Ammonia（NH3） volatilization is one of the primary pathways of nitrogen（N） loss from soils after chemical fertilizer is applied, especially from the alkaline soils in Northern China, which results in lower efficien...Ammonia（NH3） volatilization is one of the primary pathways of nitrogen（N） loss from soils after chemical fertilizer is applied, especially from the alkaline soils in Northern China, which results in lower efficiency for chemical fertilizers. Therefore, we conducted an incubation experiment using an alkaline soil from Tianjin（p H 8.37–8.43） to evaluate the suppression effect of Trichoderma viride（T. viride） biofertilizer on NH3 volatilization, and compared the differences in microbial community structure among all samples. The results showed that viable T. viride biofertilizer（T） decreased NH3 volatilization by 42.21% compared with conventional fertilizer（（CK）, urea）, while nonviable T. viride biofertilizer（TS） decreased NH3 volatilization by 32.42%. NH3 volatilization was significantly higher in CK and sweet potato starch wastewater（SPSW） treatments during the peak period. T. viride biofertilizer also improved the transfer of ammonium from soil to sweet sorghum. Plant dry weights increased 91.23% and 61.08% for T and TS, respectively, compared to CK. Moreover, T. viride biofertilizer enhanced nitrification by increasing the abundance of ammonium-oxidizing archaea（AOA） and ammonium-oxidizing bacteria（AOB）. The results of high-throughput sequencing indicated that the microbial community structure and composition were significantly changed by the application of T. viride biofertilizer. This study demonstrated the immense potential of T. viride biofertilizer in reducing NH3 volatilization from alkaline soil and simultaneously improving the utilization of fertilizer N by sweet sorghum.展开更多
Biocontrol agents are safe and environmental friendly alternatives for pesticides in agriculture application. Trichoderma viride WEBL0703 performed a high level of antagonistic activity toward a broad spectrum of phyt...Biocontrol agents are safe and environmental friendly alternatives for pesticides in agriculture application. Trichoderma viride WEBL0703 performed a high level of antagonistic activity toward a broad spectrum of phytopathogens and was determined as a biocontrol agent, which was produced by solid state fermentation using grape marc and wine lees. The maximum yield of T. viride conidia was up to 6.65 × 10^9 CFU/g initial dry substrate （IDS） after 10 d fermentation. As important enzymes for protecting plants from disease, chitinase, β-glucanase, and pectinase yields were 47.8 U/g IDS, 8.32 U/g IDS and 9.83 U/g IDS, respectively. These results show that it is feasible to convert winery wastes to a value-added and environmental friendly biocontrol agent.展开更多
This paper reports the purification and characterization of kinetic parameters of cellulase produced from Trichoderma viride under still culture solid state fermentation technique using cheap and an easily available a...This paper reports the purification and characterization of kinetic parameters of cellulase produced from Trichoderma viride under still culture solid state fermentation technique using cheap and an easily available agricultural waste material, wheat straw as growth supported substrate. Trichoderma viride was cultured in fermentation medium of wheat straw under some previously optimized growth conditions and maximum activity of 398±2.43U/mL obtained after stipulated fermentation time period. Cellulase was purified 2.33 fold with specific activity of 105U/mg in comparison to crude enzyme extract using ammonium sulfate precipitation, dialysis and Sephadex-G-100 column chromatography. The enzyme was shown to have a relative low molecular weight of 58kDa by sodium dodecyl sulphate poly-acrylamide gel electrophoresis. The purified enzyme displayed 6.5 and 55oC as an optimum pH and temperature respectively. Using carboxymethyl cellulose as substrate, the enzyme showed maximum activity (Vmax) of 148U/mL with its corresponding KM value of 68μM. Among activators/inhibitors SDS, EDTA, and Hg2+ showed inhibitory effect on purified cellulase whereas, the enzyme activated by Co2+ and Mn2+ at a concentration of 1mM. The purified cellulase was compatible with four local detergent brands with up to 20 days of shelf life at room temperature suggesting its potential as a detergent additive for improved washing therefore, it is concluded that it may be potentially useful for industrial purposes especially for detergent and laundry industry.展开更多
