Pub Date : 2017-01-03DOI: 10.1080/10826068.2016.1275011
Y. Song, S. Baik
ABSTRACT A gene encoding cinnamoyl esterase (CE), which breaks down chlorogenic acid (ChA) into caffeic and quinic acids, was cloned from Lactobacillus helveticus KCCM 11223. The gene with an open reading frame of 759 nucleotides was expressed in Escherichia coli, which resulted in a 51.6-fold increase in specific activity compared to L. helveticus KCCM 11223. The recombinant CE exists as a monomeric enzyme having a molecular weight of 27.4 kDa. Although the highest activity was observed at pH 7, the enzyme showed stable activity at pH 4.0–10.0. Its optimum temperature was 65°C, and it also possessed a thermophilic activity: the half-life of CE was 24.4 min at 65°C. The half-life of CE was 145.5, 80.5, and 24.4 min at 60, 62, and 65°C, respectively. The Km and Vmax values for ChA were 0.153 mM and 559.6 µM/min, respectively. Moreover, the CE showed the highest substrate specificity with methyl caffeate among other methyl esters of hydroxycinnamic acids such as methyl ferulate, methyl sinapinate, methyl p-coumarate, and methyl caffeate. Ca2+, Cu2+, and Fe2+ significantly reduced the relative activity on ChA up to 70%. This is the first report on a thermostable CE from lactic acid bacteria that can be useful to hydrolyze ChA from plant cell walls.
{"title":"Molecular cloning, purification, and characterization of a novel thermostable cinnamoyl esterase from Lactobacillus helveticus KCCM 11223","authors":"Y. Song, S. Baik","doi":"10.1080/10826068.2016.1275011","DOIUrl":"https://doi.org/10.1080/10826068.2016.1275011","url":null,"abstract":"ABSTRACT A gene encoding cinnamoyl esterase (CE), which breaks down chlorogenic acid (ChA) into caffeic and quinic acids, was cloned from Lactobacillus helveticus KCCM 11223. The gene with an open reading frame of 759 nucleotides was expressed in Escherichia coli, which resulted in a 51.6-fold increase in specific activity compared to L. helveticus KCCM 11223. The recombinant CE exists as a monomeric enzyme having a molecular weight of 27.4 kDa. Although the highest activity was observed at pH 7, the enzyme showed stable activity at pH 4.0–10.0. Its optimum temperature was 65°C, and it also possessed a thermophilic activity: the half-life of CE was 24.4 min at 65°C. The half-life of CE was 145.5, 80.5, and 24.4 min at 60, 62, and 65°C, respectively. The Km and Vmax values for ChA were 0.153 mM and 559.6 µM/min, respectively. Moreover, the CE showed the highest substrate specificity with methyl caffeate among other methyl esters of hydroxycinnamic acids such as methyl ferulate, methyl sinapinate, methyl p-coumarate, and methyl caffeate. Ca2+, Cu2+, and Fe2+ significantly reduced the relative activity on ChA up to 70%. This is the first report on a thermostable CE from lactic acid bacteria that can be useful to hydrolyze ChA from plant cell walls.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76474474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-03DOI: 10.1080/10826068.2016.1275010
Dongsheng Xu, Aini Wan, Lin Peng, Yun Chen, Yang He, Jianfeng Yang, Jian Jin
ABSTRACT Hepatocyte growth factor (HGF) is a potent multifunctional cytokine that affects proliferation, migration, and morphogenesis of various cells. HGF is secreted as an inactive single-chain precursor protein and activated by the cleavage of serine proteases to form heterodimers. In our current study, the cleavage site of HGF was blocked by replaced Arg 494 of Glu (R494E) that resulted in the single-chain HGF (R494E) unable to be cleaved by serine proteases. We established Chinese hamster ovary (CHO) cells overexpressing HGF (R494E), the expression of HGF (R494E) achieved 12 mg/L and was similar to a previously reported study. The recombinant protein was then purified from culture medium using a two-step chromatographic procedure that resulted in about a 40% recovery rate. The purified HGF (R494E) was obtained as a single-chain active protein. It concluded that HGF (R494E) exhibited a biologically active protein and the overexpressing CHO cell line supplied sufficient material for future studies. The R494E replacement of the cleavage site would be beneficial to the utility of other similar therapeutic proteins.
