A novel thermophilic endoglucanase (EGt) was extracted from a mesophilic fungus (Fusarium oxysporum L19). We invoked conventional kinetic enzyme reactions using the sodium salt of carboxymethyl cellulose (CMC-Na) as s...A novel thermophilic endoglucanase (EGt) was extracted from a mesophilic fungus (Fusarium oxysporum L19). We invoked conventional kinetic enzyme reactions using the sodium salt of carboxymethyl cellulose (CMC-Na) as substrate. EGt displayed optimal activity at 75℃ when kept running 30 min in the temperature range of 30―85℃. Thermal stability curve measured at 70℃ suggested that its half-life time is 15.1 min. The activity was enhanced in the presence of Co2+ or Mg2+ but inhibited by Pb2+ and Fe3+. Moreover, N-bromosuccinimide (NBS) modification resulted in a complete loss of EGt activity, suggesting that tryptophan residues may be involved in the enzyme active site. Amino acid composition analysis demonstrated that EGt contains more proline residues. EGt lacks activity towards p-nitro- phenyl cellobiose (pNPC). The N-terminal amino acid se- quence of EGt is SYRVPAANGFPNPDASQEKQ, and the gene of EGt was sequenced and analyzed. Extensive se- quence alignments failed to show any homology between EGt and any known endoglucanases. This is the first report addressing the thermal adaptation of a cellulolytic enzyme from the mesophilic fungus F. oxysporum. Maybe the ex- pression of multiple isoenzyme in an organism helps it adapt to complex living environments.展开更多
The application of the response surface methodology and the central composite design（CCD） technique for modeling and optimization of the influence of some operating variables on copper,molybdenum and rhenium recover...The application of the response surface methodology and the central composite design（CCD） technique for modeling and optimization of the influence of some operating variables on copper,molybdenum and rhenium recoveries in a bioleaching process was investigated.Three main bioleaching parameters,namely pH,solid concentration and inoculum percent,were changed during the bioleaching tests based on CCD.The ranges of the bioleaching process variables used in the design were as follows：pH1.46-2.14,solid concentration 0.95%-11.05%,and inoculum percent 1.59%-18.41%.A total of 20 bioleaching tests were carried out by the CCD method according to software-based designed matrix.Empirical model equations were developed according to the copper,molybdenum and rhenium recoveries obtained with these three parameters.Model equations of responses at the base of parameters were achieved by using statistical software.The model equations were then individually optimized by using quadratic programming to maximize copper,molybdenum and rhenium recoveries individually within the experimental range.The optimum conditions for copper recovery were pH 1.68,solid concentration 0.95% and the inoculum 18.41%（v/v）,while molybdenum and rhenium recoveries were 2.18% and 24.41%,respectively.The predicted values for copper,molybdenum and rhenium recoveries were found to be in good agreement with the experimental values.Also jarosite formation during bioleaching tests was also investigated.展开更多
基金We acknowledge the help of Robert Koebner in the linguistic correction of the manuscript.This work was supported by the National Natural Science Foundation of China(Grants Nos.30370013&30470051)the Major State Basic Research Development Research Program of China("973"Project)(Grant Nos.2003CB716006&2004CB719702).
文摘A novel thermophilic endoglucanase (EGt) was extracted from a mesophilic fungus (Fusarium oxysporum L19). We invoked conventional kinetic enzyme reactions using the sodium salt of carboxymethyl cellulose (CMC-Na) as substrate. EGt displayed optimal activity at 75℃ when kept running 30 min in the temperature range of 30―85℃. Thermal stability curve measured at 70℃ suggested that its half-life time is 15.1 min. The activity was enhanced in the presence of Co2+ or Mg2+ but inhibited by Pb2+ and Fe3+. Moreover, N-bromosuccinimide (NBS) modification resulted in a complete loss of EGt activity, suggesting that tryptophan residues may be involved in the enzyme active site. Amino acid composition analysis demonstrated that EGt contains more proline residues. EGt lacks activity towards p-nitro- phenyl cellobiose (pNPC). The N-terminal amino acid se- quence of EGt is SYRVPAANGFPNPDASQEKQ, and the gene of EGt was sequenced and analyzed. Extensive se- quence alignments failed to show any homology between EGt and any known endoglucanases. This is the first report addressing the thermal adaptation of a cellulolytic enzyme from the mesophilic fungus F. oxysporum. Maybe the ex- pression of multiple isoenzyme in an organism helps it adapt to complex living environments.
基金supported by the National Iranian Copper Industry Co. and Geological Survey of Iran
文摘The application of the response surface methodology and the central composite design（CCD） technique for modeling and optimization of the influence of some operating variables on copper,molybdenum and rhenium recoveries in a bioleaching process was investigated.Three main bioleaching parameters,namely pH,solid concentration and inoculum percent,were changed during the bioleaching tests based on CCD.The ranges of the bioleaching process variables used in the design were as follows：pH1.46-2.14,solid concentration 0.95%-11.05%,and inoculum percent 1.59%-18.41%.A total of 20 bioleaching tests were carried out by the CCD method according to software-based designed matrix.Empirical model equations were developed according to the copper,molybdenum and rhenium recoveries obtained with these three parameters.Model equations of responses at the base of parameters were achieved by using statistical software.The model equations were then individually optimized by using quadratic programming to maximize copper,molybdenum and rhenium recoveries individually within the experimental range.The optimum conditions for copper recovery were pH 1.68,solid concentration 0.95% and the inoculum 18.41%（v/v）,while molybdenum and rhenium recoveries were 2.18% and 24.41%,respectively.The predicted values for copper,molybdenum and rhenium recoveries were found to be in good agreement with the experimental values.Also jarosite formation during bioleaching tests was also investigated.
