Pub Date : 2017-03-16DOI: 10.1080/10826068.2016.1244682
Eun Jae Kim, J. Lee, S. G. Lee, S. Han
ABSTRACT To survive in a subzero environment, polar organisms produce ice-binding proteins (IBPs). These IBPs prevent the formation of large intracellular ice crystals, which may be fatal to the organism. Recently, a recombinant FfIBP (an IBP from Flavobacterium frigoris PS1) was cloned and produced in Pichia pastoris using fed-batch fermentation with methanol feeding. In this study, we demonstrate that FfIBP produced by P. pastoris has a glycosylation site, which diminishes the thermal hysteresis activity of FfIBP. The FfIBP expressed by P. pastoris exhibited a doublet on SDS-PAGE. The results of a glycosidase reaction suggested that FfIBP possesses complex N-linked oligosaccharides. These results indicate that the residues of the glycosylated site could disturb the binding of FfIBP to ice molecules. The findings of this study could be utilized to produce highly active antifreeze proteins on a large scale.
{"title":"Improving thermal hysteresis activity of antifreeze protein from recombinant Pichia pastoris by removal of N-glycosylation","authors":"Eun Jae Kim, J. Lee, S. G. Lee, S. Han","doi":"10.1080/10826068.2016.1244682","DOIUrl":"https://doi.org/10.1080/10826068.2016.1244682","url":null,"abstract":"ABSTRACT To survive in a subzero environment, polar organisms produce ice-binding proteins (IBPs). These IBPs prevent the formation of large intracellular ice crystals, which may be fatal to the organism. Recently, a recombinant FfIBP (an IBP from Flavobacterium frigoris PS1) was cloned and produced in Pichia pastoris using fed-batch fermentation with methanol feeding. In this study, we demonstrate that FfIBP produced by P. pastoris has a glycosylation site, which diminishes the thermal hysteresis activity of FfIBP. The FfIBP expressed by P. pastoris exhibited a doublet on SDS-PAGE. The results of a glycosidase reaction suggested that FfIBP possesses complex N-linked oligosaccharides. These results indicate that the residues of the glycosylated site could disturb the binding of FfIBP to ice molecules. The findings of this study could be utilized to produce highly active antifreeze proteins on a large scale.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"1 1","pages":"299 - 304"},"PeriodicalIF":0.0,"publicationDate":"2017-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84005358","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 : 2017-03-16DOI: 10.1080/10826068.2016.1244681
Ahmed Abd Elrazak, A. Ward, J. Glassey
ABSTRACT The increasing demand of omega-3 in the market and the challenges facing its conventional supplies led to an increasing interest to microbial omega-3 sources. This research concentrates on the statistical role of some metal ions on the biosynthesis and productivity of eicosapentaenoic acid (essential omega-3 element) in bacterial isolate, Shewanella 717. A Plackett–Burman design was applied to screen the main effect of all metal salts entrenched in the artificial sea water medium components. Four salts, in particular, in addition to the interaction among them were highlighted as having a statistically significant effect upon the growth and/or eicosapentaenoic acid production. A subsequent central composite design was performed to determine the exact optimum concentration of each of the chosen variables which was found to be 2.5, 1.8, 1.2, and 23 g/l, for Na2HPO4, MgSO4, KCl, and NaCl, respectively. All the experiments were performed with the minimal amount of carbon and nitrogen to eliminate any potential masking effect. A bioreactor batch run was operated and the ion uptake was monitored, using EDAX® electron microscopy, concluding that the process of microbial omega-3 production could be a phosphate-limited process. Optimizing the concentration of the tested metal ions led to a remarkable increase in the omega-3 productivity resulted in a 30, 9, and 10 times increase in yield, concentration, and percentage to the total fatty acids, respectively, even though the carbon and nitrogen were kept constant all over the research work.
