Anna MARABOTTI | APPLIED BIOCHEMISTRY

cod. 0522300040

APPLIED BIOCHEMISTRY

	0522300040
	DEPARTMENT OF CHEMISTRY AND BIOLOGY "ADOLFO ZAMBELLI"
	EQF7
	CHEMISTRY
	2023/2024

CURRICULUM BIOMOLECULAR CHEMISTRY

	OBBLIGATORIO
	YEAR OF COURSE 1
	YEAR OF DIDACTIC SYSTEM 2016
	SPRING SEMESTER

TEACHING UNITS

SSD	CFU	HOURS	ACTIVITY	TYPE OF ACTIVITY
BIO/10	5	40	LESSONS	COMPULSORY SUBJECTS, CHARACTERISTIC OF THE CLASS
BIO/10	1	12	LAB	COMPULSORY SUBJECTS, CHARACTERISTIC OF THE CLASS

PROFESSORS

	ANNA MARABOTTI T Curriculum
	GAETANA PAOLELLA Curriculum

	Objectives
	THE COURSE AIMS TO PROVIDE THE KNOWLEDGE OF THE MAIN METHODS FOR THE STUDY OF THE MACROMOLECULES OF BIOLOGICAL INTEREST (PROTEINS, NUCLEIC ACIDS), WITH PARTICULAR REFERENCE TO PRODUCTION STRATEGIES, PURIFICATION AND CHARACTERIZATION OF THESE MACROMOLECULES. THE MAIN KNOWLEDGE ACQUIRED BY THE STUDENT WILL CONCERN: -THE CHEMICO-PHYSICAL FEATURES AND THE BIOLOGICAL ROLE OF BIOLOGICAL MACROMOLECULES AND HOW THESE FEATURES CAN BE USED FOR THEIR PURIFICATION AND CHARACTERIZATION; - THE DIFFERENT METHODS OF PURIFICATION OF PROTEINS AND NUCLEIC ACIDS: METHODS OF HOMOGENIZATION OF CELLS AND TISSUES, DIFFERENTIAL PRECIPITATION, CENTRIFUGATION AND DIALYSIS, CHROMATOGRAPHY; -THE DIFFERENT ANALYTICAL METHODS RELATED TO PROTEINS AND NUCLEIC ACIDS: ELECTROPHORESIS, SPECTROSCOPIC METHODS, SEQUENCING METHODS , IMMUNOHCEMICAL METHODS, TECNIQUES OF BLOTTING; - COMPUTATIONAL TECHNIQUES APPLIED TO MOLECULAR DESIGN AND THE STUDY OF MOLECULAR INTERACTIONS; -THE MAIN METHODS OF THE RECOMBINAT DNA. THE MAIN SKILLS DEVELOPED BY THE STUDENT WILL BE: -TO KNOW AND EXPOSE THE CHARACTERISTICS OF THE VARIOUS TECHNIQUES STUDIED -TO CRITICALLY EVALUATE PROTEIN PRODUCTION AND PURIFICATION PROTOCOLS; -TO EVALUATE THE STRUCTURAL AND FUNCTIONAL CHARACTERISTICS OF PROTEINS AND NUCLEIC ACIDS ON THE BASIS OF RESULTS OBTAINED WITH DIFFERENT TECHNIQUES; -TO PERFORM SIMPLE PROTOCOLS FOR PURIFICATION AND CHARACTERIZATION OF PROTEINS AND NUCLEIC ACIDS.

THE COURSE AIMS TO PROVIDE THE KNOWLEDGE OF THE MAIN METHODS FOR THE STUDY OF THE MACROMOLECULES OF BIOLOGICAL INTEREST (PROTEINS, NUCLEIC ACIDS), WITH PARTICULAR REFERENCE TO PRODUCTION STRATEGIES, PURIFICATION AND CHARACTERIZATION OF THESE MACROMOLECULES.

THE MAIN KNOWLEDGE ACQUIRED BY THE STUDENT WILL CONCERN:
-THE CHEMICO-PHYSICAL FEATURES AND THE BIOLOGICAL ROLE OF BIOLOGICAL MACROMOLECULES AND HOW THESE FEATURES CAN BE USED FOR THEIR PURIFICATION AND CHARACTERIZATION;
- THE DIFFERENT METHODS OF PURIFICATION OF PROTEINS AND NUCLEIC ACIDS: METHODS OF HOMOGENIZATION OF CELLS AND TISSUES, DIFFERENTIAL PRECIPITATION, CENTRIFUGATION AND DIALYSIS, CHROMATOGRAPHY;
-THE DIFFERENT ANALYTICAL METHODS RELATED TO PROTEINS AND NUCLEIC ACIDS: ELECTROPHORESIS, SPECTROSCOPIC METHODS, SEQUENCING METHODS , IMMUNOHCEMICAL METHODS, TECNIQUES OF BLOTTING;
- COMPUTATIONAL TECHNIQUES APPLIED TO MOLECULAR DESIGN AND THE STUDY OF MOLECULAR INTERACTIONS;
-THE MAIN METHODS OF THE RECOMBINAT DNA.