[Objectives] This study aimed to establish HPLC fingerprint and conduct cluster analysis and principle component analysis for Citri Reticulatae Pericarpium Viride. [Methods] Using the HPLC method, the determination wa...[Objectives] This study aimed to establish HPLC fingerprint and conduct cluster analysis and principle component analysis for Citri Reticulatae Pericarpium Viride. [Methods] Using the HPLC method, the determination was performed on XSelect~® HSS T3-C_(18) column with mobile phase of acetonitrile-0.5% acetic acid solution(gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was 360 nm. The column temperature was 25℃. The sample size was 10 μL. With peak of hesperidin as the reference, HPLC fingerprints of 10 batches of Citri Reticulatae Pericarpium Viride were determined. The similarity of the 10 batches of samples was evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition) to determine the common peaks. Cluster analysis and principal component analysis were performed by using SPSS 17.0 statistical software. [Results] The HPLC fingerprints of the 10 batches of medicinal materials had total 11 common peaks, and the similarity was 0.919-1.000, indicating that the chemical composition of the 10 batches of medicinal materials was consistent. There were 11 common components in the 10 batches of medicinal materials, but their contents were different. When the Euclidean distance was 20, the 10 batches of samples were divided into two categories, S4 in the first category, and the others in the second one. When the Euclidean distance was 5, the second category could be further divided into two sub-categories, S1 and S10 in one sub-category, and S2, S3, S5, S6, S7, S8 and S9 in the other one. The principle component analysis showed that cumulative contribution rate of the two main component factors was 92.797%, and the comprehensive score of S7 was the highest with the best quality. [Conclusions] The results of HPLC fingerprinting, cluster analysis and principle component analysis can provide reference for the quality control of Citri Reticulatae Pericarpium Viride.展开更多
Two new tartrate derivative glucosides, coelovirin C (1) and D (2), were isolated from rhizomes of Coeloglossum viride ( L.) Hartm. var. bracteatum ( Willd.) Richter (Orchidaceae). Their structures were elucidated as ...Two new tartrate derivative glucosides, coelovirin C (1) and D (2), were isolated from rhizomes of Coeloglossum viride ( L.) Hartm. var. bracteatum ( Willd.) Richter (Orchidaceae). Their structures were elucidated as (2R, 3S)-2-b-D-glucopyranosyl-2-isobutyltartrate1-(4-b-D- glucopyranosyloxybenzyl) ester 1 and (2R, 3S)-2-b-D-glucopyranosyl-2-isobutyltartrate-4-(4-b-D- glucopyranosyloxybenzyl) ester 2 by means of chemical and spectroscopic methods.展开更多
In the present study, a co-culture technique was adopted with an aim to investigate a hyper production of exoglucanase, and β-glucosidase using cheap and easily available agro-industrial residue corn stover as growth...In the present study, a co-culture technique was adopted with an aim to investigate a hyper production of exoglucanase, and β-glucosidase using cheap and easily available agro-industrial residue corn stover as growth supporting substrate. Various physio-chemical and nutritional variables were optimized using classical and completely randomized designs for induced production of exoglucanase, and β-glucosidase from the co-culture of Trichoderma viride and Ganoderma lucidum in solid state fermentation (SSF). Analysis profile showed that when the conditions of the SSF medium containing 15 g corn stover substrate (50% w/w moisture) inoculated with 6 mL of inoculum were optimal, the maximum productions of exoglucanase (485 ± 6.5 U/mL) and β-glucosidase (255 ± 3.3 U/mL) were recorded after 5 days of incubation at pH 6 and 35°C.展开更多
Agro-industrial residues are primarily composed of complex polysaccharides that strengthen the microbial growth for the production of industrially important enzymes like cellulases. In the present study we aimed to ch...Agro-industrial residues are primarily composed of complex polysaccharides that strengthen the microbial growth for the production of industrially important enzymes like cellulases. In the present study we aimed to characterize the Exo 1, 4-β glucanase that was indigenously produced from Trichoderma viride MBL. T. viride MBL was cultured in the Solid-State medium of orange peel (50% w/w moisture) under optimized fermentation conditions and maximum activity of 412 ± 12 U/mL was recorded after 4th day of incubation at pH 5.5 and 30℃. Exo 1, 4-β glucanase was 4.17-fold purified with specific activity of 642 U/mg in comparison to the crude extract. To confirm its purity and molecular weight, sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS- PAGE) was performed. The enzyme was shown to have a molecular weight of 60 kDa with an optimum pH and temperature of 5 and 50℃, respectively. Lineweaver-Burk reciprocal plot revealed that the kinetic constants Km and Vmax of purified Exo 1, 4-β glucanase were 76 μM and 240 U/mL.展开更多