{"title":"Effect of inorganic salts on bacterial omega-3 PUFA production","authors":"Ahmed Abd Elrazak, A. Ward, J. Glassey","doi":"10.1080/10826068.2016.1244681","DOIUrl":"https://doi.org/10.1080/10826068.2016.1244681","url":null,"abstract":"ABSTRACT The increasing demand of omega-3 in the market and the challenges facing its conventional supplies led to an increasing interest to microbial omega-3 sources. This research concentrates on the statistical role of some metal ions on the biosynthesis and productivity of eicosapentaenoic acid (essential omega-3 element) in bacterial isolate, Shewanella 717. A Plackett–Burman design was applied to screen the main effect of all metal salts entrenched in the artificial sea water medium components. Four salts, in particular, in addition to the interaction among them were highlighted as having a statistically significant effect upon the growth and/or eicosapentaenoic acid production. A subsequent central composite design was performed to determine the exact optimum concentration of each of the chosen variables which was found to be 2.5, 1.8, 1.2, and 23 g/l, for Na2HPO4, MgSO4, KCl, and NaCl, respectively. All the experiments were performed with the minimal amount of carbon and nitrogen to eliminate any potential masking effect. A bioreactor batch run was operated and the ion uptake was monitored, using EDAX® electron microscopy, concluding that the process of microbial omega-3 production could be a phosphate-limited process. Optimizing the concentration of the tested metal ions led to a remarkable increase in the omega-3 productivity resulted in a 30, 9, and 10 times increase in yield, concentration, and percentage to the total fatty acids, respectively, even though the carbon and nitrogen were kept constant all over the research work.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"5 1","pages":"291 - 298"},"PeriodicalIF":0.0,"publicationDate":"2017-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84866235","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 : 2017-03-16DOI: 10.1080/10826068.2016.1244684
Yanyang Xu, Yang Qiu, Hui Ren, D. Ju, Hong-Lei Jia
ABSTRACT Aronia melanocarpa berries are abundant in polyphenolic compounds. After juice production, the pomace of pressed berries still contains a substantial amount of polyphenolic compounds. For efficient utilization of A. melanocarpa berries and the enhancement of polyphenolic compound yields in Aronia melanocarpa pomace (AMP), total phenolics (TP) and total flavonoids (TF) from AMP were extracted, using ultrasound-assisted aqueous two-phase system (UAE-ATPS) extraction method. First, the influences of ammonium sulfate concentration, ethanol–water ratio, ultrasonic time, and ultrasonic power on TP and TF yields were investigated. On this basis, process variables such as ammonium sulfate concentration (0.30–0.35 g mL−1), ethanol–water ratio (0.6–0.8), ultrasonic time (40–60 min), and ultrasonic power (175–225 W) were further optimized by implementing Box–Benhnken design with response surface methodology. The experimental results showed that optimal extraction conditions of TP from AMP were as follows: ammonium sulfate concentration of 0.324 g mL−1, ethanol–water ratio of 0.69, ultrasonic time of 52 min, and ultrasonic power of 200 W. Meanwhile, ammonium sulfate concentration of 0.320 g mL−1, ethanol–water ratio of 0.71, ultrasonic time of 50 min, and ultrasonic power of 200 W were determined as optimum extraction conditions of TF in AMP. Experimental validation was performed, where TP and TF yields reached 68.15 ± 1.04 and 11.67 ± 0.63 mg g−1, respectively. Close agreement was found between experimental and predicted values. Overall, the present results demonstrated that ultrasound-assisted aqueous two-phase system extraction method was successfully used to extract total phenolics and flavonoids in A. melanocarpa pomace.