{"title":"Production of human mutant biologically active hepatocyte growth factor in Chinese hamster ovary cells","authors":"Dongsheng Xu, Aini Wan, Lin Peng, Yun Chen, Yang He, Jianfeng Yang, Jian Jin","doi":"10.1080/10826068.2016.1275010","DOIUrl":"https://doi.org/10.1080/10826068.2016.1275010","url":null,"abstract":"ABSTRACT Hepatocyte growth factor (HGF) is a potent multifunctional cytokine that affects proliferation, migration, and morphogenesis of various cells. HGF is secreted as an inactive single-chain precursor protein and activated by the cleavage of serine proteases to form heterodimers. In our current study, the cleavage site of HGF was blocked by replaced Arg 494 of Glu (R494E) that resulted in the single-chain HGF (R494E) unable to be cleaved by serine proteases. We established Chinese hamster ovary (CHO) cells overexpressing HGF (R494E), the expression of HGF (R494E) achieved 12 mg/L and was similar to a previously reported study. The recombinant protein was then purified from culture medium using a two-step chromatographic procedure that resulted in about a 40% recovery rate. The purified HGF (R494E) was obtained as a single-chain active protein. It concluded that HGF (R494E) exhibited a biologically active protein and the overexpressing CHO cell line supplied sufficient material for future studies. The R494E replacement of the cleavage site would be beneficial to the utility of other similar therapeutic proteins.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85406513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-03DOI: 10.1080/10826068.2016.1275009
S. Birhade, Mukesh Pednekar, Shilpa Sagwal, A. Odaneth, A. Lali
ABSTRACT Controlled depolymerization of cellulose is essential for the production of valuable cellooligosaccharides and cellobiose from lignocellulosic biomass. However, enzymatic cellulose hydrolysis involves multiple synergistically acting enzymes, making difficult to control the depolymerization process and generate desired product. This work exploits the varying adsorption properties of the cellulase components to the cellulosic substrate and aims to control the enzyme activity. Cellulase adsorption was favored on pretreated cellulosic biomass as compared to synthetic cellulose. Preferential adsorption of exocellulases was observed over endocellulase, while β-glucosidases remained unadsorbed. Adsorbed enzyme fraction with bound exocellulases when used for hydrolysis generated cellobiose predominantly, while the unadsorbed enzymes in the liquid fraction produced cellooligosaccharides majorly, owing to its high endocellulases activity. Thus, the differential adsorption phenomenon of the cellulase components can be used for the controlling cellulose hydrolysis for the production of an array of sugars.
{"title":"Preparation of cellulase concoction using differential adsorption phenomenon","authors":"S. Birhade, Mukesh Pednekar, Shilpa Sagwal, A. Odaneth, A. Lali","doi":"10.1080/10826068.2016.1275009","DOIUrl":"https://doi.org/10.1080/10826068.2016.1275009","url":null,"abstract":"ABSTRACT Controlled depolymerization of cellulose is essential for the production of valuable cellooligosaccharides and cellobiose from lignocellulosic biomass. However, enzymatic cellulose hydrolysis involves multiple synergistically acting enzymes, making difficult to control the depolymerization process and generate desired product. This work exploits the varying adsorption properties of the cellulase components to the cellulosic substrate and aims to control the enzyme activity. Cellulase adsorption was favored on pretreated cellulosic biomass as compared to synthetic cellulose. Preferential adsorption of exocellulases was observed over endocellulase, while β-glucosidases remained unadsorbed. Adsorbed enzyme fraction with bound exocellulases when used for hydrolysis generated cellobiose predominantly, while the unadsorbed enzymes in the liquid fraction produced cellooligosaccharides majorly, owing to its high endocellulases activity. Thus, the differential adsorption phenomenon of the cellulase components can be used for the controlling cellulose hydrolysis for the production of an array of sugars.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85577383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-03DOI: 10.1080/10826068.2016.1275013
Alireza Moosavi Zenooz, F. Ashtiani, R. Ranjbar, F. Nikbakht, O. Bolouri
ABSTRACT Biodiesel production from microalgae feedstock should be performed after growth and harvesting of the cells, and the most feasible method for harvesting and dewatering of microalgae is flocculation. Flocculation modeling can be used for evaluation and prediction of its performance under different affective parameters. However, the modeling of flocculation in microalgae is not simple and has not performed yet, under all experimental conditions, mostly due to different behaviors of microalgae cells during the process under different flocculation conditions. In the current study, the modeling of microalgae flocculation is studied with different neural network architectures. Microalgae species, Chlorella sp., was flocculated with ferric chloride under different conditions and then the experimental data modeled using artificial neural network. Neural network architectures of multilayer perceptron (MLP) and radial basis function architectures, failed to predict the targets successfully, though, modeling was effective with ensemble architecture of MLP networks. Comparison between the performances of the ensemble and each individual network explains the ability of the ensemble architecture in microalgae flocculation modeling.