[Objective] To optimize solid state fermentation conditions of rapeseed meal and thus to reduce glucosinolates and neutral detergent fibers by mixed cultures of Aspergillus oryzae and Tnchoderrna viride. [ Method ] Th...[Objective] To optimize solid state fermentation conditions of rapeseed meal and thus to reduce glucosinolates and neutral detergent fibers by mixed cultures of Aspergillus oryzae and Tnchoderrna viride. [ Method ] The optimal fermentation conditions were determined by single factor test and orthogonal design. [ Result J The optimum fermentation conditions are as following: inoculum weight ratio (Aspergillus oryzae vs Trichoderma vinde), 1:1 ; inoculum size, 30% ; water content, 40% ; fermentation time, 96 h; and fermentation temperature, 30℃. Under these conditions, glucosinolates were reduced by 90.71% and neutral detergent fibers were degraded by 20.65%. [ Condusion] In laboratory, solid state fermentation with Aspergillus oryzae and Trichoderrna viride can be used to produce high-quality rapeseed meal.展开更多
Today, demand exists for cost-effective production of industrially important enzymes from entire scientific sectors. By keeping in mind the extensive industrial applications of cellulase, this study was performed to i...Today, demand exists for cost-effective production of industrially important enzymes from entire scientific sectors. By keeping in mind the extensive industrial applications of cellulase, this study was performed to immobilize the indigenous enzyme produced from Trichoderma viride under pre-optimized SSF of an agricultural waste material, wheat straw. To enhance the bio-catalytic and tolerance properties of the present enzyme gel matrix immobilization engineering was applied. Previously, 2.33~fold purified novel cellulase was immobilized in to a xerogel matrix of TMOS and PTMS. FTIR spectroscopy confirmed the successful immobilization of cellulase. The free and immobilized cellulase was characterized and stability profile showed that after 24 h incubation, immobilization enhanced the thermo-stability up to 75% against 80℃ as compare to the free enzyme. Xerogel matrix immobilization enhanced the catalytic efficiency of entrapped enzyme than that of the free cellulase. Among activators/inhibitors SDS, EDTA, and Hg2+ showed inhibitory effect while, gel matrix immobilization enhanced 80% tolerance capacity of the cellulase against inactivating agents.展开更多
Two new isobutyltartrate monoesters, coelovirin A (1) and B (2), were isolated from the rhizomes of Coeloglossum viride (L.) Hartm. var. bracteatum (Willd.) Richter (Orchidaceae). Their structures were elucidated as ...Two new isobutyltartrate monoesters, coelovirin A (1) and B (2), were isolated from the rhizomes of Coeloglossum viride (L.) Hartm. var. bracteatum (Willd.) Richter (Orchidaceae). Their structures were elucidated as (2R, 3S)-2-isobutyltartrate1-(4-b-D-glucopyranosyloxybenzyl) ester 1 and (2R, 3S)-2-isobutyltartrate4-(4-b-D-glucopyranosyloxybenzyl) ester 2 on the basis of physical constants and spectroscopic methods including 2D NMR techniques.展开更多
In the present study, solid state fermentation was carried out using orange peel waste to produce β-glucosidase from Trichoderma viride. A locally isolated fungal strain T. viride was cultured in the solid state medi...In the present study, solid state fermentation was carried out using orange peel waste to produce β-glucosidase from Trichoderma viride. A locally isolated fungal strain T. viride was cultured in the solid state medium of orange peel (50% w/w moisture) under optimized fermentation conditions and maximum activity of 515 ± 12.4 U/mL was recorded after 4th day of incubation at pH 5.5 and 30℃. Indigenously produced β-glucosidase was subjected to the ammonium sulfate precipitation and Sephadex-G-100 gel filtration chromatography. In comparison to the crude extract β-glucosidase was 5.1-fold purified with specific activity of 758 U/mg. The enzyme was shown to have a relative molecular weight of 62 kDa as evidenced by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The purified β-glucosidase displayed 6 and 60℃ as an optimum pH and temperature respectively.展开更多
