To generate transgenic plants capable of utilizing exogenous phytate, an Aspersgillus fumigatus phytase gene （fphyA） was constitutively expressed in tobacco and recombinant enzyme was secreted from plant roots into ...To generate transgenic plants capable of utilizing exogenous phytate, an Aspersgillus fumigatus phytase gene （fphyA） was constitutively expressed in tobacco and recombinant enzyme was secreted from plant roots into the rhizosphere using the signal sequence from tobacco calreticulin. After 40 days of plant growth in hydroponic media, phytase activities in leaves, stems, roots and growth media of transgenic plants were 8.6-fold, 7.4-fold, 12.6-fold and 14.3-fold higher than those of wild-type plants. Signifi- cant improvements in plant growth and phosphorus （P） utilization were observed in the transgenic plants. When phytate was supplied as the sole P source, 45-day-old transgenic tobaccos accumulated 1.0-fold and 0.5-fold more shoot and root biomass than wild-type tobaccos, with a concomitant of l. 7-fold increase in total P concentration. These results indicate that secretive expression of the A. fumigatus phytase improves acquisition and use of P from phytate in plants.展开更多
A new butyrolactone derivative,namely butyrolactone Ⅷ （1）,and six known butyrolactones （2-7） were separated from the ethyl acetate （EtOAc） extract of the fermentation broth of a fungus,Aspergillus terreus MXH-2...A new butyrolactone derivative,namely butyrolactone Ⅷ （1）,and six known butyrolactones （2-7） were separated from the ethyl acetate （EtOAc） extract of the fermentation broth of a fungus,Aspergillus terreus MXH-23.The chemical structures of these metabolites were identified by analyzing their nuclear magnetic resonance （NMR） and mass spectrometry （MS）.Known butyrolactone derivatives contain an α,β-unsaturated γ-lactone ring with α-hydroxyl and y-benzyl,and butyrolactone Ⅷ （1） was the first butyrolactones contains α-benzyl and γ-hydroxyl on α,β-unsaturated lactone ring.All of the butyrolactone derivatives were tested for their anti-influenza （H 1N 1） effects.Derivatives 4 and 7 showed moderate antiviral activities while the newly-identified,derivative 1,did not.展开更多
Aspergillus oryzae 3042 mycelium pellets were immobilized by crosslinking method with reagents of gelatin and formaldehyde. An orthogonal design table was used to determine the optimal immobilization conditions. The L...Aspergillus oryzae 3042 mycelium pellets were immobilized by crosslinking method with reagents of gelatin and formaldehyde. An orthogonal design table was used to determine the optimal immobilization conditions. The L-aminoacylase activity of immobilized mycelium pellets under optimal conditions was assayed. The results show that the L-aminoacylase activity was 1350 U·g^-1 and the activity retention rate was 83%. The properties of the immobilized preparation were studied. Compared with free pellets, the appropriate pH of reaction system changed from 7.0 to 8.0, temperature changed from 52℃ to 63℃, and the ranges of the optimum reaction conditions were all improved. The effect of Co^2＋ on immobilized mycelium pellets was investigated and the favorable concentration was determined. When the immobilized preparation was used for the resolution of D, L-alanine in a packed bed reactor with 10 mm in diameter, 200 mm in height, the operational stability was increased, and the half-life period was 53 d. Thermal stability analysis shows that the immobilized pellets were more stable than the free pellets.展开更多
A strain HB-03 to produce alkaline extracellular lipase was isolated from oil-rich soil samples and identified as Aspergillus awamori. The growth conditions and nutritional factors for lipase production by strain HB-0...A strain HB-03 to produce alkaline extracellular lipase was isolated from oil-rich soil samples and identified as Aspergillus awamori. The growth conditions and nutritional factors for lipase production by strain HB-03 were optimized, and the maximum lipase production of （45.9±2.3） U/mL was obtained at 30 ℃ and pH 7.0 after 36 h using olive oil （1%） and sucrose （0.5%） as carbon sources and combination of peptone （2%）, yeast extract （0.5%） and ammonium sulfate （0.1%） as nitrogen sources. The lipase was purified to homogeneity with 10.6-fold, 18.84% yield and a specific activity of 1 862.2 U/mg using ammonium sulfate precipitation followed by SephadexG-75 gel filtration chromatography. The purified lipase with molecular mass of 68 ku was estimated by SDS-PAGE. The optimum pH and temperature for the purified lipase were found to be 8.5 and 40 ℃, respectively. The lipase kept more than 80% of activity in pH 7.0-10.0 and temperatures up to 45 ℃. The metal ions of Mn2＋, Ba2＋ significantly enhanced the lipase activity, whereas Cu2＋, Fe3＋ and Mg2＋ strongly reduced the lipase activity. The Km and Vmax values of the purified enzyme for p-nitrophenyl palmitate were 0.13 mrnol/L and 60.6 mmol/（L.min）, respectively. The results show that this novel lipase has potential industrial applications.展开更多