THE MAIN SKILLS DEVELOPED BY THE STUDENT WILL BE:
-TO KNOW AND EXPOSE THE CHARACTERISTICS OF THE VARIOUS TECHNIQUES STUDIED
-TO CRITICALLY EVALUATE PROTEIN PRODUCTION AND PURIFICATION PROTOCOLS;
-TO EVALUATE THE STRUCTURAL AND FUNCTIONAL CHARACTERISTICS OF PROTEINS AND NUCLEIC ACIDS ON THE BASIS OF RESULTS OBTAINED WITH DIFFERENT TECHNIQUES;
-TO PERFORM SIMPLE PROTOCOLS FOR PURIFICATION AND CHARACTERIZATION OF PROTEINS AND NUCLEIC ACIDS.

	Prerequisites
	THE STUDENT MUST HAVE ACQUIRED THE BASIC KNOWLEDGE IN BIOCHEMISTRY PROVIDED BY THE TRIENNIAL STUDY PROGRAM, WITH PARTICULAR FOCUS ON: - CHEMICAL FEATURES OF BIOLOGICAL MACROMOLECULES (PROTEINS, NUCLEIC ACIDS, LIPIDS, GLUCIDS) - ROLE OF THESE MACROMOLECULES IN THE HUMAN BODY

	Contents
	THE TEACHING INCLUDES 40 HOURS OF DIDACTICS IN THE FORM OF FRONTAL LECTURES AND GUIDED CLASSROOM EXERCISES, AND 12 HOURS OF BIOCHEMISTRY LABORATORY (1 ECTS: BY PROF. GAETANA PAOLELLA), DURING WHICH STUDENTS DIRECTLY TEST WHAT THE LECTURER INTRODUCES IN THE LECTURE (USING A LEARNING-BY-DOING APPROACH). TOPICS COVERED IN THE CLASSROOM LECTURES ARE: - STRUCTURAL FEATURES OF PROTEINS AND NUCLEIC ACIDS (APPROXIMATELY 2 HR LECTURE) - ELEMENTS OF CYTOLOGY, CELL CULTURES, CELL VIABILITY ASSAYS (ABOUT 3 HR FRONTAL LECTURE) - METHODS FOR PROTEIN AND DNA PURIFICATION: HOMOGENIZATION OF CELLS AND TISSUES, DIFFERENTIAL PRECIPITATION METHODS, CENTRIFUGATION TECHNIQUES (ABOUT 5 H FRONTAL LECTURE) - CHROMATOGRAPHIC TECHNIQUES (ABOUT 5 H FRONTAL LECTURE) - ELECTROFORETIC METHODS FOR PROTEINS AND NUCLEIC ACIDS (ABOUT 5 H FRONTAL LECTURE) - SPECTROSCOPIC METHODS FOR THE STUDY OF PROTEINS AND NUCLEIC ACIDS: UV-VIS ABSORPTION SPECTROSCOPY, FLUORESCENCE SPECTROSCOPY, CIRCULAR DICHROISM, IR, NMR (ABOUT 4 H FRONTAL LECTURE) - PROTEIN AND NUCLEIC ACID SEQUENCING TECHNIQUES: CLASSICAL AND NEXT-GENERATION TECHNIQUES (ABOUT 4 H FRONTAL LECTURE) - IMMUNOCHEMICAL TECHNIQUES (IMMUNOPRECIPITATION, WESTERN BLOT, ENZYME IMMUNOASSAYS (ABOUT 4 H FRONTAL LECTURE) - RICOMBINANT DNA METHODOLOGIES (HYBRIDIZATION, PCR, CLONING, RECOMBINANT PROTEINS) (ABOUT 4 H FRONTAL LECTURE) - APPROACHES TO THE STUDY OF STRUCTURAL BIOINFORMATICS (APPROXIMATELY 4 H OF GUIDED CLASSROOM EXERCISES) TOPICS COVERED IN BIOCHEMICAL LABORATORY EXERCISES (TOTALING 12 H) ARE: PREPARATION OF PROTEIN EXTRACTS FROM CELL CULTURES, COLORIMETRIC METHODS FOR PROTEIN ASSAY, ELECTROPHORESIS, PCR.

