Applied biochemistry and biotechnology, Jan 21, 2015
Genetic modifications are considered as one of the most important technologies for improving ferm... more Genetic modifications are considered as one of the most important technologies for improving fermentative hydrogen yield. Herein, we overexpress fhlA and pncB genes from Klebsiella HQ-3 independently to enhance hydrogen molar yield. HQ-3-fhlA/pncB strain is developed by manipulation of pET28-Pkan/fhlA Kan(r) and pBBR1-MCS5/pncB Gm(r) as expression vectors to examine the synchronous effects of fhlA and pncB. Optimization of anaerobic batch fermentations is achieved and the maximum yield of biohydrogen (1.42 mol H2/mol of glucose) is produced in the range of pH 6.5-7.0 at 33-37 °C. Whole cell H2 yield is increased up to 40 % from HQ-3-fhlA/pncB, as compared with HQ-3-fhlA 20 % and HQ-3-pncB 12 % keeping HQ-3-C as a control. Mechanism of improved H2 yield is studied in combination with metabolic flux analysis by measuring glucose consumption and other metabolites including formate, succinate, 2,3 butanediol, lactate, acetate, ethanol, and hydrogen. The results suggest that under transi...
In this study, the hydrogenase-3 gene cluster (hycDEFGH) was isolated and identified from Enterob... more In this study, the hydrogenase-3 gene cluster (hycDEFGH) was isolated and identified from Enterobacter aerogenes CCTCC AB91102. All gene products were highly homologous to the reported bacterial hydrogenase-3 (Hyd-3) proteins. The genes hycE, hycF, hycG encoding the subunits of hydrogenase-3 were targeted for genetic knockout to inhibit the FHL hydrogen production pathway via the Red recombination system, generating three mutant strains AB91102-E (ΔhycE), AB91102-F (ΔhycF) and AB91102-G (ΔhycG). Deletion of the three genes affected the integrity of hydrogenase-3. The hydrogen production experiments with the mutant strains showed that no hydrogen was detected compared with the wild type (0.886mol/mol glucose), demonstrating that knocking out any of the three genes could inhibit NADH hydrogen production pathway. Meanwhile, the metabolites of the mutant strains were significantly changed in comparison with the wild type, indicating corresponding changes in metabolic flux by mutation. Additionally, the activity of NADH-mediated hydrogenase was found to be nil in the mutant strains. The chemostat experiments showed that the NADH/NAD(+) ratio of the mutant strains increased nearly 1.4-fold compared with the wild type. The NADH-mediated hydrogenase activity and NADH/NAD(+) ratio analysis both suggested that NADH pathway required the involvement of the electron transport chain of hydrogenase-3.
JPMA. The Journal of the Pakistan Medical Association, 2005
To investigate the disturbances of serum and red cell electrolytes in association with membrane N... more To investigate the disturbances of serum and red cell electrolytes in association with membrane Na(+)-K(+)- ATPase activity as well as the status of serum Urea, Creatinine and osmolality in normotensive diabetic and hypertensive diabetic patients. Thirty normotensive and thirty hypertensive patients (age and sex matched) were selected along with thirty control subjects. Erythrocytes were isolated from freshly drawn blood samples, washed and used for the estimation of sodium and potassium concentrations using flame photometer (Corning 410). Erythrocyte membranes were prepared for the estimation of Na(+)-K(+)-ATPase activity in terms of inorganic phosphate released/mg protein/hour. Serum glucose, creatinine and urea were determined by well-documented ortho toulidine, Jaffe's and diacetyl monoxime methods respectively. Osmomat 030 was used to estimate the plasma osmolality. The intra-erythrocyte sodium, serum glucose, urea, creatinine and osmolality were increased significantly in ...
This study was designed to study the relationship between serum nitric oxide and sialic acid in p... more This study was designed to study the relationship between serum nitric oxide and sialic acid in patients of diabetic nephropathy. Total 210 diabetic patients including 115 males and 95 females, suffering from diabetes and nephropathy (DN) were selected followed by informed consent and approval from institutional ethical committee. Equal number of age and sex matched normal healthy subjects were selected without any known history of hyperglycemia, hypertension and renal insufficiency as controls. Fasting blood samples from patients and controls were collected and analyzed for serum nitric oxide, sialic acid, fasting blood glucose (FBG), serum urea, creatinine, HbA1c and golmerular filtration rate (GFR). The raised levels (p<0.05) of systolic and diastolic blood pressures, BMI, FBG, HbA1c, serum urea, creatinine and sialic acid were noted in DN patients as compared to controls. Significantly lower levels of GFR and serum nitric oxide (p<0.05) were observed in DN patients as comp...
Salivary factors have been studied for their effects on the process of dental caries in patients ... more Salivary factors have been studied for their effects on the process of dental caries in patients of diabetes mellitus type 2. In this study, protective role of salivary pH, salivary flow rate, and salivary calcium is assessed in the patients of diabetes mellitus type 2 with dental caries. The samples of saliva were collected from 400 patients of diabetes mellitus type 2 and 300 age- and sex- matched controls after getting informed consent. All the subjects were classified into four groups according to age. The severity of dental caries was counted by decayed, missed, and filled teeth (DMFT) score. The salivary pH, flow rate, and calcium levels were found to be low in patients as compared to controls. The levels of fasting blood sugar, HbA1c, and DMFT score were found to be significantly high in patients than controls. The glycemic factors were significantly correlated with salivary factors indicating their influence on progression of caries in diabetes. On the basis of findings, it ...
