Search results for: biotechnology

Authors: Wenjun Zhang, Yunqing Du, Ming L. Wang

Abstract:

Noninvasive diagnostics of diseases via breath analysis has attracted considerable scientific and clinical interest for many years and become more and more promising with the rapid advancements in nanotechnology and biotechnology. The volatile organic compounds (VOCs) in exhaled breath, which are mainly blood borne, particularly provide highly valuable information about individuals’ physiological and pathophysiological conditions. Additionally, breath analysis is noninvasive, real-time, painless, and agreeable to patients. We have developed a wireless sensor array based on single-stranded DNA (ssDNA)-functionalized single-walled carbon nanotubes (SWNT) for the detection of a number of physiological indicators in breath. Seven DNA sequences were used to functionalize SWNT sensors to detect trace amount of methanol, benzene, dimethyl sulfide, hydrogen sulfide, acetone, and ethanol, which are indicators of heavy smoking, excessive drinking, and diseases such as lung cancer, breast cancer, and diabetes. Our test results indicated that DNA functionalized SWNT sensors exhibit great selectivity, sensitivity, and repeatability; and different molecules can be distinguished through pattern recognition enabled by this sensor array. Furthermore, the experimental sensing results are consistent with the Molecular Dynamics simulated ssDNAmolecular target interaction rankings. Thus, the DNA-SWNT sensor array has great potential to be applied in chemical or biomolecular detection for the noninvasive diagnostics of diseases and personal health monitoring.

Keywords: Breath analysis, DNA-SWNT sensor array, diagnosis, noninvasive.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2785

Authors: Mun Yee Chan, Sin Ming Goh, Lisa Gaik Ai Ong

Abstract:

Approximately 10,000 different types of dyes and pigments are being used in various industrial applications yearly, which include the textile and printing industries. However, these dyes are difficult to degrade naturally once they enter the aquatic system. Their high persistency in natural environment poses a potential health hazard to all form of life. Hence, there is a need for alternative dye removal strategy in the environment via bioremediation. In this study, fungi laccase is investigated via commercial agar dyes plates and submerged fermentation to explore the application of fungi laccase in textile dye wastewater treatment. Two locally isolated basidiomycetes were screened for laccase activity using media added with commercial dyes such as 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), guaiacol and Remazol Brillant Blue R (RBBR). Isolate TBB3 (1.70±0.06) and EL2 (1.78±0.08) gave the highest results for ABTS plates with the appearance of greenish halo on around the isolates. Submerged fermentation performed on Isolate TBB3 with the productivity 3.9067 U/ml/day, whereas the laccase activity for Isolate EL2 was much lower (0.2097 U/ml/day). As isolate TBB3 showed higher laccase production, it was subjected to molecular characterization by DNA isolation, PCR amplification and sequencing of ITS region of nuclear ribosomal DNA. After being compared with other sequences in National Center for Biotechnology Information (NCBI database), isolate TBB3 is probably from species Trametes hirsutei. Further research work can be performed on this isolate by upscale the production of laccase in order to meet the demands of the requirement for higher enzyme titer for the bioremediation of textile dyes.

Keywords: Bioremediation, dyes, fermentation, laccase.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2128

Authors: Ahmad Ali Shahid, Muhammad Shakil Shaukat, Kamran Shehzad Bajwa, Abdul Qayyum Rao, Tayyab Husnain

Abstract:

Agriculture is the backbone of economy of Pakistan and cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat severe problems of insect and weed, combination of three genes namely Cry1Ac, Cry2A and EPSPS genes was transferred in locally cultivated cotton variety MNH-786 with the use of Agrobacterium mediated genetic transformation. The present study focused on the molecular screening of transgenic cotton plants at T3 generation in order to confirm integration and expression of all three genes (Cry1Ac, Cry2A and EPSP synthase) into the cotton genome. Initially, glyphosate spray assay was used for screening of transgenic cotton plants containing EPSP synthase gene at T3 generation. Transgenic cotton plants which were healthy and showed no damage on leaves were selected after 07 days of spray. For molecular analysis of transgenic cotton plants in the laboratory, the genomic DNA of these transgenic cotton plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty (Cry1Ac gene), ten out of twenty (Cry2A gene) and all twenty (EPSP synthase gene) were produced positive amplification. On the base of PCR amplification, ten transgenic plant samples were subjected to protein expression analysis through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the mRNA expression levels of Cry1Ac and EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes at T3 generation.

Keywords: Agriculture, Cotton, Transformation, Cry Genes, ELISA and PCR.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 3072

Authors: Vongsathorn Ngampuak, Yutachai Chookaew, Wipawee Dejtisakdi

Abstract:

Dunaliella salina has great potential as a system for generating commercially valuable products, including beta-carotene, pharmaceuticals, and biofuels. Our goal is to improve this potential by enhancing growth rate and other properties of D. salina under optimal growth conditions. We used ethyl methane sulfonate (EMS) to generate random mutants in D. salina KU11, a strain classified in Thailand. In a preliminary experiment, we first treated D. salina cells with 0%, 0.8%, 1.0%, 1.2%, 1.44% and 1.66% EMS to generate a killing curve. After that, we randomly picked 30 candidates from approximately 300 isolated survivor colonies from the 1.44% EMS treatment (which permitted 30% survival) as an initial test of the mutant screen. Among the 30 survivor lines, we found that 2 strains (mutant #17 and #24) had significantly improved growth rates and cell number accumulation at stationary phase approximately up to 1.8 and 1.45 fold, respectively, 2 strains (mutant #6 and #23) had significantly decreased growth rates and cell number accumulation at stationary phase approximately down to 1.4 and 1.35 fold, respectively, while 26 of 30 lines had similar growth rates compared with the wild type control. We also analyzed cell size for each strain and found there was no significant difference comparing all mutants with the wild type. In addition, mutant #24 had shown an increase of biomass accumulation approximately 1.65 fold compared with the wild type strain on day 5 that was entering early stationary phase. From these preliminary results, it could be feasible to identify D. salina mutants with significant improved growth rate, cell accumulation and biomass production compared to the wild type for the further study; this makes it possible to improve this microorganism as a platform for biotechnology application.