Contents

THE TEACHING INCLUDES 40 HOURS OF DIDACTICS IN THE FORM OF FRONTAL LECTURES AND GUIDED CLASSROOM EXERCISES, AND 12 HOURS OF BIOCHEMISTRY LABORATORY (1 ECTS: BY PROF. GAETANA PAOLELLA), DURING WHICH STUDENTS DIRECTLY TEST WHAT THE LECTURER INTRODUCES IN THE LECTURE (USING A LEARNING-BY-DOING APPROACH).

TOPICS COVERED IN THE CLASSROOM LECTURES ARE:
- STRUCTURAL FEATURES OF PROTEINS AND NUCLEIC ACIDS (APPROXIMATELY 2 HR LECTURE)
- ELEMENTS OF CYTOLOGY, CELL CULTURES, CELL VIABILITY ASSAYS (ABOUT 3 HR FRONTAL LECTURE)
- METHODS FOR PROTEIN AND DNA PURIFICATION: HOMOGENIZATION OF CELLS AND TISSUES, DIFFERENTIAL PRECIPITATION METHODS, CENTRIFUGATION TECHNIQUES (ABOUT 5 H FRONTAL LECTURE)
- CHROMATOGRAPHIC TECHNIQUES (ABOUT 5 H FRONTAL LECTURE)
- ELECTROFORETIC METHODS FOR PROTEINS AND NUCLEIC ACIDS (ABOUT 5 H FRONTAL LECTURE)
- SPECTROSCOPIC METHODS FOR THE STUDY OF PROTEINS AND NUCLEIC ACIDS: UV-VIS ABSORPTION SPECTROSCOPY, FLUORESCENCE SPECTROSCOPY, CIRCULAR DICHROISM, IR, NMR (ABOUT 4 H FRONTAL LECTURE)
- PROTEIN AND NUCLEIC ACID SEQUENCING TECHNIQUES: CLASSICAL AND NEXT-GENERATION TECHNIQUES (ABOUT 4 H FRONTAL LECTURE)
- IMMUNOCHEMICAL TECHNIQUES (IMMUNOPRECIPITATION, WESTERN BLOT, ENZYME IMMUNOASSAYS (ABOUT 4 H FRONTAL LECTURE)
- RICOMBINANT DNA METHODOLOGIES (HYBRIDIZATION, PCR, CLONING, RECOMBINANT PROTEINS) (ABOUT 4 H FRONTAL LECTURE)
- APPROACHES TO THE STUDY OF STRUCTURAL BIOINFORMATICS (APPROXIMATELY 4 H OF GUIDED CLASSROOM EXERCISES)

TOPICS COVERED IN BIOCHEMICAL LABORATORY EXERCISES (TOTALING 12 H) ARE: PREPARATION OF PROTEIN EXTRACTS FROM CELL CULTURES, COLORIMETRIC METHODS FOR PROTEIN ASSAY, ELECTROPHORESIS, PCR.

	Teaching Methods
	TEACHING INVOLVES 52 HOURS (6 CFU) OF DIDACTICS DIVIDED INTO LECTURES (40 HRS), AND PRACTICAL LABORATORY (1 CFU, 12 HRS) BY PROF. PAOLELLA. CLASSROOM LECTURES WILL BE CONDUCTED WITH THE AID OF PROJECTIONS AND ACCESS TO MULTIMEDIA MATERIALS FREELY AVAILABLE ONLINE. GUIDED STRUCTURAL BIOINFORMATICS TUTORIALS WILL INVOLVE GUIDED ACTIVITIES IN WHICH STUDENTS WILL WORK ON COMPUTERS OR OTHER DEVICES THAT ALLOW ONLINE ACCESS AND LEARN WHAT ARE THE MAIN BIOINFORMATICS TOOLS FOR CHARACTERIZING SEQUENCE, STRUCTURE AND FUNCTION OF PROTEINS, AND THEIR APPLICATIONS IN THE BIOLOGICAL FIELD. ACTIVITIES IN THE BIOCHEMICAL LABORATORY WILL BE CARRIED OUT BY PROF. PAOLELLA AND MAY INCLUDE SUDIVISION IN GROUPS OF STUDENTS, ACCORDING TO THE NUMBER OF THE CLASS.