{"title":"Comparison of different artificial neural network architectures in modeling of Chlorella sp. flocculation","authors":"Alireza Moosavi Zenooz, F. Ashtiani, R. Ranjbar, F. Nikbakht, O. Bolouri","doi":"10.1080/10826068.2016.1275013","DOIUrl":"https://doi.org/10.1080/10826068.2016.1275013","url":null,"abstract":"ABSTRACT Biodiesel production from microalgae feedstock should be performed after growth and harvesting of the cells, and the most feasible method for harvesting and dewatering of microalgae is flocculation. Flocculation modeling can be used for evaluation and prediction of its performance under different affective parameters. However, the modeling of flocculation in microalgae is not simple and has not performed yet, under all experimental conditions, mostly due to different behaviors of microalgae cells during the process under different flocculation conditions. In the current study, the modeling of microalgae flocculation is studied with different neural network architectures. Microalgae species, Chlorella sp., was flocculated with ferric chloride under different conditions and then the experimental data modeled using artificial neural network. Neural network architectures of multilayer perceptron (MLP) and radial basis function architectures, failed to predict the targets successfully, though, modeling was effective with ensemble architecture of MLP networks. Comparison between the performances of the ensemble and each individual network explains the ability of the ensemble architecture in microalgae flocculation modeling.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75981706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-02DOI: 10.1080/10826068.2016.1155058
M. Imamura, N. Kato, Y. Iwamaru, S. Mohri, T. Yokoyama, Y. Murayama
ABSTRACT We previously showed that baculovirus-derived recombinant prion protein (Bac-PrP) can be converted to the misfolded infectious form (PrPSc) by protein misfolding cyclic amplification, an in vitro conversion technique. Bac-PrP, with post-translational modifications, would be useful for various applications such as using PrP as an immunogen for generating anti-PrP antibody, developing anti-prion drugs or diagnostic assays using in vitro conversion systems, and establishing an in vitro prion propagation model. For this purpose, highly purified Bac-PrP with in vitro conversion activity is necessary for use as a PrPC source, to minimize contamination. Furthermore, an exogenous affinity tag-free form is desirable to avoid potential steric interference by the affinity tags during the conversion process. In this study, we established purification methods for the untagged Bac-PrP under native conditions by combining exogenous double-affinity tags, namely, a polyhistidine-tag and a profinity eXact tag, with an octarepeat sequence of the N-terminal region of PrP, which has metal ion-binding affinity. The untagged Bac-PrP with near-homogeneity was obtained by three-step affinity purification, and it was shown that the final, purified Bac-PrP could convert to its pathogenic form. The presented purification procedure could be applied not only to PrP but also to other eukaryotic, recombinant proteins that require high purity and intact physiological activity.