基金supported by the National Science Fund Projects (Nos. 41371266 and 31670507)Innovation in Cross-functional Team Program of the Chinese Academy of Sciences (No. 2015)+1 种基金the Key Research Program of Chinese Academy of Sciences (No. ZDRW-ZS-2016-5)the Key State Science and Technology Program of China (No. 2015ZX07206-006)
文摘Ammonia（NH3） volatilization is one of the primary pathways of nitrogen（N） loss from soils after chemical fertilizer is applied, especially from the alkaline soils in Northern China, which results in lower efficiency for chemical fertilizers. Therefore, we conducted an incubation experiment using an alkaline soil from Tianjin（p H 8.37–8.43） to evaluate the suppression effect of Trichoderma viride（T. viride） biofertilizer on NH3 volatilization, and compared the differences in microbial community structure among all samples. The results showed that viable T. viride biofertilizer（T） decreased NH3 volatilization by 42.21% compared with conventional fertilizer（（CK）, urea）, while nonviable T. viride biofertilizer（TS） decreased NH3 volatilization by 32.42%. NH3 volatilization was significantly higher in CK and sweet potato starch wastewater（SPSW） treatments during the peak period. T. viride biofertilizer also improved the transfer of ammonium from soil to sweet sorghum. Plant dry weights increased 91.23% and 61.08% for T and TS, respectively, compared to CK. Moreover, T. viride biofertilizer enhanced nitrification by increasing the abundance of ammonium-oxidizing archaea（AOA） and ammonium-oxidizing bacteria（AOB）. The results of high-throughput sequencing indicated that the microbial community structure and composition were significantly changed by the application of T. viride biofertilizer. This study demonstrated the immense potential of T. viride biofertilizer in reducing NH3 volatilization from alkaline soil and simultaneously improving the utilization of fertilizer N by sweet sorghum.
基金supported by the Australian Research Council International Linkage Fellowship(No.LX0560210)the National Natural Science Foundation of China(No.30600082).
文摘Biocontrol agents are safe and environmental friendly alternatives for pesticides in agriculture application. Trichoderma viride WEBL0703 performed a high level of antagonistic activity toward a broad spectrum of phytopathogens and was determined as a biocontrol agent, which was produced by solid state fermentation using grape marc and wine lees. The maximum yield of T. viride conidia was up to 6.65 × 10^9 CFU/g initial dry substrate （IDS） after 10 d fermentation. As important enzymes for protecting plants from disease, chitinase, β-glucanase, and pectinase yields were 47.8 U/g IDS, 8.32 U/g IDS and 9.83 U/g IDS, respectively. These results show that it is feasible to convert winery wastes to a value-added and environmental friendly biocontrol agent.
文摘This paper reports the purification and characterization of kinetic parameters of cellulase produced from Trichoderma viride under still culture solid state fermentation technique using cheap and an easily available agricultural waste material, wheat straw as growth supported substrate. Trichoderma viride was cultured in fermentation medium of wheat straw under some previously optimized growth conditions and maximum activity of 398±2.43U/mL obtained after stipulated fermentation time period. Cellulase was purified 2.33 fold with specific activity of 105U/mg in comparison to crude enzyme extract using ammonium sulfate precipitation, dialysis and Sephadex-G-100 column chromatography. The enzyme was shown to have a relative low molecular weight of 58kDa by sodium dodecyl sulphate poly-acrylamide gel electrophoresis. The purified enzyme displayed 6.5 and 55oC as an optimum pH and temperature respectively. Using carboxymethyl cellulose as substrate, the enzyme showed maximum activity (Vmax) of 148U/mL with its corresponding KM value of 68μM. Among activators/inhibitors SDS, EDTA, and Hg2+ showed inhibitory effect on purified cellulase whereas, the enzyme activated by Co2+ and Mn2+ at a concentration of 1mM. The purified cellulase was compatible with four local detergent brands with up to 20 days of shelf life at room temperature suggesting its potential as a detergent additive for improved washing therefore, it is concluded that it may be potentially useful for industrial purposes especially for detergent and laundry industry.