黑桫椤果实含有丰富的多酚类化合物。榨汁后，榨出的果渣仍然含有大量的多酚类化合物。为有效利用黑果莓果实，提高黑果莓果渣(AMP)中多酚类化合物的得率，采用超声辅助双水相萃取法(UAE-ATPS)提取AMP中的总酚类物质(TP)和总黄酮(TF)。首先，考察了硫酸铵浓度、乙醇水比、超声时间和超声功率对TP和TF产率的影响。在此基础上，采用Box-Benhnken设计，结合响应面法对硫酸铵浓度(0.30 ~ 0.35 g mL−1)、乙醇水比(0.6 ~ 0.8)、超声时间(40 ~ 60 min)、超声功率(175 ~ 225 W)等工艺参数进行进一步优化。实验结果表明，从AMP中提取TP的最佳工艺条件为:硫酸铵浓度0.324 g mL−1，乙醇-水比0.69，超声时间52 min，超声功率200 W。同时，确定了硫酸铵浓度为0.320 g mL−1、乙醇水比为0.71、超声时间为50 min、超声功率为200 W为提取AMP中总皂苷的最佳条件。实验验证，总皂苷和总皂苷得率分别为68.15±1.04和11.67±0.63 mg g−1。实验值与预测值非常吻合。综上所述，超声辅助双水相萃取法可有效提取黑桃果渣中的总酚类物质和总黄酮。
{"title":"Optimization of ultrasound-assisted aqueous two-phase system extraction of polyphenolic compounds from Aronia melanocarpa pomace by response surface methodology","authors":"Yanyang Xu, Yang Qiu, Hui Ren, D. Ju, Hong-Lei Jia","doi":"10.1080/10826068.2016.1244684","DOIUrl":"https://doi.org/10.1080/10826068.2016.1244684","url":null,"abstract":"ABSTRACT Aronia melanocarpa berries are abundant in polyphenolic compounds. After juice production, the pomace of pressed berries still contains a substantial amount of polyphenolic compounds. For efficient utilization of A. melanocarpa berries and the enhancement of polyphenolic compound yields in Aronia melanocarpa pomace (AMP), total phenolics (TP) and total flavonoids (TF) from AMP were extracted, using ultrasound-assisted aqueous two-phase system (UAE-ATPS) extraction method. First, the influences of ammonium sulfate concentration, ethanol–water ratio, ultrasonic time, and ultrasonic power on TP and TF yields were investigated. On this basis, process variables such as ammonium sulfate concentration (0.30–0.35 g mL−1), ethanol–water ratio (0.6–0.8), ultrasonic time (40–60 min), and ultrasonic power (175–225 W) were further optimized by implementing Box–Benhnken design with response surface methodology. The experimental results showed that optimal extraction conditions of TP from AMP were as follows: ammonium sulfate concentration of 0.324 g mL−1, ethanol–water ratio of 0.69, ultrasonic time of 52 min, and ultrasonic power of 200 W. Meanwhile, ammonium sulfate concentration of 0.320 g mL−1, ethanol–water ratio of 0.71, ultrasonic time of 50 min, and ultrasonic power of 200 W were determined as optimum extraction conditions of TF in AMP. Experimental validation was performed, where TP and TF yields reached 68.15 ± 1.04 and 11.67 ± 0.63 mg g−1, respectively. Close agreement was found between experimental and predicted values. Overall, the present results demonstrated that ultrasound-assisted aqueous two-phase system extraction method was successfully used to extract total phenolics and flavonoids in A. melanocarpa pomace.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"48 1","pages":"312 - 321"},"PeriodicalIF":0.0,"publicationDate":"2017-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73064882","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 : 2017-03-16DOI: 10.1080/10826068.2016.1244683
Aleksandra Margetić, Z. Vujčić
ABSTRACT Yeast Saccharomyces cerevisiae is the most significant source of enzyme invertase. It is mainly used in the food industry as a soluble or immobilized enzyme. The greatest amount of invertase is located in the periplasmic space in yeast. In this work, it was isolated into two forms of enzyme from yeast S. cerevisiae cell, soluble and cell wall invertase (CWI). Both forms of enzyme showed same temperature optimum (60°C), similar pH optimum, and kinetic parameters. The significant difference between these biocatalysts was observed in their thermal stability, stability in urea and methanol solution. At 60°C, CWI had 1.7 times longer half-life than soluble enzyme, while at 70°C CWI showed 8.7 times longer half-life than soluble enzyme. After 2-hr of incubation in 8 M urea solution, soluble invertase and CWI retained 10 and 60% of its initial activity, respectively. During 22 hr of incubation of both enzymes in 30 and 40% methanol, soluble invertase was completely inactivated, while CWI changed its activity within the experimental error. Therefore, soluble invertase and CWI have not shown any substantial difference, but CWI showed better thermal stability and stability in some of the typical protein-denaturing agents.