{"title":"Multiple affinity purification of a baculovirus-derived recombinant prion protein with in vitro ability to convert to its pathogenic form","authors":"M. Imamura, N. Kato, Y. Iwamaru, S. Mohri, T. Yokoyama, Y. Murayama","doi":"10.1080/10826068.2016.1155058","DOIUrl":"https://doi.org/10.1080/10826068.2016.1155058","url":null,"abstract":"ABSTRACT We previously showed that baculovirus-derived recombinant prion protein (Bac-PrP) can be converted to the misfolded infectious form (PrPSc) by protein misfolding cyclic amplification, an in vitro conversion technique. Bac-PrP, with post-translational modifications, would be useful for various applications such as using PrP as an immunogen for generating anti-PrP antibody, developing anti-prion drugs or diagnostic assays using in vitro conversion systems, and establishing an in vitro prion propagation model. For this purpose, highly purified Bac-PrP with in vitro conversion activity is necessary for use as a PrPC source, to minimize contamination. Furthermore, an exogenous affinity tag-free form is desirable to avoid potential steric interference by the affinity tags during the conversion process. In this study, we established purification methods for the untagged Bac-PrP under native conditions by combining exogenous double-affinity tags, namely, a polyhistidine-tag and a profinity eXact tag, with an octarepeat sequence of the N-terminal region of PrP, which has metal ion-binding affinity. The untagged Bac-PrP with near-homogeneity was obtained by three-step affinity purification, and it was shown that the final, purified Bac-PrP could convert to its pathogenic form. The presented purification procedure could be applied not only to PrP but also to other eukaryotic, recombinant proteins that require high purity and intact physiological activity.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88279717","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-02DOI: 10.1080/10826068.2016.1155059
A. Kaur, A. Singh, A. Dua, R. Mahajan
ABSTRACT Simultaneous production of xylanase and pectinase by Bacillus pumilus AJK under submerged fermentation was investigated in this study. Under optimized conditions, it produced 315 ± 16 IU/mL acidic xylanase, 290 ± 20 IU/mL alkaline xylanase, and 88 ± 9 IU/mL pectinase. The production of xylano-pectinolytic enzymes was the highest after inoculating media (containing 2% each of wheat bran and Citrus limetta peel, 0.5% peptone, 10 mM MgSO4, pH 7.0) with 2% of 21-hr-old culture and incubated at 37°C for 60 hr at 200 rpm. Xylanase retained 100% activity from pH 6.0 to10.0 after 3 hr of incubation, while pectinase showed 100% stability from pH 6.0 to 9.0 even after 6 hr of incubation. Cost-effective and concurrent production of xylanase and pectinase by a bacterial isolate in the same production media suggests its potential for various biotechnological applications. This is the first report of simultaneous production of industrially important extracellular xylano-pectinolytic enzymes by B. pumilus.
{"title":"Cost-effective and concurrent production of industrially valuable xylano-pectinolytic enzymes by a bacterial isolate Bacillus pumilus AJK","authors":"A. Kaur, A. Singh, A. Dua, R. Mahajan","doi":"10.1080/10826068.2016.1155059","DOIUrl":"https://doi.org/10.1080/10826068.2016.1155059","url":null,"abstract":"ABSTRACT Simultaneous production of xylanase and pectinase by Bacillus pumilus AJK under submerged fermentation was investigated in this study. Under optimized conditions, it produced 315 ± 16 IU/mL acidic xylanase, 290 ± 20 IU/mL alkaline xylanase, and 88 ± 9 IU/mL pectinase. The production of xylano-pectinolytic enzymes was the highest after inoculating media (containing 2% each of wheat bran and Citrus limetta peel, 0.5% peptone, 10 mM MgSO4, pH 7.0) with 2% of 21-hr-old culture and incubated at 37°C for 60 hr at 200 rpm. Xylanase retained 100% activity from pH 6.0 to10.0 after 3 hr of incubation, while pectinase showed 100% stability from pH 6.0 to 9.0 even after 6 hr of incubation. Cost-effective and concurrent production of xylanase and pectinase by a bacterial isolate in the same production media suggests its potential for various biotechnological applications. This is the first report of simultaneous production of industrially important extracellular xylano-pectinolytic enzymes by B. pumilus.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91403800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-02DOI: 10.1080/10826068.2016.1163580
V. S. Avinash, Palna Dinesh Chauhan, Shraddha Y. Gaikwad, A. Pundle
ABSTRACT The production of 6-aminopenicillanic acid (6-APA) is a key step in the manufacture of semisynthetic antibiotics in the pharmaceutical industry. The penicillin G acylase from Escherichia coli has long been utilized for this purpose. However, the use of penicillin V acylases (PVA) presents some advantages including better stability and higher conversion rates. The industrial application of PVAs has so far been limited due to the nonavailability of suitable bacterial strains and cost issues. In this study, whole-cell immobilization of a recombinant PVA enzyme from Pectobacterium atrosepticum expressed in E. coli was performed. Membrane permeabilization with detergent was used to enhance the cell-bound PVA activity, and the cells were encapsulated in calcium alginate beads and cross-linked with glutaraldehyde. Optimization of parameters for the biotransformation by immobilized cells showed that full conversion of pen V to 6-APA could be achieved within 1 hr at pH 5.0 and 35°C, till 4% (w/v) concentration of the substrate. The beads could be stored for 28 days at 4°C with minimal loss in activity and were reusable up to 10 cycles with 1-hr hardening in CaCl2 between each cycle. The high enzyme productivity of the PVA enzyme system makes a promising case for its application for 6-APA production in the industry.
