Career Episode 1
1.0 This career episode talks about my major final year project during my bachelor’s degree.
The details of the project are given below:
Project Title: Polymerisation of Phenol using Radish Peroxidase.
Duration: 6 months (January 2013 to June 2013)
Geographical Location: Bellary – 583104, Karnataka, India.
Name of Organisation: Bellary Institute of Technology and Management.
Position Title: Project Associate
1.1 The project “POLYMERISATION OF PHENOL USING RADISH PEROXIDASE” was a Major project undertaken as a part of my course during the final year of my graduate degree. Three students including myself from the Department of Biotechnology Engineering worked on this project under the guidance of a Project guide from the department.
The prime objective of this Project was to remove phenols and phenolic compounds from waste water streams generated by many industries such as coal conversion, resins, plastic, petroleum refining, textiles, dyes The project reporting and timeline strictly followed the specification rules and regulations rolled out by the department and the university. Final evaluation of the project was done by the head of the department, Project guide and University External. The final evaluation included a demonstration of complete project, power point presentation with viva and report submission. The project was also accepted and published in National Conference on Recent Advances in Civil Engineering (Wastewater Treatment technologies section).
1.2 Personal Workspace
Statements of Duties
My duties during the course of the project involved:
– Assisting in research to understand polymerisation of Phenol using enzymatic process.
– Studying current methods and anomalies and devise a new method to overcome the problems in existing techniques.
– Gather data for analysis and to implement in testing
– Analysis of the enzyme using Bioinformatics tools.
– Developing a Project plan.
– Assisting in development of project Report preparation, Power point preparation.
– Providing detailed explanation on polymerisation of Phenol with graph representation.
C Personal Engineering Activity
1.3 Reporting Structure of the Project
1.4 Problem found
Phenol is an organic pollutant present in wastewater from various industries such as petroleum refining, resin and plastic, leather and textile manufacturing, chemical and petrochemical plants etc. The presence of phenols in the environment poses a significant risk to aquatic biota. This makes it a major focus of environment and health pollution control. Hence, this research to removal of phenol from aqueous solution addresses an important problem.
1.5 Studying current methods and anomalies and devise a new method to overcome the problems in existing techniques
Phenols can be removed from industrial effluents by physiochemical methods such as ozonation, Fenton’s agent, UV or hydrogen peroxide, but these treatments are usually complex and expensive. For these reasons, we have opted for enzymatic approach which will target aromatic pollutants as a potential and viable alternative to conventional methods due to their highly sensitive nature. On research we found that Peroxidase is that enzyme which have wide spectrum applications in a number of industrial process and transforms toxic compounds of industrial origin such as aromatic compound (Phenol) to preserver the quality of water. This process is defined as Polymerisation of Phenol using H2O2 in the presence of Enzyme which will breakdown the phenol and remove about 98% from water.
1.6 Implementation of data in our testing.
I extracted Radish Peroxidase enzyme from fresh Radish obtained from market and further centrifuged at 6000 rpm at 25°C for 30 mins. Further the activity of RP enzyme was tested according to the method of Worthington. Phenol containing water was treated with RP enzyme at different substrate concentration, temperature, pH, PEG, Inhibitors and monitored the effect of these on OD meter at 30mins time interval. I prepared graph for measurement obtained during the test. The results are demonstrated in the section 1.8.
1.7 Developing Project Plan
During the duration of the project, the regular classes were still going on and we were running out of time. I was asked to develop a project plan which would include the work division among the team members. I took the responsibility of research, preparing reports as well as power point presentation. I suggested the use of Gantt chart would be the best option for developing project plan because it’s easy to understand and enable better project control by promoting clearer communication.
Approval of Proposal
Collection of Data from resources
Materials and Requirements
Preparation of Report and PPT
This is the Gantt chart prepared by me and circulated among the team members. This has helped us in dividing each activity between the team mates. I researched a lot for selecting a Specific project where we could apply our subject knowledge, cost effective and match our time constraints. Availability of materials required for the project was another challenge we had to go through.
1.8 Phenol Removal explanation with Graphical representation.
Removal of phenol using Radish peroxidase enzyme is been studied using various factors such as Effect of Substrate concentration, temperature, pH, PEG concentration and inhibitor. The extracted Radish Peroxidase enzyme polymerized phenol to a maximum of 98% for the concentration of 100 mg/L. The time duration taken up by the RP was about 30 min and the results obtained for favourable polymerization were at conditions 30 °C, pH – 6.8, Activator (PEG) concentration of 40 mg/L and inhibitor (EDTA) of 10 mM concentration.
Graph 1: Effect of Substrate Concentration
The initial concentration of Phenol increased from 50 to 1200 mg/L, the amount of phenol transformation catalysed by 20U immobilized HRP was 26 mg at most.
Graph 2: Effect of Temperature
Maximum removal of phenol was observed at 40 °C.
Graph 3: Effect of pH
The maximum phenol removal was observer in the buffer of pH 6.8
Graph 4: Effect pf PEG
Table 1: Effect of Inhibitors
The activity of enzyme in polymerizing the phenol present in wastewater was determined in presence of different inhibitors like Salicylic acid, Sodium Chloride and EDTA with 10 nM concentration. The results obtained are shown in the Table 1.
1.9 Analysis of RP enzyme using Bioinformatics tools.
It was important to predict the function of Radish peroxidase enzyme and for that I have carried out Insilco study of Radish Peroxidase Enzyme using Bioinformatics Tools such as ProtParam, SOPMA, ESyPred3D, Rasmol, and ProFunc.
Fig: 3D Structure of Radish Peroxidase
Fig: Active sites of Radish Peroxidase.
1.10 Assisting in development of Final Project Report preparation and Power point preparation
This task was assigned to me as I have a very good hold on detail, document preparation and Power point preparation. I was keener on researching as I had to and made sure that we opted for cost effective project which meet our timelines and have all raw materials readily available for the experiments. I also used my Genomics and Bioinformatics skills to carry the Insilco study of Radish Peroxidase enzyme.
I made sure that every resource I researched was from reliable source and mentioned the same in the Final Report under References. In guidance with my Project guide I came out with final Project report which included the each and every process involved during the project and submitted to our Head of the Department.
I sorted out the complete project into three parts and helped them in getting ready for power point presentation by assigning them their task to explain as each one of us were evaluated on out presentation skills as well. The success of the project was realised when our project was accepted and published in National Conference on Recent Advances in Civil Engineering (Wastewater Treatment Technologies section). Since I was solely involved in research, I have taken the responsibility of explaining the deep analysis of the study in a viva session. The presentation went very well.
1.10 This project gave a real-time experience of how to analyse things and helped me using my subject knowledge, finding the connections between them and making a detailed report out of it. Though there were a lot of time constraints as we had our classes and exams running alongside, but it gave a satisfactory work experience. My deep research on Insilco Study of RP enzyme has improved my Bioinformatics skills and Biochemistry skills. The project plan developed my me have not only given a clear picture of our tasks but also motivated us to deliver our project in time confidently.
Technical Paper Accepted for Publication
1.11 Our project entitled “Polymerization of Phenol Using Radish Peroxidase” has been accepted and published in National Conference on Recent Advances in Civil Engineering (Wastewater treatment technologies section) and “The International Institute for Science, Technology and Education (IISTE)”. The aim of the institute is Accelerating Global Knowledge Sharing.