Phosphorus （P） is a major growth-limiting nutrient, and unlike the case of nitrogen （N）, there is no large atmospheric source that can be made biologically available. Moreover, P governs crucial role in plant as i...Phosphorus （P） is a major growth-limiting nutrient, and unlike the case of nitrogen （N）, there is no large atmospheric source that can be made biologically available. Moreover, P governs crucial role in plant as it stimulates root development and growth, gives plant rapid and vigorous start leading to better tillering and essential for many metabolic processes for seed formation. Soil microbes play very important role in bio-weathering and biodegradation. The microorganisms produce low molecular mass organic acids, which attack the phosphate structure and transform phosphorus from non-utilizable to the utilizable for the plants form. The test of the relative efficiency of isolated strains is carried out by selecting the microorganisms that are capable of producing a halo/clear zone on a plate owing to the production of organic acids into the surrounding medium. It is a well-known fact that as the particle size of rock phosphate decreases, the microbe mediated solubilization of rock phosphate increases in soil. In the present investigation, microbial solubilization of nano rock phosphate （〈 100 nm） particles was studied. Experimental results revealed that Pseudomonas striata solubilized 11.45% of the total P after 24 h of incubation from nano rock phosphate particles while 28.95% and 21.19% of the total P was solubilized by Aspergillus niger （black pigmented） and Aspergillus niger （green pigmented）, respectively. It was also observed that Aspergillus niger has the higher ability to dissolve Udaipur rock phosphate than Pseudomonas striata.展开更多
The objective of this study is to improve the production of L-DOPA from fungal source like Aspergillus terreus that can be further used to large-scale commercial production of this important drug from microbial source...The objective of this study is to improve the production of L-DOPA from fungal source like Aspergillus terreus that can be further used to large-scale commercial production of this important drug from microbial sources. L-DOPA, a dopamine precursor that can pass the blood-brain barrier, is presently the drug of choice for Parkinson＇s disease. Microbial production and isolation of L-DOPA from natural sources is yet to be achieved an economical process. In this study, the mycelial pellets ofAspergillus terreus 104 were entrapped in 2% calcium alginate and were studied for their capacity for L-3, 4-dihydroxyphenylalanine production. Results showed that the immobilized pellets produced L-DOPA to the extent of 0.74 mg·G^-1 biomass while the free pellets produced 0.8 mg·G^-1 biomass. Further, storage of immobilized pellets for 96 h at 4 ℃ resulted in the reduction of the original L-DOPA producing activity of the gel beads only 40% and that of free pellets lost completely. In order to improve the production yield, further experiments were designed. It was found that L-DOPA production could be prolonged with repeated batch wise use of immobilized mycelial pellets in calcium alginate retaining 80% of their L-DOPA producing capacity for a period of 72 h while free pellets lost completely within 24 h. Results of this kind therefore is interesting and promising for commercial scale production of L-DOPA from microbial sources.展开更多
We investigated the differences in the gene expression of carbonic anhydrase （CA） and cysteine synthase （CysM） between two weathering conditions, with either soluble potassium or insoluble potassium. We cultured a...We investigated the differences in the gene expression of carbonic anhydrase （CA） and cysteine synthase （CysM） between two weathering conditions, with either soluble potassium or insoluble potassium. We cultured a strain of A. niger by adopting a variant Czapek medium （using NazHPO4 as a substitute for KzHPO4） in two groups, Group A （containing silicate minerals bearing potassium but without KC1） and Group B （with KCI） . We extracted the mRNAs of CA and CysM from these two groups and performed real-time quantitative polymerase chain reactions （RT-qPCR）. We constructed relative standard curves by using glyceraldehyde-3-phosphate dehydrogenase （GAPDH） as the reference to confirm a consistent amplification effi- ciency of the target genes （CA and CysM） and the reference gene and quantified the gene expression of the targets in a relative manner. Our results showed that CA and CysM in Group A were upregulated for 1.7 times and 11.7 times, respectively, com- pared with those in Group B. Furthermore, we also analyzed some metabolic pathways and functions of the A. niger-induced weathering of potassium-bearing minerals, which involved the synthesizing of these two enzymes. Thus our work provides materials for further study of the roles of A. niger in the metabolic regulation during the weathering process of potassi- um-beating minerals.展开更多
文摘To generate transgenic plants capable of utilizing exogenous phytate, an Aspersgillus fumigatus phytase gene （fphyA） was constitutively expressed in tobacco and recombinant enzyme was secreted from plant roots into the rhizosphere using the signal sequence from tobacco calreticulin. After 40 days of plant growth in hydroponic media, phytase activities in leaves, stems, roots and growth media of transgenic plants were 8.6-fold, 7.4-fold, 12.6-fold and 14.3-fold higher than those of wild-type plants. Signifi- cant improvements in plant growth and phosphorus （P） utilization were observed in the transgenic plants. When phytate was supplied as the sole P source, 45-day-old transgenic tobaccos accumulated 1.0-fold and 0.5-fold more shoot and root biomass than wild-type tobaccos, with a concomitant of l. 7-fold increase in total P concentration. These results indicate that secretive expression of the A. fumigatus phytase improves acquisition and use of P from phytate in plants.