Teaching Methods

TEACHING INVOLVES 52 HOURS (6 CFU) OF DIDACTICS DIVIDED INTO LECTURES (40 HRS), AND PRACTICAL LABORATORY (1 CFU, 12 HRS) BY PROF. PAOLELLA.
CLASSROOM LECTURES WILL BE CONDUCTED WITH THE AID OF PROJECTIONS AND ACCESS TO MULTIMEDIA MATERIALS FREELY AVAILABLE ONLINE. GUIDED STRUCTURAL BIOINFORMATICS TUTORIALS WILL INVOLVE GUIDED ACTIVITIES IN WHICH STUDENTS WILL WORK ON COMPUTERS OR OTHER DEVICES THAT ALLOW ONLINE ACCESS AND LEARN WHAT ARE THE MAIN BIOINFORMATICS TOOLS FOR CHARACTERIZING SEQUENCE, STRUCTURE AND FUNCTION OF PROTEINS, AND THEIR APPLICATIONS IN THE BIOLOGICAL FIELD.
ACTIVITIES IN THE BIOCHEMICAL LABORATORY WILL BE CARRIED OUT BY PROF. PAOLELLA AND MAY INCLUDE SUDIVISION IN GROUPS OF STUDENTS, ACCORDING TO THE NUMBER OF THE CLASS.

	Verification of learning
	THE ACHIEVEMENT OF THE TEACHING OBJECTIVES IS CERTIFIED BY PASSING AN EXAMINATION WITH EVALUATION IN THIRTIETHS. THE EXAMINATION INCLUDES AN ORAL TEST THAT CONSISTS OF AN INTERVIEW WITH QUESTIONS AND DISCUSSION ON THE THEORETICAL AND METHODOLOGICAL CONTENT INDICATED IN THE PROGRAM OF THE TEACHING AND IS AIMED AT ASCERTAINING THE LEVEL OF KNOWLEDGE AND ABILITY OF UNDERSTANDING ACHIEVED BY THE STUDENT, AS WELL AS VERIFYING THE ABILITY TO EXPOSITION USING THE APPROPRIATE TERMINOLOGY AND THE ABILITY TO INDEPENDENTLY ORGANIZE THE EXPOSITION ON THE SAME TOPICS WITH THEORETICAL CONTENT. WILL ALSO BE ASSESSED ALWAYS BY INTERVIEW THE KNOWLEDGE OF THE VARIOUS NOTIONS LEARNED DURING THE HOURS OF PRACTICE AND LABORATORY. THE TEST AS A WHOLE MAKES IT POSSIBLE TO ASCERTAIN BOTH THE ABILITY TO KNOW AND UNDERSTAND, THE ABILITY TO APPLY THE SKILLS ACQUIRED, THE ABILITY TO EXPOUND, AND THE ABILITY TO LEARN AND DEVELOP SOLUTIONS IN AUTONOMOUS JUDGMENT. MINIMUM SCORE (18/30) IS GRANTED WHEN THE STUDENT DEMONSTRATES LIMITED KNOWLEDGE AND ABILITY TO ESTABLISH CONNECTIONS BETWEEN THE TOPICS COVERED. MAXIMUM SCORE (30/30) IS GRANTED WHEN THE STUDENT SHOWS COMPLETE AND APPROPRIATE KNOWLEDGE AND IS ABLE TO ESTABLISH CONNECTIONS BETWEEN THE TOPICS COVERED. HONORS WILL BE AWARDED TO STUDENTS WHO HAVE ANSWERED ALL QUESTIONS CORRECTLY, DEMONSTRATING EXCELLENT LANGUAGE SKILLS AND CRITICAL ABILITY IN EXPOSITION, IN ADDITION TO THE KNOWLEDGE REQUIRED TO ACHIEVE THE HIGHEST MARK.