{"title":"Biotransformation of penicillin V to 6-aminopenicillanic acid using immobilized whole cells of E. coli expressing a highly active penicillin V acylase","authors":"V. S. Avinash, Palna Dinesh Chauhan, Shraddha Y. Gaikwad, A. Pundle","doi":"10.1080/10826068.2016.1163580","DOIUrl":"https://doi.org/10.1080/10826068.2016.1163580","url":null,"abstract":"ABSTRACT The production of 6-aminopenicillanic acid (6-APA) is a key step in the manufacture of semisynthetic antibiotics in the pharmaceutical industry. The penicillin G acylase from Escherichia coli has long been utilized for this purpose. However, the use of penicillin V acylases (PVA) presents some advantages including better stability and higher conversion rates. The industrial application of PVAs has so far been limited due to the nonavailability of suitable bacterial strains and cost issues. In this study, whole-cell immobilization of a recombinant PVA enzyme from Pectobacterium atrosepticum expressed in E. coli was performed. Membrane permeabilization with detergent was used to enhance the cell-bound PVA activity, and the cells were encapsulated in calcium alginate beads and cross-linked with glutaraldehyde. Optimization of parameters for the biotransformation by immobilized cells showed that full conversion of pen V to 6-APA could be achieved within 1 hr at pH 5.0 and 35°C, till 4% (w/v) concentration of the substrate. The beads could be stored for 28 days at 4°C with minimal loss in activity and were reusable up to 10 cycles with 1-hr hardening in CaCl2 between each cycle. The high enzyme productivity of the PVA enzyme system makes a promising case for its application for 6-APA production in the industry.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72699433","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-02DOI: 10.1080/10826068.2016.1155061
Xiaoqin Wang, Ying Jiang, Daode Hu
ABSTRACT The objective of the study was to optimize the ultrasonic-assisted extraction of curdione, furanodienone, curcumol, and germacrone from Curcuma phaeocaulis Valeton (Val.) and investigate the antiproliferative activity of the extract. Under the suitable high-performance liquid chromatography condition, the calibration curves for these four tested compounds showed high levels of linearity and the recoveries of these four compounds were between 97.9 and 104.3%. Response surface methodology (RSM) combining central composite design and desirability function (DF) was used to define optimal extraction parameters. The results of RSM and DF revealed that the optimum conditions were obtained as 8 mL g−1 for liquid–solid ratio, 70% ethanol concentration, and 20 min of ultrasonic time. It was found that the surface structures of the sonicated herbal materials were fluffy and irregular. The C. phaeocaulis Val. extract significantly inhibited the proliferation of RKO and HT-29 cells in vitro. The results reveal that the RSM can be effectively used for optimizing the ultrasonic-assisted extraction of bioactive components from C. phaeocaulis Val. for antiproliferative activity.