{"title":"Comparative study of stability of soluble and cell wall invertase from Saccharomyces cerevisiae","authors":"Aleksandra Margetić, Z. Vujčić","doi":"10.1080/10826068.2016.1244683","DOIUrl":"https://doi.org/10.1080/10826068.2016.1244683","url":null,"abstract":"ABSTRACT Yeast Saccharomyces cerevisiae is the most significant source of enzyme invertase. It is mainly used in the food industry as a soluble or immobilized enzyme. The greatest amount of invertase is located in the periplasmic space in yeast. In this work, it was isolated into two forms of enzyme from yeast S. cerevisiae cell, soluble and cell wall invertase (CWI). Both forms of enzyme showed same temperature optimum (60°C), similar pH optimum, and kinetic parameters. The significant difference between these biocatalysts was observed in their thermal stability, stability in urea and methanol solution. At 60°C, CWI had 1.7 times longer half-life than soluble enzyme, while at 70°C CWI showed 8.7 times longer half-life than soluble enzyme. After 2-hr of incubation in 8 M urea solution, soluble invertase and CWI retained 10 and 60% of its initial activity, respectively. During 22 hr of incubation of both enzymes in 30 and 40% methanol, soluble invertase was completely inactivated, while CWI changed its activity within the experimental error. Therefore, soluble invertase and CWI have not shown any substantial difference, but CWI showed better thermal stability and stability in some of the typical protein-denaturing agents.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"24 1","pages":"305 - 311"},"PeriodicalIF":0.0,"publicationDate":"2017-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81503579","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 : 2017-03-16DOI: 10.1080/10826068.2016.1207082
Rengarajan Saravanan, Shajahan Shubethar, S. Narayanan, M. Jain, S. Lade, D. Jadhav, Palamalai Maheswaran, Uday K Avalakki, A. Dubey
ABSTRACT Galactooligosaccharides (GOS) are nondigestible dietary fibers which have a beneficial effect on human health by promoting the growth of probiotic bacteria in the gut. In addition, other health benefits have been reported from oligosaccharides consumption such as stimulation of intestinal mobility, colon cancer prevention, mineral absorption as well as protection against certain pathogenic bacterial infections. The goal of this research was to develop an efficient biotransformation system using a consortium of microbes for the production of ≥85% pure GOS and reusing the cell biomass in repeated cycles of biotransformation. Production of GOS by lactose transgalactosylation using whole cells of Sporobolomyces singularis MTCC 5491 as a source of β-galactosidase and monosaccharides utilization by yeast isolate (NUTIDY007) were studied. For increasing the purity of GOS, growth and bioconversion parameters on the transgalactosylation by the whole cells were investigated. Further, continuous production of GOS was studied in a reactor with microfiltration membrane system. A maximum GOS purity of 42% was achieved using single culture of S. singularis. Under optimized conditions, single culture of S. singularis produced a maximum of 56% pure GOS. Addition of second culture to the reaction mixture for utilization of glucose significantly increased the GOS purity from 56% to ≥85%. The product consisted of tri- to penta-galactooligosaccharides. Trisaccharides were the main component of the reaction mixture. A maximum productivity of 10.9 g/L/hr was obtained under the optimum conditions.
