ENZYME TECHNOLOGY (2014/2015, Semester 2) 

 MODULE OUTLINE Created: 07-Jan-2015, Updated: 07-Jan-2015
Module Code CN4247R
Semester Semester 2, 2014/2015
Modular Credits 4
Faculty Engineering
Department Chemical & Biomolecular Engineering
Timetable Timetable/Teaching Staff
Module Facilitators
ASSOC PROF Li Zhi Lecturer
Tags --

Learning Outcomes | Prerequisites | Teaching Modes | Schedule | Syllabus | Assessment | Workload

The aim of this model is to provide students with knowledge on enzyme technologies for industrial production of chemicals via biotransformation. This module will start with general introduction about enzyme, enzymatic transformation, and enzymatic process. It will be followed by various components in the development of an enzymatic process: enzyme classes and enzymatic reactions; enzyme discovery and high-throughput screening and detection methods; enzyme purification, characterization, structure, function, and selectivity; protein engineering; cell engineering; biotransformation with isolated enzymes and microbial cells; reaction engineering; enzyme in organic solvent, two-liquid phase system, and enzyme stabilization; cofactor regeneration; and product recovery. Finally, the students will learn process economics and industrial examples on the enzymatic production of fine chemicals.

Module Learning Outcomes:
On successful completion of this module, students will be able to:

  1. Explain different types of enzyme-catalysed transformations;
  2. Develop suitable methods for enzyme discovery, engineering and purification;
  3. Analyze enzyme activity, kinetics, and enantioselectivity, and evaluate cost of bioproduct manufacture;
  4. Design biotransformations with isolated enzyme or whole cells as catalysts;
  5. Investigate problems on biocatalytic process development.

CN2116, LSM1401

 This module consists of 3 h Lecture and 1 h Tutorial per week.

Week Mon. (9:00-11:00)
Tue. (9:00-10:00)
Tue. (10:00-11:00)
Week 1 Lecture Lecture -
Week 2 Lecture Lecture Tutorial
Week 3 Lecture Lecture Tutorial
Week 4 Lecture Lecture Tutorial
Week 5 Lecture Lecture Tutorial
Week 6 Lecture Test 1 Tutorial
Recess Week - - -
Week 7 Lecture Lecture Tutorial
Week 8 eLearning eLearning eLearning
Week 9 Lecture Lecture Tutorial
Week 10 Lecture Lecture Tutorial
Week 11 Lecture Test  2 Tutorial
Week 12 Lecture Lecture Student Presentation
Week 13 Lecture Student Presentation Tutorial

Introduction (3 hours)
Definition and history of enzyme and enzyme catalysis, prejudices and advantages of enzyme catalysis, chemo-, regio-, and enantioselectivity,  biotransformation with isolated enzyme and whole cells, industrial application, enzyme assay; enzyme activity, whole cell activity, space-time-yield.
Enzyme classes and reactions (3 hours)
Enzyme classification (EC numbers), oxidoreductase, co-factor, transferase, hydrolase, lyase, isomerase, examples of enzymatic reactions.
Enzyme selection (3 hours)
Target reaction definition and selection, source of enzymes, enzyme or microorganism screening strategies, microtiter plate, high-throughput screening, detection methods, enantioselectivity assays.
Enzyme purification and characterization (3 hours)
Enzyme purification techniques and general principles: SEC, IEC, HIC, IMAC, affinity matrices, affinity tags, and fusion proteins, examples. Enzyme characterization methods: molecular mass, IEP, multimers, amino terminal sequence, structure determination.
Enzyme biochemistry and enantioselectivity (3 hours)
Michaelis-Menten kinetics, kinetic constants Kcat, Km, inhibition, enzyme selectivity, enantioselectivity, kinetic resolution vs dynamic resolution.
Enzyme engineering (5 hours)
Physicochemical classification of amino acids, replacement strategy, Dayhoff matrix, in vivo and in vitro mutagenesis (random, semi random), site directed mutagenesis, sequence comparisons and predictions, directed evolution, DNA shuffling, examples.
Biotransformation with whole cells (3 hours)
Cell growth, whole cells biotransformation, recombinant microorganisms, Multi-enzyme reactions, secondary metabolites, fermentation.
Biotransformation with enzymes (3 hours)
Cofactor regeneration systems (NADPH, NADH, ATP), coupled substrate approach, coupled enzyme approach, permeabilized cells, enzyme stabilization, immobilization technology, membrane reactors.
Biotransformation in organic solvent (4 hours)
Enzymes and solvents: solubility of enzymes, substrates and products, water activity. organic solvents: ‘Klibanov’ rules, reaction reversal, ‘memory’ effect. Ion-paired enzyme. Enzyme catalysis in ionic liquid.
Product recovery (3 hours)
Product recovery strategies, intracellular and extra-cellular compounds, disruption of cells, separation of cells, primary isolation (solvent extraction, sorption, adsorption, precipitation, membrane filtration, reverse osmosis, pervaporation, perstraction), further purification.
Process economics (3 hours)
Process design, flow sheets, calculations, stringent and relaxed parameters, process evaluation, quantitation, economics of a process, research choices based on process cost considerations.
Biotransformation in industry (3 hours)
Case studies based on operational industrial processes.  

CA: 50% [Test 1, 15%; Test 2, 15%; Term paper, 10%; Presentation (a selected journal paper), 10%]
Final exam: 50%


Workload Components : A-B-C-D-E
A: no. of lecture hours per week
B: no. of tutorial hours per week
C: no. of lab hours per week
D: no. of hours for projects, assignments, fieldwork etc per week
E: no. of hours for preparatory work by a student per week