Keywords: Dunaliella salina, mutant, ethyl methane sulfonate, growth rate, biomass.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1803

Authors: Mário Silva, Filipa Gomes, Filipa Oliveira, Simone Morais, Cristina Delerue-Matos

Abstract:

Brown seaweeds are abundant in Portuguese coastline and represent an almost unexploited marine economic resource. One of the most common species, easily available for harvesting in the northwest coast, is Saccorhiza polyschides grows in the lowest shore and costal rocky reefs. It is almost exclusively used by local farmers as natural fertilizer, but contains a substantial amount of valuable compounds, particularly alginates, natural biopolymers of high interest for many industrial applications. Alginates are natural polysaccharides present in cell walls of brown seaweed, highly biocompatible, with particular properties that make them of high interest for the food, biotechnology, cosmetics and pharmaceutical industries. Conventional extraction processes are based on thermal treatment. They are lengthy and consume high amounts of energy and solvents. In recent years, microwave-assisted extraction (MAE) has shown enormous potential to overcome major drawbacks that outcome from conventional plant material extraction (thermal and/or solvent based) techniques, being also successfully applied to the extraction of agar, fucoidans and alginates. In the present study, acid pretreatment of brown seaweed Saccorhiza polyschides for subsequent microwave-assisted extraction (MAE) of alginate was optimized. Seaweeds were collected in Northwest Portuguese coastal waters of the Atlantic Ocean between May and August, 2014. Experimental design was used to assess the effect of temperature and acid pretreatment time in alginate extraction. Response surface methodology allowed the determination of the optimum MAE conditions: 40 mL of HCl 0.1 M per g of dried seaweed with constant stirring at 20ºC during 14h. Optimal acid pretreatment conditions have enhanced significantly MAE of alginates from Saccorhiza polyschides, thus contributing for the development of a viable, more environmental friendly alternative to conventional processes.

Keywords: Acid pretreatment, Alginate, Brown seaweed, Microwave-assisted extraction, Response surface methodology.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 3266

Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

Abstract:

One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1511

Authors: Philip Semaha, Zhongfang Lei, Ziwen Zhao, Sen Liu, Zhenya Zhang, Kazuya Shimizu

Abstract:

The increasing generation of saline wastewater through various industrial activities is becoming a global concern for activated sludge (AS) based biological treatment which is widely applied in wastewater treatment plants (WWTPs). As for the AS process, an increase in wastewater salinity has negative impact on its overall performance. The advent of conventional aerobic granular sludge (AGS) or bacterial AGS biotechnology has gained much attention because of its superior performance. The development of algal-bacterial AGS could enhance better nutrients removal, potentially reduce aeration cost through symbiotic algae-bacterial activity, and thus, can also reduce overall treatment cost. Nonetheless, the potential of salt stress to decrease biomass growth, microbial activity and nutrient removal exist. Up to the present, little information is available on saline wastewater treatment by algal-bacterial AGS. To the authors’ best knowledge, a comparison of the two AGS systems has not been done to evaluate nutrients removal capacity in the context of salinity increase. This study sought to figure out the impact of salinity on the algal-bacterial AGS system in comparison to bacterial AGS one, contributing to the application of AGS technology in the real world of saline wastewater treatment. In this study, the salt concentrations tested were 0 g/L, 1 g/L, 5 g/L, 10 g/L and 15 g/L of NaCl with 24-hr artificial illuminance of approximately 97.2 µmol m¯²s¯¹, and mature bacterial and algal-bacterial AGS were used for the operation of two identical sequencing batch reactors (SBRs) with a working volume of 0.9 L each, respectively. The results showed that salinity increase caused no apparent change in the color of bacterial AGS; while for algal-bacterial AGS, its color was progressively changed from green to dark green. A consequent increase in granule diameter and fluffiness was observed in the bacterial AGS reactor with the increase of salinity in comparison to a decrease in algal-bacterial AGS diameter. However, nitrite accumulation peaked from 1.0 mg/L and 0.4 mg/L at 1 g/L NaCl in the bacterial and algal-bacterial AGS systems, respectively to 9.8 mg/L in both systems when NaCl concentration varied from 5 g/L to 15 g/L. Almost no ammonia nitrogen was detected in the effluent except at 10 g/L NaCl concentration, where it averaged 4.2 mg/L and 2.4 mg/L, respectively, in the bacterial and algal-bacterial AGS systems. Nutrients removal in the algal-bacterial system was relatively higher than the bacterial AGS in terms of nitrogen and phosphorus removals. Nonetheless, the nutrient removal rate was almost 50% or lower. Results show that algal-bacterial AGS is more adaptable to salinity increase and could be more suitable for saline wastewater treatment. Optimization of operation conditions for algal-bacterial AGS system would be important to ensure its stably high efficiency in practice.

Keywords: Algal-bacterial aerobic granular sludge, bacterial aerobic granular sludge, nutrients removal, saline wastewater, sequencing batch reactor.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1072