{"title":"A novel process for the production of high-purity galactooligosaccharides (GOS) using consortium of microbes","authors":"Rengarajan Saravanan, Shajahan Shubethar, S. Narayanan, M. Jain, S. Lade, D. Jadhav, Palamalai Maheswaran, Uday K Avalakki, A. Dubey","doi":"10.1080/10826068.2016.1207082","DOIUrl":"https://doi.org/10.1080/10826068.2016.1207082","url":null,"abstract":"ABSTRACT Galactooligosaccharides (GOS) are nondigestible dietary fibers which have a beneficial effect on human health by promoting the growth of probiotic bacteria in the gut. In addition, other health benefits have been reported from oligosaccharides consumption such as stimulation of intestinal mobility, colon cancer prevention, mineral absorption as well as protection against certain pathogenic bacterial infections. The goal of this research was to develop an efficient biotransformation system using a consortium of microbes for the production of ≥85% pure GOS and reusing the cell biomass in repeated cycles of biotransformation. Production of GOS by lactose transgalactosylation using whole cells of Sporobolomyces singularis MTCC 5491 as a source of β-galactosidase and monosaccharides utilization by yeast isolate (NUTIDY007) were studied. For increasing the purity of GOS, growth and bioconversion parameters on the transgalactosylation by the whole cells were investigated. Further, continuous production of GOS was studied in a reactor with microfiltration membrane system. A maximum GOS purity of 42% was achieved using single culture of S. singularis. Under optimized conditions, single culture of S. singularis produced a maximum of 56% pure GOS. Addition of second culture to the reaction mixture for utilization of glucose significantly increased the GOS purity from 56% to ≥85%. The product consisted of tri- to penta-galactooligosaccharides. Trisaccharides were the main component of the reaction mixture. A maximum productivity of 10.9 g/L/hr was obtained under the optimum conditions.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"101 1","pages":"245 - 253"},"PeriodicalIF":0.0,"publicationDate":"2017-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79201269","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 : 2017-03-01DOI: 10.1080/10826068.2017.1286601
Géssyca P A Soares, K. T. Souza, L. Vilela, R. Schwan, D. R. Dias
ABSTRACT Flavor compounds are commonly obtained from chemical synthesis or extracted from plants. These sources have disadvantages, such as racemic mixture generation, more steps to yield the final product, low yield, and high cost, making the microbial fermentation an alternative and potential way to obtain flavor compounds. The most important lactone for flavor application is γ-decalactone, which has an aroma of peach and can be obtained by ricinoleic acid biotransformation through yeast peroxisomal β-oxidation. The aim of this work was to use crude glycerol, a residual biodiesel industry, for the production of bioaroma from two different yeasts. Yarrowia lipolytica CCMA 0357 and Lindnera saturnus CCMA 0243 were grown at different concentrations (10, 20, and 30% w/v) of substrates (castor oil and crude glycerol) for γ-decalactone production. L. saturnus CCMA 0243 produced higher concentration of y-decalactone (5.8 g/L) in crude glycerol, whereas Y. lipolytica CCMA 0357 showed a maximum production in castor oil (3.5 g/L). Crude glycerol showed better results for γ-decalactone production when compared to castor oil. L. saturnus CCMA 0243 has been shown to have a high potential for γ-decalactone production from crude glycerol.
{"title":"γ-decalactone production by Yarrowia lipolytica and Lindnera saturnus in crude glycerol","authors":"Géssyca P A Soares, K. T. Souza, L. Vilela, R. Schwan, D. R. Dias","doi":"10.1080/10826068.2017.1286601","DOIUrl":"https://doi.org/10.1080/10826068.2017.1286601","url":null,"abstract":"ABSTRACT Flavor compounds are commonly obtained from chemical synthesis or extracted from plants. These sources have disadvantages, such as racemic mixture generation, more steps to yield the final product, low yield, and high cost, making the microbial fermentation an alternative and potential way to obtain flavor compounds. The most important lactone for flavor application is γ-decalactone, which has an aroma of peach and can be obtained by ricinoleic acid biotransformation through yeast peroxisomal β-oxidation. The aim of this work was to use crude glycerol, a