96 mmol/L m?1 in Core C, which were collected from two stations. Concurrent variations of methane, total alkalinity and hydrogen sulfide all exhibit steep increases with depth at both cores, which indicate active methane seep activities around two stations. Pore water ammonium and phosphate concentrations reveal minor influences of organic matter degradation on sulfate reduction at two stations. Sulfate methane interface(SMI) was extrapolated from linear sulfate profiles in methane seep cores. Shallower SMI depths(A: 4.9 mbsf; C: 5.4 mbsf) indicate strong methane fluxes and active anaerobic oxidation of methane in the underlying sediments.