基金Supported by National Natural Science Foundation of China(81603251)Key Research and Development Plan of Shanxi Province(201603D3113021)Project of Collaborative Innovation Center for the Comprehensive Development and Utilization of Medicinal Herbs in Shanxi Province(2017-JYXT-05)
文摘[Objectives] This study aimed to establish HPLC fingerprint and conduct cluster analysis and principle component analysis for Citri Reticulatae Pericarpium Viride. [Methods] Using the HPLC method, the determination was performed on XSelect~® HSS T3-C_(18) column with mobile phase of acetonitrile-0.5% acetic acid solution(gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was 360 nm. The column temperature was 25℃. The sample size was 10 μL. With peak of hesperidin as the reference, HPLC fingerprints of 10 batches of Citri Reticulatae Pericarpium Viride were determined. The similarity of the 10 batches of samples was evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition) to determine the common peaks. Cluster analysis and principal component analysis were performed by using SPSS 17.0 statistical software. [Results] The HPLC fingerprints of the 10 batches of medicinal materials had total 11 common peaks, and the similarity was 0.919-1.000, indicating that the chemical composition of the 10 batches of medicinal materials was consistent. There were 11 common components in the 10 batches of medicinal materials, but their contents were different. When the Euclidean distance was 20, the 10 batches of samples were divided into two categories, S4 in the first category, and the others in the second one. When the Euclidean distance was 5, the second category could be further divided into two sub-categories, S1 and S10 in one sub-category, and S2, S3, S5, S6, S7, S8 and S9 in the other one. The principle component analysis showed that cumulative contribution rate of the two main component factors was 92.797%, and the comprehensive score of S7 was the highest with the best quality. [Conclusions] The results of HPLC fingerprinting, cluster analysis and principle component analysis can provide reference for the quality control of Citri Reticulatae Pericarpium Viride.
文摘Two new tartrate derivative glucosides, coelovirin C (1) and D (2), were isolated from rhizomes of Coeloglossum viride ( L.) Hartm. var. bracteatum ( Willd.) Richter (Orchidaceae). Their structures were elucidated as (2R, 3S)-2-b-D-glucopyranosyl-2-isobutyltartrate1-(4-b-D- glucopyranosyloxybenzyl) ester 1 and (2R, 3S)-2-b-D-glucopyranosyl-2-isobutyltartrate-4-(4-b-D- glucopyranosyloxybenzyl) ester 2 by means of chemical and spectroscopic methods.
文摘In the present study, a co-culture technique was adopted with an aim to investigate a hyper production of exoglucanase, and β-glucosidase using cheap and easily available agro-industrial residue corn stover as growth supporting substrate. Various physio-chemical and nutritional variables were optimized using classical and completely randomized designs for induced production of exoglucanase, and β-glucosidase from the co-culture of Trichoderma viride and Ganoderma lucidum in solid state fermentation (SSF). Analysis profile showed that when the conditions of the SSF medium containing 15 g corn stover substrate (50% w/w moisture) inoculated with 6 mL of inoculum were optimal, the maximum productions of exoglucanase (485 ± 6.5 U/mL) and β-glucosidase (255 ± 3.3 U/mL) were recorded after 5 days of incubation at pH 6 and 35°C.
