Md. Azizul Haque

Cloning and expression of ophB gene encoding organophosphorus hydrolase from endophytic Pseudomonas sp. BF1-3 degrades organophosphorus pesticide chlorpyrifos.

Chlorpyrifos is an organophosphate pesticide that has adverse effect on animals and plants. We isolated endophytic bacterial strain, Pseudomonas sp. BF1-3, from balloon flower root which can hydrolyze chlorpyrifos. A gene (ophB) encoding a protein involved in chlorpyrifos degradation from this strain was cloned into Escherichia coli DH5α for confirming enzyme activity. After sequencing, total 1024bp nucleotide sequences were found in the open reading frame of ophB. The chlorpyrifos degradation patterns by E. coli DH5α (ophB) were observed. During incubation in minimal salt (M9) medium supplemented with chlorpyrifos (100mgL(-1)), the E. coli DH5α harboring ophB degraded about 97% initial chlorpyrifos (100mgL(-1)) and accumulated 86mgL(-1) 3,5,6-trichloro-2-pyridinol (TCP) within 9 days. In addition, optical density (OD) of E. coli DH5α (ophB) culture at 600nm was increased from 0.172 to 1.118 within 2 days of inoculation in the chlorpyrifos supplemented M9 medium. The estimated molecular weight of purified OphB protein was determined to be 31.4kDa by SDS-PAGE. The OphB enzyme was most active at pH 8 and an optimal temperature around 35°C. These results indicate that endophytic bacteria are supposed to be useful for biological control of environments contaminated with pesticides.

Effect of mild alkali pretreatment on structural changes of reed (Phragmites communis Trinius) straw

The effect of dilute sodium hydroxide (NaOH) on reed straw structural change at 105°C temperature was evaluated in this study. Various concentrations of NaOH (1% to 2.5%) were used for pretreatment of reed straw at 105°C for 10 min. Scanning electron microscopy, atomic force microscopy and Fourier transform infrared spectroscopy studies showed that 2% and 2.5% NaOH pretreated sample exposed more cellulose fibers compared with other treatments. The cellulose crystalline index was increased by the 1% to 2.0% NaOH treatments and slightly lowered by the 2.5% NaOH treatment due to destructing cellulose fibres. Two per cent NaOH pretreatment caused 69.9% lignin removal, whereas 2.5% NaOH pretreatment removed 72.4% lignin. Besides, reed straw, when pretreated at 2% and 2.5% NaOH, resulted 56.4% and 60.5% hemicellulose removal, respectively. However, the difference in removal of lignin and hemicellulose between 2% and 2.5% NaOH treated reed straw was very marginal. In addition, very negligible increase of cellulose level was estimated, amounting 78.8% and 76.6% in 2.5% and 2% NaOH-treated sample, respectively. Moreover, after 72 h, reducing sugar yield was 81.2% and 83.3% using enzyme loading of 15 FPU (g dry biomass)−1 and 30 IU (g dry biomass)−1 and xylanase 4 FXU (g dry biomass)−1 from 2% and 2.5% NaOH pretreated reed straw, respectively. Reducing sugar yield was increased very marginally when NaOH concentration increased from 2% to 2.5% for reed straw pretreatment. Therefore, 2% NaOH is supposed to be effective for reed straw pretreatment at this mentioned condition.

Alkali Pretreatment of Wheat Straw (Triticum aestivum) at Boiling Temperature for Producing a Bioethanol Precursor

We evaluated the effect of dilute sodium hydroxide (NaOH) on wheat straw at boiling temperature for removing lignin and increasing the yield of reducing sugar. Various concentrations of NaOH (0.5% to 2%) were used for pretreating wheat straw at 105 °C for 10 min. Scanning electron microscopy, atomic force microscopy, and Fourier transform infrared spectroscopy studies revealed that the 2% NaOH-pretreated sample exposed more cellulose fiber. The maximum respective removal of lignin and hemicellulose was 70.3% and 68.2% from the 2% NaOH-pretreated liquor. The reducing sugar yield was 84.6% using an enzyme dose containing 20 FPU of cellulase, 40 IU of β-glucosidase and 4 FXU of xylanase/g of substrate. However, 2% NaOH-treated wheat straw had the lowest crystalline index of 52.5%, due to destructured cellulose fibers. The results indicate the effectiveness of producing the bioethanol precursor from wheat straw by using 2% NaOH at boiling temperature.