ChE 172 A. Introduction Biotechnology and Biochemical Engineering.pdf

ChE 172 INTRODUCTION TO BIOCHEMICAL ENGINEERING Lecture A...

ChE 172 INTRODUCTION TO BIOCHEMICAL ENGINEERING Lecture A Prepared By: Catalino G. Alfafara Prepared by CG Alfafara  This material has been reproduced and communicated to you by or on behalf of University of the Philippines pursuant to PART IV: The Law on Copyright of Republic Act (RA) 8293 or the “Intellectual Property Code of the Philippines”. The University does not authorize you to reproduce or communicate this material. The Material may contain works that are subject to copyright protection under RA 8293. Any reproduction and/or communication of the material by you may be subject to copyright infringement and the copyright owners have the right to take legal action against such infringement. Do not remove this notice. Prepared by CG Alfafara  © 2020 Catalino Alfafara This course pack is limited for your personal use. Do not reproduce, distribute, or share any part of this course pack in any website or any medium CHE 172 Lecture A Introduction to Biochemical Engineering and Industrial Bioprocessing Prepared by CG Alfafara  A discussion about biochemical engineering begins with a discussion about BIOTECHNOLOGY Prepared by CG Alfafara  BIOTECHNOLOGY  the practical application of biological agents (living/dead cells or the sub-cellular components) in technically useful operations , either in productive manufacture, services operations or environmental management Prepared by CG Alfafara Biological Agents  Microorganisms (Bacteria, Yeasts, Filamentous Fungi, etc)  Plant/Animal Cells  Sub-cellular components, Enzymes Prepared by CG Alfafara Technically Useful Operations  Manufacture of an economically useful/value added product Single cell protein, pharmaceuticals, industrial chemicals, etc  Waste Treatment/Biodegradation/Bioremediation Prepared by CG Alfafara  BIOTECHNOLOGY IS A BIG INDUSTRY (variety of products) From: Doran, P. (1995). Bioprocess Engineering Principles,Academic Press USA Fermentation Product Typical organism used Approximate World Market (tons/yr) Bulk organics Ethanol Saccharomyces cerevisiae 2 x 107 Acetone/butanol Clostridium acetobutylicum 2 x 106 Biomass Single-cell Candida utilis/ Pseudomonas, 0.5-1 x 106 protein methlotrophus, Baker’s yeast Organic acids Citric Acid Aspergillus niger 2-3 x 106 Lactic Acid Lactobacillus delbrueckii 2 x 105 Amino acids L-glutamate Corynebacterium glutamicum 3 x 106 L-lysine Brevibacterium flavum 3 x 105 L-phenylalanine Prepared by CG Alfafara Corynebacterium glutamicum 2 x 103  BIOTECHNOLOGY IS A BIG INDUSTRY (variety of products) From: Doran, P. (1995). Bioprocess Engineering Principles,Academic Press USA Fermentation Product Typical organism used Approximate World Market (tons/yr) Exocellular polysaccharides xanthan gum Xanthomonas campestris 5 x 103 dextran Leuconostoc mesenteroides small Enzymes proteases Bacillus sp. 6 x 102 amylases Bacillus amyloliquifaciens 4 x 103 pectinases Aspergillus niger 10 Vitamins Vitamin B12 Propionicum shermanii 10 Pigments shikonin Lithospermum erythrohizon (plant cell) 60 kg/yr beta-carotene Blakeslea trispora Prepared by CG Alfafara  BIOTECHNOLOGY IS A BIG INDUSTRY (variety of products) From: Doran, P. (1995). Bioprocess Engineering Principles,Academic Press USA Fermentation Product Typical organism used Approximate World Market (tons/yr) Vaccines tetanus Clostridium tetani < 50 kg/yr hepatitis B Surface antigen in recombinant yeast Therapeutic proteins Insulin Recombinant E. coli Interferon Recombinant E. coli < 20 kg/yr Monoclonal antibodies hybridoma cells < 20 kg/yr Antibiotics penicillins Penicillum chrysogenum 3-4 x 105 cepaholosporins Cephalosporium acremonium 1 x 105 tetracyclines Streptomyces aureofaciens 1 x 105 Prepared by CG Alfafara  BIOTECHNOLOGY REQUIRES INTERDISCIPLINARY COOPERATION Successful application of biotechnology can only be achieved from the integration of several scientific disciplines and technologies including Microbiology/Cell Biology Biochemistry Genetics/Molecular Biology Biochemical or Bioprocess Engineering Prepared by CG Alfafara  Biotechnology: An Interdisciplicary Endeavor The ability to harness capabilites of cells (development of new products and processes) usually starts in the laboratory: Microbiology, Biochemistry , Cell physiology, Molecular biology/Genetics Bringing a “bioprocess” to industrial realization requires engineering skills and know-how: Biochemical engineering or Bioprocess Engineering Prepared by CG Alfafara What is the importance of biochemical engineering in the biotechnology industry? Biochemical engineering is one of the major areas of biotechnology important to its commercialization (Lee 1992). Successful commercialization of biotechnology requires the development of a technologically viable and economically efficient process. Prepared by CG Alfafara Role of the biochemical engineer for commercial realization of biotechnology (1)Bioreactor: scale up, design, optimal operation and control (2)Downstream processing equipment: design and operation (3)Fermentation plant design Prepared by CG Alfafara  ORIGINS AND EVOLUTION OF BIOTECHNOLOGY How did industrial fermentations begin? Prepared by CG Alfafara EVOLUTION OF BIOTECHNOLOGY The First Wave  Microbial Production of Food and Beverages 6000 BC Sumerians and Babylonians were already drinking beer (accidental observation?) 4000 BC Egyptians were already baking leavened bread wine was known in the Near East by the time of the Book of Genesis Prepared by CG Alfafara  ANTIQUITY OF FERMENTATION Relief picture found on the wall of a 5th Dynasty Egyption tomb depicting scenes of baking and brewing (1) Pounding, winnowing, grinding of wheat. (2) Soaking coarse ground flour in water allowing some of whole grains to malt or sprout. (3) Kneading the leavened dough and fashioning to loaves of different sizes, and baking. Baker is portrayed with a characteristic attitude, raking the fire with a long-handled instrument on one hand and shielding his eyes with the other. Prepared by CG Alfafara SOURCE: Scientific American (Industrial Microbiology Edition). September 1981, p. 67  ANTQUITY OF FERMENTATION Relief picture found on the wall of a 5th Dynasty Egyption tomb depicting scenes of baking and brewing. Mash strained into a fermenting vat, which rests on a stand resembling a coiled rope. After fermenting for a few days, finished beer is poured from the vat into pottery jars which are promptly capped, sealed with clay and placed in storage. The Egyptian brewers relied initially on yeasts from the air or on the skin or husk of fruits and cereals; later a pure or almost pure yeast strain became available. Prepared by CG Alfafara SOURCE: Scientific American (Industrial Microbiology Edition). September 1981, p. 67 EVOLUTION OF BIOTECHNOLOGY The First Wave  Microbial Production of Food and Beverages More of a “craft”; involvement of microbes not yet known PHOTO: Scientific American (Industrial Prepared by CG Alfafara Microbiology Edition). September 1981, p. 67  EVOLUTION OF BIOTECHNOLOGY The First Wave  Microbial Production of Food and Beverages By 17th Century, the realization that microorganisms had a http://www.microbeworld.org/images/meetmicros/tools/tool_1_leuw-scope.jpg role in wine, beer making started Anton van Leeuwenhoek’s microscope But…….. http://www.microscopeworld.com/MSWorld/images/bacteria.jpg http://www.delta-education.com/images/products/1937520.jpg http://www.foxnews.com/images/250204/0_61_bacteria_e_coli.jpg  EVOLUTION OF BIOTECHNOLOGY The First Wave  Microbial Production of Food and Beverages Louis Pasteur gave definitive proof of the the fermentative abilities of http://www.fsis.usda.gov/OA/speeches/2004/em_nma/Slide2.JPG microorganisms Swan flask The long "swan-like" neck is open to air, but prevents dust and air- Louis Pasteur can justifiably borne microbes from reaching the liquid. “Some of Pasteur's be considered as the father preparations are at the Pasteur Institute, Paris where they of biotechnology continue to remain sterile for more than 100 years” http://www.labexplorer.com/l ouis_pasteur.htm http://www.wfu.edu/~xiany6/images/1_6.gif EVOLUTION OF BIOTECHNOLOGY The First Wave  Microbial Production Cheese of Food and Beverages Other Microbially bases http://www.dairygoatjournal.com/issu es/07_08_06b.jpg processes: fermented milk, yoghurt, cheeses, soy sauce, tempeh, etc. Yoghurt http://www.oburkedi.com/im/200 6/0814-yogurt.jpg Soy sauce http://www.seasonedpioneers.co.uk/as sets/recipes/Soy%20Sauce.jpg Prepared by CG Alfafara EVOLUTION OF BIOTECHNOLOGY The First Wave Kim-chi  Microbial Production of Food and Beverages http://www.menumagazine.co.uk/archive/feb06/image Other Microbially bases s/kimchi.jpg processes: fermented