文摘Agro-industrial residues are primarily composed of complex polysaccharides that strengthen the microbial growth for the production of industrially important enzymes like cellulases. In the present study we aimed to characterize the Exo 1, 4-β glucanase that was indigenously produced from Trichoderma viride MBL. T. viride MBL was cultured in the Solid-State medium of orange peel (50% w/w moisture) under optimized fermentation conditions and maximum activity of 412 ± 12 U/mL was recorded after 4th day of incubation at pH 5.5 and 30℃. Exo 1, 4-β glucanase was 4.17-fold purified with specific activity of 642 U/mg in comparison to the crude extract. To confirm its purity and molecular weight, sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS- PAGE) was performed. The enzyme was shown to have a molecular weight of 60 kDa with an optimum pH and temperature of 5 and 50℃, respectively. Lineweaver-Burk reciprocal plot revealed that the kinetic constants Km and Vmax of purified Exo 1, 4-β glucanase were 76 μM and 240 U/mL.
基金funded by the grants from National High Technology Research and Development Program of China (2009A-A02Z210)Key Technology R&D Program of Qingdao City,China (07-2-1-25-nsh)
文摘[Objective] To optimize solid state fermentation conditions of rapeseed meal and thus to reduce glucosinolates and neutral detergent fibers by mixed cultures of Aspergillus oryzae and Tnchoderrna viride. [ Method ] The optimal fermentation conditions were determined by single factor test and orthogonal design. [ Result J The optimum fermentation conditions are as following: inoculum weight ratio (Aspergillus oryzae vs Trichoderma vinde), 1:1 ; inoculum size, 30% ; water content, 40% ; fermentation time, 96 h; and fermentation temperature, 30℃. Under these conditions, glucosinolates were reduced by 90.71% and neutral detergent fibers were degraded by 20.65%. [ Condusion] In laboratory, solid state fermentation with Aspergillus oryzae and Trichoderrna viride can be used to produce high-quality rapeseed meal.
文摘Today, demand exists for cost-effective production of industrially important enzymes from entire scientific sectors. By keeping in mind the extensive industrial applications of cellulase, this study was performed to immobilize the indigenous enzyme produced from Trichoderma viride under pre-optimized SSF of an agricultural waste material, wheat straw. To enhance the bio-catalytic and tolerance properties of the present enzyme gel matrix immobilization engineering was applied. Previously, 2.33~fold purified novel cellulase was immobilized in to a xerogel matrix of TMOS and PTMS. FTIR spectroscopy confirmed the successful immobilization of cellulase. The free and immobilized cellulase was characterized and stability profile showed that after 24 h incubation, immobilization enhanced the thermo-stability up to 75% against 80℃ as compare to the free enzyme. Xerogel matrix immobilization enhanced the catalytic efficiency of entrapped enzyme than that of the free cellulase. Among activators/inhibitors SDS, EDTA, and Hg2+ showed inhibitory effect while, gel matrix immobilization enhanced 80% tolerance capacity of the cellulase against inactivating agents.
文摘Two new isobutyltartrate monoesters, coelovirin A (1) and B (2), were isolated from the rhizomes of Coeloglossum viride (L.) Hartm. var. bracteatum (Willd.) Richter (Orchidaceae). Their structures were elucidated as (2R, 3S)-2-isobutyltartrate1-(4-b-D-glucopyranosyloxybenzyl) ester 1 and (2R, 3S)-2-isobutyltartrate4-(4-b-D-glucopyranosyloxybenzyl) ester 2 on the basis of physical constants and spectroscopic methods including 2D NMR techniques.
文摘In the present study, solid state fermentation was carried out using orange peel waste to produce β-glucosidase from Trichoderma viride. A locally isolated fungal strain T. viride was cultured in the solid state medium of orange peel (50% w/w moisture) under optimized fermentation conditions and maximum activity of 515 ± 12.4 U/mL was recorded after 4th day of incubation at pH 5.5 and 30℃. Indigenously produced β-glucosidase was subjected to the ammonium sulfate precipitation and Sephadex-G-100 gel filtration chromatography. In comparison to the crude extract β-glucosidase was 5.1-fold purified with specific activity of 758 U/mg. The enzyme was shown to have a relative molecular weight of 62 kDa as evidenced by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The purified β-glucosidase displayed 6 and 60℃ as an optimum pH and temperature respectively.