摘要:以姜黄为原料，优选超声辅助提取莪术中莪术酮、呋喃二烯酮、莪术酚和大黄酮的工艺条件，并考察其抗肿瘤活性。在合适的高效液相色谱条件下，4种化合物的标度曲线线性良好，加样回收率在97.9 ~ 104.3%之间。采用响应面法(RSM)结合中心组合设计和期望函数法(DF)确定最佳提取参数。RSM和DF结果表明，最佳条件为液固比为8 mL g−1，乙醇浓度为70%，超声时间为20 min。结果表明，经超声处理的药材表面结构呈蓬松状、不规则状。褐毛参提取物对RKO细胞和HT-29细胞的体外增殖有明显抑制作用。结果表明，RSM可有效地优化超声辅助提取法提取黄芪抗增殖活性成分的工艺条件。
{"title":"Antiproliferative activity of Curcuma phaeocaulis Valeton extract using ultrasonic assistance and response surface methodology","authors":"Xiaoqin Wang, Ying Jiang, Daode Hu","doi":"10.1080/10826068.2016.1155061","DOIUrl":"https://doi.org/10.1080/10826068.2016.1155061","url":null,"abstract":"ABSTRACT The objective of the study was to optimize the ultrasonic-assisted extraction of curdione, furanodienone, curcumol, and germacrone from Curcuma phaeocaulis Valeton (Val.) and investigate the antiproliferative activity of the extract. Under the suitable high-performance liquid chromatography condition, the calibration curves for these four tested compounds showed high levels of linearity and the recoveries of these four compounds were between 97.9 and 104.3%. Response surface methodology (RSM) combining central composite design and desirability function (DF) was used to define optimal extraction parameters. The results of RSM and DF revealed that the optimum conditions were obtained as 8 mL g−1 for liquid–solid ratio, 70% ethanol concentration, and 20 min of ultrasonic time. It was found that the surface structures of the sonicated herbal materials were fluffy and irregular. The C. phaeocaulis Val. extract significantly inhibited the proliferation of RKO and HT-29 cells in vitro. The results reveal that the RSM can be effectively used for optimizing the ultrasonic-assisted extraction of bioactive components from C. phaeocaulis Val. for antiproliferative activity.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86171327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-02DOI: 10.1080/10826068.2016.1155062
Sainan Liu, Wenzhu Tang, Fan Yang, J. Meng, Wenfu Chen, Xianzhen Li
ABSTRACT Biochar can enhance soil fertility to increase agricultural productivity, whereas its improvement in soil microbial activity is still unclear. In this article, the influence of biochar on the cell growth and the potassium-solubilizing activity of Bacillus mucilaginosus AS1153 was examined. The impact on cell growth is related to the biochar-derived feedstocks and the particle size of biochar. Both intrinsic features and inner component fraction can promote the cell growth of B. mucilaginosus AS1153. The potassium-solubilizing activity was increased by 80% when B. mucilaginosus was incubated in conjunction with the biochar derived from corn stover. The survival time of B. mucilaginosus also was prolonged by adsorption in biochar. The experimental results suggested that the biochar containing B. mucilaginosus could be used as a potential biofertilizer to sustain crop production.
{"title":"Influence of biochar application on potassium-solubilizing Bacillus mucilaginosus as potential biofertilizer","authors":"Sainan Liu, Wenzhu Tang, Fan Yang, J. Meng, Wenfu Chen, Xianzhen Li","doi":"10.1080/10826068.2016.1155062","DOIUrl":"https://doi.org/10.1080/10826068.2016.1155062","url":null,"abstract":"ABSTRACT Biochar can enhance soil fertility to increase agricultural productivity, whereas its improvement in soil microbial activity is still unclear. In this article, the influence of biochar on the cell growth and the potassium-solubilizing activity of Bacillus mucilaginosus AS1153 was examined. The impact on cell growth is related to the biochar-derived feedstocks and the particle size of biochar. Both intrinsic features and inner component fraction can promote the cell growth of B. mucilaginosus AS1153. The potassium-solubilizing activity was increased by 80% when B. mucilaginosus was incubated in conjunction with the biochar derived from corn stover. The survival time of B. mucilaginosus also was prolonged by adsorption in biochar. The experimental results suggested that the biochar containing B. mucilaginosus could be used as a potential biofertilizer to sustain crop production.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86853104","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-02DOI: 10.1080/10826068.2016.1163579
Narges Rahmanian, M. Bozorgmehr, monire torabi, A. Akbari, A. Zarnani
ABSTRACT Cell separation techniques play an indispensable part in numerous basic biological studies and even clinical settings. Although various cell isolation methods with diverse applications have been devised so far, not all of them have been able to gain widespread popularity among researchers and clinicians. There is not a single method known to be advantageous over all cell isolation techniques, and in fact, it is the researcher’s aim in performing a study that determines the most suitable method. A perfect method for one study might not be necessarily a proper choice for another and likewise, expensive and complex isolation methods might not always be the best choices. There are several criteria such as cell purity, viability, activation status, and frequency that need to be given serious thought before selecting an isolation technique. Moreover, time and cost are two of the key elements that should be taken into consideration before implementing a project. Hence, here we provide a succinct description of six more popular cell separation methods with respect to their principles, advantages, and disadvantages as well as their most common applications. We further provide several key features of each technique so that it helps the researchers to take the first step toward opting for the best method that fits well into their projects.
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