residual biodiesel industry, for the production of bioaroma from two different yeasts. Yarrowia lipolytica CCMA 0357 and Lindnera saturnus CCMA 0243 were grown at different concentrations (10, 20, and 30% w/v) of substrates (castor oil and crude glycerol) for γ-decalactone production. L. saturnus CCMA 0243 produced higher concentration of y-decalactone (5.8 g/L) in crude glycerol, whereas Y. lipolytica CCMA 0357 showed a maximum production in castor oil (3.5 g/L). Crude glycerol showed better results for γ-decalactone production when compared to castor oil. L. saturnus CCMA 0243 has been shown to have a high potential for γ-decalactone production from crude glycerol.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"118 1","pages":"633 - 637"},"PeriodicalIF":0.0,"publicationDate":"2017-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89434009","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 : 2017-02-28DOI: 10.1080/10826068.2017.1286599
He Ni, Xiao-Min Fan, Haojia Guo, Jianrong Liang, Qing-Rong Li, Liu Yang, Hui Li, Haihang Li
ABSTRACT The urban wastewater treatment industry produces a large amount of excess activated sludge which is mainly composed of microbial biomass and costly to be disposed. In this research, a comprehensive utilization of activated sludge was developed by sequentially extracting hydrolytic enzymes and polyhydroxyalkanoates (PHAs), and the residue was used to prepare water-retaining organic fertilizer. The sludge was extracted with fourfold H2O-containing 1% Triton X-100 with the yield of 66.7% protease activity. The enzyme solution was precipitated in 80% acetone and vacuum dried at 40°C at the dried enzyme yield of 2.4 g/kg wet sludge. The enzyme product contains collagenase, lipase, amylase, and cellulase activities, which are good compound enzymes to feed. The PHAs were extracted with 30% sodium hypoclorite:chloroform (1:3). The PHA solution was decolored and dried, and pure white PHAs were obtained at the yield of 70.1 g/kg wet sludge. The residue was used to prepare water-retaining organic fertilizer at the optimal condition. The fertilizer absorbs 131.3-fold distilled water and had good performance in water retention and can effectively slow down the loss of soil moisture when added into soil. This work provides a simple and practical approach for comprehensive utilizing activated sludge with significant economic benefits.
{"title":"Comprehensive utilization of activated sludge for the preparation of hydrolytic enzymes, polyhydroxyalkanoates, and water-retaining organic fertilizer","authors":"He Ni, Xiao-Min Fan, Haojia Guo, Jianrong Liang, Qing-Rong Li, Liu Yang, Hui Li, Haihang Li","doi":"10.1080/10826068.2017.1286599","DOIUrl":"https://doi.org/10.1080/10826068.2017.1286599","url":null,"abstract":"ABSTRACT The urban wastewater treatment industry produces a large amount of excess activated sludge which is mainly composed of microbial biomass and costly to be disposed. In this research, a comprehensive utilization of activated sludge was developed by sequentially extracting hydrolytic enzymes and polyhydroxyalkanoates (PHAs), and the residue was used to prepare water-retaining organic fertilizer. The sludge was extracted with fourfold H2O-containing 1% Triton X-100 with the yield of 66.7% protease activity. The enzyme solution was precipitated in 80% acetone and vacuum dried at 40°C at the dried enzyme yield of 2.4 g/kg wet sludge. The enzyme product contains collagenase, lipase, amylase, and cellulase activities, which are good compound enzymes to feed. The PHAs were extracted with 30% sodium hypoclorite:chloroform (1:3). The PHA solution was decolored and dried, and pure white PHAs were obtained at the yield of 70.1 g/kg wet sludge. The residue was used to prepare water-retaining organic fertilizer at the optimal condition. The fertilizer absorbs 131.3-fold distilled water and had good performance in water retention and can effectively slow down the loss of soil moisture when added into soil. This work provides a simple and practical approach for comprehensive utilizing activated sludge with significant economic benefits.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"90 1","pages":"611 - 618"},"PeriodicalIF":0.0,"publicationDate":"2017-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79405943","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 : 2017-02-13DOI: 10.1080/10826068.2016.1275014