Verification of learning

THE ACHIEVEMENT OF THE TEACHING OBJECTIVES IS CERTIFIED BY PASSING AN EXAMINATION
WITH EVALUATION IN THIRTIETHS.
THE EXAMINATION INCLUDES AN ORAL TEST THAT CONSISTS OF AN INTERVIEW WITH QUESTIONS AND DISCUSSION ON THE THEORETICAL AND METHODOLOGICAL CONTENT INDICATED IN THE PROGRAM OF THE TEACHING AND IS AIMED AT ASCERTAINING THE LEVEL OF KNOWLEDGE AND ABILITY OF UNDERSTANDING ACHIEVED BY THE STUDENT, AS WELL AS VERIFYING THE ABILITY TO EXPOSITION USING THE APPROPRIATE TERMINOLOGY AND THE ABILITY TO INDEPENDENTLY ORGANIZE THE EXPOSITION ON THE SAME TOPICS WITH THEORETICAL CONTENT. WILL ALSO BE ASSESSED ALWAYS BY INTERVIEW THE KNOWLEDGE OF THE VARIOUS NOTIONS LEARNED DURING THE HOURS OF PRACTICE AND LABORATORY.
THE TEST AS A WHOLE MAKES IT POSSIBLE TO ASCERTAIN BOTH THE ABILITY TO KNOW AND UNDERSTAND, THE ABILITY TO APPLY THE SKILLS ACQUIRED, THE ABILITY TO EXPOUND, AND THE ABILITY TO LEARN AND DEVELOP SOLUTIONS IN AUTONOMOUS JUDGMENT.
MINIMUM SCORE (18/30) IS GRANTED WHEN THE STUDENT DEMONSTRATES LIMITED KNOWLEDGE AND ABILITY TO ESTABLISH CONNECTIONS BETWEEN THE TOPICS COVERED.
MAXIMUM SCORE (30/30) IS GRANTED WHEN THE STUDENT SHOWS COMPLETE AND APPROPRIATE KNOWLEDGE AND IS ABLE TO ESTABLISH CONNECTIONS BETWEEN THE TOPICS COVERED.
HONORS WILL BE AWARDED TO STUDENTS WHO HAVE ANSWERED ALL QUESTIONS CORRECTLY, DEMONSTRATING EXCELLENT LANGUAGE SKILLS AND CRITICAL ABILITY IN EXPOSITION, IN ADDITION TO THE KNOWLEDGE REQUIRED TO ACHIEVE THE HIGHEST MARK.

	Texts
	REFERENCE TEXT: STOPPINI, BELLOTTI: "APPLIED BIOCHEMISTRY". EDISES THE LECTURER WILL INDICATE FROM TIME TO TIME WHICH CHAPTERS AND PARAGRAPHS ARE USEFUL TO CONSULT FOR CONTENT LEARNING OTHER TEXTS FOR FURTHER STUDY: BONACCORSI DI PATTI, CONTESTABILE, DI SALVO: "BIOCHEMICAL METHODOLOGIES". AMBROSIAN PUBLISHING HOUSE. THE LECTURER WILL INDICATE FROM TIME TO TIME WHICH CHAPTERS AND PARAGRAPHS IT IS USEFUL TO CONSULT FOR FURTHER STUDY OF PARTICULAR TOPICS. OTHER MATERIALS: AT THE END OF EACH LECTURE THE LECTURER PROVIDES A BULLETED LIST OF ADDITIONAL MATERIALS CONSULTED, ACCESSIBLE ONLINE, PROVIDING THE LINKS DIRECTLY IN THE SLIDES THAT ARE PROVIDED TO STUDENTS.

Texts

REFERENCE TEXT:
STOPPINI, BELLOTTI: "APPLIED BIOCHEMISTRY". EDISES
THE LECTURER WILL INDICATE FROM TIME TO TIME WHICH CHAPTERS AND PARAGRAPHS ARE USEFUL TO CONSULT FOR CONTENT LEARNING

OTHER TEXTS FOR FURTHER STUDY:
BONACCORSI DI PATTI, CONTESTABILE, DI SALVO: "BIOCHEMICAL METHODOLOGIES". AMBROSIAN PUBLISHING HOUSE.
THE LECTURER WILL INDICATE FROM TIME TO TIME WHICH CHAPTERS AND PARAGRAPHS IT IS USEFUL TO CONSULT FOR FURTHER STUDY OF PARTICULAR TOPICS.

OTHER MATERIALS:
AT THE END OF EACH LECTURE THE LECTURER PROVIDES A BULLETED LIST OF ADDITIONAL MATERIALS CONSULTED, ACCESSIBLE ONLINE, PROVIDING THE LINKS DIRECTLY IN THE SLIDES THAT ARE PROVIDED TO STUDENTS.

	More Information
	THE REGULAR ATTENDANCE AT LECTURES IS STRONGLY RECOMMENDED. FURTHER INFORMATION CAN BE OBTAINED BY CONTACTING THE LECTURER DURING OFFICE HOURS OR BY E-MAIL AMARABOTTI@UNISA.IT OR VIA TEAMS PLATFORM.

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Anna MARABOTTI | APPLIED BIOCHEMISTRY