基金supported by National Natural Science Foundation of China (Nos.41176120 and 30973627)the National High Technology Research and Development Program of China (No.2013AA092901)+3 种基金the Program for New Century Excellent Talents in University (No.NCET12-0499)Promotive Research Fund for Excellent Young and Middle-Aged Scientists of Shandong Province (BS 2010HZ027)the Public Projects of State Oceanic Administration (No. 2010418022-3)the Program for Changjiang Scholars and Innovative Research Team in University (No.IRT0944)
文摘A new butyrolactone derivative,namely butyrolactone Ⅷ （1）,and six known butyrolactones （2-7） were separated from the ethyl acetate （EtOAc） extract of the fermentation broth of a fungus,Aspergillus terreus MXH-23.The chemical structures of these metabolites were identified by analyzing their nuclear magnetic resonance （NMR） and mass spectrometry （MS）.Known butyrolactone derivatives contain an α,β-unsaturated γ-lactone ring with α-hydroxyl and y-benzyl,and butyrolactone Ⅷ （1） was the first butyrolactones contains α-benzyl and γ-hydroxyl on α,β-unsaturated lactone ring.All of the butyrolactone derivatives were tested for their anti-influenza （H 1N 1） effects.Derivatives 4 and 7 showed moderate antiviral activities while the newly-identified,derivative 1,did not.
文摘Aspergillus oryzae 3042 mycelium pellets were immobilized by crosslinking method with reagents of gelatin and formaldehyde. An orthogonal design table was used to determine the optimal immobilization conditions. The L-aminoacylase activity of immobilized mycelium pellets under optimal conditions was assayed. The results show that the L-aminoacylase activity was 1350 U·g^-1 and the activity retention rate was 83%. The properties of the immobilized preparation were studied. Compared with free pellets, the appropriate pH of reaction system changed from 7.0 to 8.0, temperature changed from 52℃ to 63℃, and the ranges of the optimum reaction conditions were all improved. The effect of Co^2＋ on immobilized mycelium pellets was investigated and the favorable concentration was determined. When the immobilized preparation was used for the resolution of D, L-alanine in a packed bed reactor with 10 mm in diameter, 200 mm in height, the operational stability was increased, and the half-life period was 53 d. Thermal stability analysis shows that the immobilized pellets were more stable than the free pellets.
文摘A strain HB-03 to produce alkaline extracellular lipase was isolated from oil-rich soil samples and identified as Aspergillus awamori. The growth conditions and nutritional factors for lipase production by strain HB-03 were optimized, and the maximum lipase production of （45.9±2.3） U/mL was obtained at 30 ℃ and pH 7.0 after 36 h using olive oil （1%） and sucrose （0.5%） as carbon sources and combination of peptone （2%）, yeast extract （0.5%） and ammonium sulfate （0.1%） as nitrogen sources. The lipase was purified to homogeneity with 10.6-fold, 18.84% yield and a specific activity of 1 862.2 U/mg using ammonium sulfate precipitation followed by SephadexG-75 gel filtration chromatography. The purified lipase with molecular mass of 68 ku was estimated by SDS-PAGE. The optimum pH and temperature for the purified lipase were found to be 8.5 and 40 ℃, respectively. The lipase kept more than 80% of activity in pH 7.0-10.0 and temperatures up to 45 ℃. The metal ions of Mn2＋, Ba2＋ significantly enhanced the lipase activity, whereas Cu2＋, Fe3＋ and Mg2＋ strongly reduced the lipase activity. The Km and Vmax values of the purified enzyme for p-nitrophenyl palmitate were 0.13 mrnol/L and 60.6 mmol/（L.min）, respectively. The results show that this novel lipase has potential industrial applications.
