14BBI30 - Biotechnology lab 1

Course specification
Course title		Biotechnology lab 1
Acronym		14BBI30
Study programme		Biochemical Engineering and Biotechnology
Module
Lecturer (for classes)		prof. dr Mirjana Rajilić-Stojanović
Lecturer/Associate (for practice)		prof. dr Mirjana Rajilić-Stojanović doc. dr Sonja Jakovetić Tanasković
Lecturer/Associate (for OTC)
ESPB		3.0	Status
Condition			Облик условљености
The goal		The main goal is to enable students to use biochemical methods for identification and quantitative determination the most important natural compounds, which are prerequisite for following the course of biochemical processes. Methods for determination of protein concentration, protein purification, and enzyme activity assays will be learned and subsequently applied in microbial enzyme production and determination of kinetic constants.
The outcome		Students will become skilled at techniques and methods used in biochemical laboratory. During the course students gain basic knowledge on extraction of bioactive cell compounds (vitamins and proteins) and their further purification, determination and characterization. Moreover, students will be learned to process, analyze and present experimental data.
Contents
Contents of lectures		Color reactions in qualitative protein analysis (general and amino acid specific). Protein precipitation. Quantitative protein analysis (spectrophotometry and fluorometry assays). Determination of reducing sugars and total sugars concentration. Vitamin C volumetric assays. Cell disruption methods. Protein separation and purification. Electrophoresis in protein detection and characterization. Calculation of enzyme activity and determination of kinetic constants.
Contents of exercises		Quantification of vitamin C using volumetric method. Determination of reducing sugars. Protein identification усинг color reactions and quantitative determination with Lowry and/or Bradford method. Extraction and purification of intracellular baker’s yeast invertase. Spectroscopic enzyme activity assay. Calculation of purification factor. Electrophoretic characterization of proteins. Study of the effect of experimental factors (pH, temperature, initital substrate concentration, enzyme concentration) on enzyme activity. Kinetic constant calculation and graphical presentation of results using MS Excell software.
Literature
Mikkelsen S. R., Corton E: Bioanalytical chemistry, Wiley-Interscience, 2004. (Original title) Šiler-Marinković S., Knežević Z., Bezbradica D., Biohemijski praktikum'' (interni materijal). (Original title)
Number of hours per week during the semester/trimester/year
Lectures	Exercises	OTC	Study and Research	Other classes
1	1	2
Methods of teaching		Theoretical lectures. Laboratory practice. Problem solving and calculus.
Knowledge score (maximum points 100)
Pre obligations		Points	Final exam	Points
Activites during lectures			Test paper	40
Practical lessons		20	Oral examination
Projects
Colloquia		40
Seminars