A. Abdelhameed, S. Nusrat, S. Paliwal, M. Zaman, Nida Zaidi, R. H. Khan
ABSTRACT Drug and protein interaction provides a structural guideline in the rational drug designing and in the synthesis of new and improved drugs with greater efficacy. We have examined here the interaction tendency and mechanism of nintedanib (NTB), an anticancer drug (tyrosine kinase inhibitor) with bovine serum albumin (BSA), by spectroscopic techniques. The decline in Stern–Volmer quenching constants and binding constant with the temperature rise suggests that BSA forms a complex with NTB. Binding constant obtained by modified Stern–Volmer equation at 3 temperatures was realized to be of the order of ~104 M−1. Negative ΔG (~−5.93 kcal mol−1), ΔH (−3.74 kcal mol−1), and ΔS (−1.50 kcal mol−1) values exhibited a spontaneous and exothermic reaction between BSA and NTB. NTB molecule interacts with BSA by forming hydrogen bonds, as elucidated by fluorescence results. Moreover, a minor increment in the helical conformation of BSA upon its binding to NTB was observed by circular dichroism spectroscopy. The modification in protein’s symmetry and a decline in hydrodynamic radii were observed in the presence of NTB (from ~3.6 to ~3 nm) as obtained by the dynamic light scattering measurement results. GRAPHICAL ABSTRACT
{"title":"A multitechnique approach to probe the interaction of a therapeutic tyrosine kinase inhibitor nintedanib and bovine serum albumin","authors":"A. Abdelhameed, S. Nusrat, S. Paliwal, M. Zaman, Nida Zaidi, R. H. Khan","doi":"10.1080/10826068.2016.1275014","DOIUrl":"https://doi.org/10.1080/10826068.2016.1275014","url":null,"abstract":"ABSTRACT Drug and protein interaction provides a structural guideline in the rational drug designing and in the synthesis of new and improved drugs with greater efficacy. We have examined here the interaction tendency and mechanism of nintedanib (NTB), an anticancer drug (tyrosine kinase inhibitor) with bovine serum albumin (BSA), by spectroscopic techniques. The decline in Stern–Volmer quenching constants and binding constant with the temperature rise suggests that BSA forms a complex with NTB. Binding constant obtained by modified Stern–Volmer equation at 3 temperatures was realized to be of the order of ~104 M−1. Negative ΔG (~−5.93 kcal mol−1), ΔH (−3.74 kcal mol−1), and ΔS (−1.50 kcal mol−1) values exhibited a spontaneous and exothermic reaction between BSA and NTB. NTB molecule interacts with BSA by forming hydrogen bonds, as elucidated by fluorescence results. Moreover, a minor increment in the helical conformation of BSA upon its binding to NTB was observed by circular dichroism spectroscopy. The modification in protein’s symmetry and a decline in hydrodynamic radii were observed in the presence of NTB (from ~3.6 to ~3 nm) as obtained by the dynamic light scattering measurement results. GRAPHICAL ABSTRACT","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"62 1","pages":"655 - 663"},"PeriodicalIF":0.0,"publicationDate":"2017-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89939889","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 : 2017-02-07DOI: 10.1080/10826068.2016.1181086
D. R. Olicón-Hernández, P. Vazquez-Landaverde, R. Cruz-Camarillo, L. I. Rojas-Avelizapa
ABSTRACT Bacillus thuringiensis is a nonhuman pathogen bacterium that is used as a fungal and insect biocontrol agent. Because of its environmental interaction, it possesses several extracellular enzymes that are able to degrade chitin and chitosan, two of the most important polymers because of their application in numerous fields. However, in recent years, it has been observed that oligosaccharides from the enzymatic degradation of chitosan have important benefits for human health. Comparison and exploration of the production of chito-oligosaccharides from different sources of chitosan will improve the process parameters and expand the biotechnology based in these molecules. This study shows the production of chito-oligosaccharides from three different sources of colloidal chitosan and conducts a qualitative–quantitative comparison between them, using the extracellular enzyme of B. thuringiensis. We found that in the three substrates, it is possible to get a mixture of chito-oligosaccharides from dimer to hexamer in a concentration range from 0.72 to 8.09 mg · g−1 of original substrate. The best substrate to obtain these molecules was commercial chitosan as it has the highest production yields.