文摘Phosphorus （P） is a major growth-limiting nutrient, and unlike the case of nitrogen （N）, there is no large atmospheric source that can be made biologically available. Moreover, P governs crucial role in plant as it stimulates root development and growth, gives plant rapid and vigorous start leading to better tillering and essential for many metabolic processes for seed formation. Soil microbes play very important role in bio-weathering and biodegradation. The microorganisms produce low molecular mass organic acids, which attack the phosphate structure and transform phosphorus from non-utilizable to the utilizable for the plants form. The test of the relative efficiency of isolated strains is carried out by selecting the microorganisms that are capable of producing a halo/clear zone on a plate owing to the production of organic acids into the surrounding medium. It is a well-known fact that as the particle size of rock phosphate decreases, the microbe mediated solubilization of rock phosphate increases in soil. In the present investigation, microbial solubilization of nano rock phosphate （〈 100 nm） particles was studied. Experimental results revealed that Pseudomonas striata solubilized 11.45% of the total P after 24 h of incubation from nano rock phosphate particles while 28.95% and 21.19% of the total P was solubilized by Aspergillus niger （black pigmented） and Aspergillus niger （green pigmented）, respectively. It was also observed that Aspergillus niger has the higher ability to dissolve Udaipur rock phosphate than Pseudomonas striata.
文摘The objective of this study is to improve the production of L-DOPA from fungal source like Aspergillus terreus that can be further used to large-scale commercial production of this important drug from microbial sources. L-DOPA, a dopamine precursor that can pass the blood-brain barrier, is presently the drug of choice for Parkinson＇s disease. Microbial production and isolation of L-DOPA from natural sources is yet to be achieved an economical process. In this study, the mycelial pellets ofAspergillus terreus 104 were entrapped in 2% calcium alginate and were studied for their capacity for L-3, 4-dihydroxyphenylalanine production. Results showed that the immobilized pellets produced L-DOPA to the extent of 0.74 mg·G^-1 biomass while the free pellets produced 0.8 mg·G^-1 biomass. Further, storage of immobilized pellets for 96 h at 4 ℃ resulted in the reduction of the original L-DOPA producing activity of the gel beads only 40% and that of free pellets lost completely. In order to improve the production yield, further experiments were designed. It was found that L-DOPA production could be prolonged with repeated batch wise use of immobilized mycelial pellets in calcium alginate retaining 80% of their L-DOPA producing capacity for a period of 72 h while free pellets lost completely within 24 h. Results of this kind therefore is interesting and promising for commercial scale production of L-DOPA from microbial sources.
基金supported by the National Natural Science Foundation of China(Grant No.41173091)National Key Basic Research Program of China(Grant No.2013CB956702)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘We investigated the differences in the gene expression of carbonic anhydrase （CA） and cysteine synthase （CysM） between two weathering conditions, with either soluble potassium or insoluble potassium. We cultured a strain of A. niger by adopting a variant Czapek medium （using NazHPO4 as a substitute for KzHPO4） in two groups, Group A （containing silicate minerals bearing potassium but without KC1） and Group B （with KCI） . We extracted the mRNAs of CA and CysM from these two groups and performed real-time quantitative polymerase chain reactions （RT-qPCR）. We constructed relative standard curves by using glyceraldehyde-3-phosphate dehydrogenase （GAPDH） as the reference to confirm a consistent amplification effi- ciency of the target genes （CA and CysM） and the reference gene and quantified the gene expression of the targets in a relative manner. Our results showed that CA and CysM in Group A were upregulated for 1.7 times and 11.7 times, respectively, com- pared with those in Group B. Furthermore, we also analyzed some metabolic pathways and functions of the A. niger-induced weathering of potassium-bearing minerals, which involved the synthesizing of these two enzymes. Thus our work provides materials for further study of the roles of A. niger in the metabolic regulation during the weathering process of potassi- um-beating minerals.