Applied biochemistry and biotechnology, Jan 21, 2015
Genetic modifications are considered as one of the most important technologies for improving ferm... more Genetic modifications are considered as one of the most important technologies for improving fermentative hydrogen yield. Herein, we overexpress fhlA and pncB genes from Klebsiella HQ-3 independently to enhance hydrogen molar yield. HQ-3-fhlA/pncB strain is developed by manipulation of pET28-Pkan/fhlA Kan(r) and pBBR1-MCS5/pncB Gm(r) as expression vectors to examine the synchronous effects of fhlA and pncB. Optimization of anaerobic batch fermentations is achieved and the maximum yield of biohydrogen (1.42 mol H2/mol of glucose) is produced in the range of pH 6.5-7.0 at 33-37 °C. Whole cell H2 yield is increased up to 40 % from HQ-3-fhlA/pncB, as compared with HQ-3-fhlA 20 % and HQ-3-pncB 12 % keeping HQ-3-C as a control. Mechanism of improved H2 yield is studied in combination with metabolic flux analysis by measuring glucose consumption and other metabolites including formate, succinate, 2,3 butanediol, lactate, acetate, ethanol, and hydrogen. The results suggest that under transi...
In this study, the hydrogenase-3 gene cluster (hycDEFGH) was isolated and identified from Enterob... more In this study, the hydrogenase-3 gene cluster (hycDEFGH) was isolated and identified from Enterobacter aerogenes CCTCC AB91102. All gene products were highly homologous to the reported bacterial hydrogenase-3 (Hyd-3) proteins. The genes hycE, hycF, hycG encoding the subunits of hydrogenase-3 were targeted for genetic knockout to inhibit the FHL hydrogen production pathway via the Red recombination system, generating three mutant strains AB91102-E (ΔhycE), AB91102-F (ΔhycF) and AB91102-G (ΔhycG). Deletion of the three genes affected the integrity of hydrogenase-3. The hydrogen production experiments with the mutant strains showed that no hydrogen was detected compared with the wild type (0.886mol/mol glucose), demonstrating that knocking out any of the three genes could inhibit NADH hydrogen production pathway. Meanwhile, the metabolites of the mutant strains were significantly changed in comparison with the wild type, indicating corresponding changes in metabolic flux by mutation. Additionally, the activity of NADH-mediated hydrogenase was found to be nil in the mutant strains. The chemostat experiments showed that the NADH/NAD(+) ratio of the mutant strains increased nearly 1.4-fold compared with the wild type. The NADH-mediated hydrogenase activity and NADH/NAD(+) ratio analysis both suggested that NADH pathway required the involvement of the electron transport chain of hydrogenase-3.
JPMA. The Journal of the Pakistan Medical Association, 2005
To investigate the disturbances of serum and red cell electrolytes in association with membrane N... more To investigate the disturbances of serum and red cell electrolytes in association with membrane Na(+)-K(+)- ATPase activity as well as the status of serum Urea, Creatinine and osmolality in normotensive diabetic and hypertensive diabetic patients. Thirty normotensive and thirty hypertensive patients (age and sex matched) were selected along with thirty control subjects. Erythrocytes were isolated from freshly drawn blood samples, washed and used for the estimation of sodium and potassium concentrations using flame photometer (Corning 410). Erythrocyte membranes were prepared for the estimation of Na(+)-K(+)-ATPase activity in terms of inorganic phosphate released/mg protein/hour. Serum glucose, creatinine and urea were determined by well-documented ortho toulidine, Jaffe's and diacetyl monoxime methods respectively. Osmomat 030 was used to estimate the plasma osmolality. The intra-erythrocyte sodium, serum glucose, urea, creatinine and osmolality were increased significantly in ...
This study was designed to study the relationship between serum nitric oxide and sialic acid in p... more This study was designed to study the relationship between serum nitric oxide and sialic acid in patients of diabetic nephropathy. Total 210 diabetic patients including 115 males and 95 females, suffering from diabetes and nephropathy (DN) were selected followed by informed consent and approval from institutional ethical committee. Equal number of age and sex matched normal healthy subjects were selected without any known history of hyperglycemia, hypertension and renal insufficiency as controls. Fasting blood samples from patients and controls were collected and analyzed for serum nitric oxide, sialic acid, fasting blood glucose (FBG), serum urea, creatinine, HbA1c and golmerular filtration rate (GFR). The raised levels (p<0.05) of systolic and diastolic blood pressures, BMI, FBG, HbA1c, serum urea, creatinine and sialic acid were noted in DN patients as compared to controls. Significantly lower levels of GFR and serum nitric oxide (p<0.05) were observed in DN patients as comp...
Salivary factors have been studied for their effects on the process of dental caries in patients ... more Salivary factors have been studied for their effects on the process of dental caries in patients of diabetes mellitus type 2. In this study, protective role of salivary pH, salivary flow rate, and salivary calcium is assessed in the patients of diabetes mellitus type 2 with dental caries. The samples of saliva were collected from 400 patients of diabetes mellitus type 2 and 300 age- and sex- matched controls after getting informed consent. All the subjects were classified into four groups according to age. The severity of dental caries was counted by decayed, missed, and filled teeth (DMFT) score. The salivary pH, flow rate, and calcium levels were found to be low in patients as compared to controls. The levels of fasting blood sugar, HbA1c, and DMFT score were found to be significantly high in patients than controls. The glycemic factors were significantly correlated with salivary factors indicating their influence on progression of caries in diabetes. On the basis of findings, it ...
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Papers by Muhammad Faisal Jawed