{"title":"Comparison of chito-oligosaccharide production from three different colloidal chitosans using the endochitonsanolytic system of Bacillus thuringiensis","authors":"D. R. Olicón-Hernández, P. Vazquez-Landaverde, R. Cruz-Camarillo, L. I. Rojas-Avelizapa","doi":"10.1080/10826068.2016.1181086","DOIUrl":"https://doi.org/10.1080/10826068.2016.1181086","url":null,"abstract":"ABSTRACT Bacillus thuringiensis is a nonhuman pathogen bacterium that is used as a fungal and insect biocontrol agent. Because of its environmental interaction, it possesses several extracellular enzymes that are able to degrade chitin and chitosan, two of the most important polymers because of their application in numerous fields. However, in recent years, it has been observed that oligosaccharides from the enzymatic degradation of chitosan have important benefits for human health. Comparison and exploration of the production of chito-oligosaccharides from different sources of chitosan will improve the process parameters and expand the biotechnology based in these molecules. This study shows the production of chito-oligosaccharides from three different sources of colloidal chitosan and conducts a qualitative–quantitative comparison between them, using the extracellular enzyme of B. thuringiensis. We found that in the three substrates, it is possible to get a mixture of chito-oligosaccharides from dimer to hexamer in a concentration range from 0.72 to 8.09 mg · g−1 of original substrate. The best substrate to obtain these molecules was commercial chitosan as it has the highest production yields.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"20 1","pages":"116 - 122"},"PeriodicalIF":0.0,"publicationDate":"2017-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81033125","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 : 2017-02-01DOI: 10.1080/10826068.2016.1275012
V. Krishnakumar, S. Gupta
ABSTRACT Tau is an intrinsically disordered protein responsible for maintaining the structure and stability of axonal microtubules. However, in certain disease conditions including Alzheimer’s disease, tau protein may undergo biochemical and structural changes to form intracellular aggregates. Since tau is a proline- and arginine-rich eukaryotic protein, heterologous expression in Escherichia coli often results in poor yield and has been a major technical challenge. In the current work, we have improved the expressed yield of tau by overcoming codon bias problem and established a simplified protocol for efficient extraction. The reported method has two distinct features: (i) enhanced tau expression (upto eightfold) by supplementing deficient tRNAs that aid in rapid translation and (ii) direct boiling of expressed E. coli cells to extract tau with no separate cell lysis step. We further demonstrate that tau extracted by the direct boiling method is similar to tau purified by size-exclusion chromatography exhibiting similar structural and biophysical characteristics including aggregation propensity. Since morphologies and in vitro toxicity of fibrillar tau aggregates were also similar, tau extracted by the one-step direct boiling method can be used for tau aggregation assays without any additional purification.
{"title":"Simplified method to obtain enhanced expression of tau protein from E. coli and one-step purification by direct boiling","authors":"V. Krishnakumar, S. Gupta","doi":"10.1080/10826068.2016.1275012","DOIUrl":"https://doi.org/10.1080/10826068.2016.1275012","url":null,"abstract":"ABSTRACT Tau is an intrinsically disordered protein responsible for maintaining the structure and stability of axonal microtubules. However, in certain disease conditions including Alzheimer’s disease, tau protein may undergo biochemical and structural changes to form intracellular aggregates. Since tau is a proline- and arginine-rich eukaryotic protein, heterologous expression in Escherichia coli often results in poor yield and has been a major technical challenge. In the current work, we have improved the expressed yield of tau by overcoming codon bias problem and established a simplified protocol for efficient extraction. The reported method has two distinct features: (i) enhanced tau expression (upto eightfold) by supplementing deficient tRNAs that aid in rapid translation and (ii) direct boiling of expressed E. coli cells to extract tau with no separate cell lysis step. We further demonstrate that tau extracted by the direct boiling method is similar to tau purified by size-exclusion chromatography exhibiting similar structural and biophysical characteristics including aggregation propensity. Since morphologies and in vitro toxicity of fibrillar tau aggregates were also similar, tau extracted by the one-step direct boiling method can be used for tau aggregation assays without any additional purification.","PeriodicalId":20393,"journal":{"name":"Preparative Biochemistry and Biotechnology","volume":"23 1","pages":"530 - 538"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74001658","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}
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