milk, yoghurt, cheeses, Tempeh soy sauce, tempeh, etc. http://www.zenpawn.com/ve gblog/2006/04/25/homemad e-tempeh/ Natto http://www.sumitomo.gr.jp/english/disco veries/food/images/p_natto_top.jpg Video  How to eat “natto” https://www.youtube.com/watch?v=a9a7LKle9AQ https://www.youtube.com/watch?v=H84OY6F_hQo Prepared by CG Alfafara  EVOLUTION OF BIOTECHNOLOGY The Second Wave Biotechnological processes initially developed under Open Vat Fermentation non-sterile conditions  many industrial compounds such as http://www.ytbioteknik.uu.se/Center ethanol, acetic acid, organic acids, /BPU/services/fermentation/images/ butanol and acetone were produced by 10L2.jpg the end of the 19th century by microbial fermentation procedures Biological that were open to the environment Wastewater  the control of contaminating microorganisms were achieved by Treatment careful manipulation of the http://upload.wikimedia.org/wikipedia/co ecological environment and not by mmons/thumb/1/1b/Trickling_filter_bed _2_w.JPG/250px- complicated engineering practices. Trickling_filter_bed_2_w.JPG  municipal composting of solid wastes and wastewater treatment are outstanding examples of non-sterile Composting biotechnology http://www.rockwa terfarms.com/logo. http://www.wastenot.mb.ca/ne gif w-images/compost/compost3- big.jpg  EVOLUTION OF BIOTECHNOLOGY The Third Wave The introduction of sterility to biotechnological processes A 1940’s: a new direction in biotechnology with the introduction of complicated engineering techniques to the mass cultivation of microorganisms to ensure that the particular biological process could proceed at higher yields with the exclusion of contaminating microorganisms. (birth of biochemical engineering thought to be here) period of increasing volumes of biotechnological activities: antibiotics, Modern Bioreactor amino acids, organic acids, enzymes, http://www.biotopics.co.uk/edexcel/fermtr.gif steroids, polysaccharides, vaccines Prepared by CG Alfafara EVOLUTION OF BIOTECHNOLOGY The Fourth Wave Explosive developments in molecular biology and process control have created new and exciting opportunities to create new frontiers and to improve greatly the efficiency and economics of the established biotechnological industries http://www.brc.riken.jp/lab/epd/Eng/catalog/pcellc.shtml production of human insulin from E. coli, monoclonal antibodies for detection and treatment of diseases plant tissue/ animal cell culture protoplast fusion artificial intelligence for control http://www.selectscience.net/images/products/931_CELLSPIN.jpg of bioreactors artificial organs, stem cells http://www.cebtechservices.com/cellculture2.jpg Prepared by CG Alfafara  THE BIRTH OF BIOCHEMICAL ENGINEERING Prepared by CG Alfafara  The Birth of Biochemical Engineering WORLD WAR II Necessity to produce large quantities of more effective antimicrobials for the war effort (WW II) Many soldiers dying from infectious wounds Sulfa drugs losing effectivity due to resistance http://www.clemson.edu/caah/history/FacultyPages/PamMack/lec122sts/cowan13.html Prepared by CG Alfafara  The Birth of Biochemical Engineering Scientists at Oxford University rediscover Alexander Flemming’s earlier publication on the germicidal properties of http://users.r “mold juice” which was largely unnoticed cn.com/jkimb all.ma.ultran et/BiologyPa Germicidal component ges/P/Penicil lium.jpg named “Penicillin”, after the genus of the mold Oxford University scientists proved that http://en.wikipedia.org/wiki/Alexander_Fleming penicillin could effectively treat wound infections Prepared by CG Alfafara The Birth of Biochemical Engineering Penicillin was heralded as a “wonder drug” at the time Intial attempts for mass production was space consuming with very low product yields (as low as 0.001 g/L) Prepared by CG Alfafara The Birth of Biochemical Engineering Pfizer was the first pharmaceutical company to take the challenge of mass producing penicillin After 3 years of difficulty (low yield and stability of http://www.biotopics.co.uk/microbes/ferm.gif product), and new approach using deep tank fermentation (“submerged fermentation”) was attempted. http://www.pfizer.com/pfizer/history/images/tank.gif The Birth of Biochemical Engineering The chemical engineering techiques learned for high penicillin production by fermentation in a stirred tank reactor became the foundation for the biochemical engineering field The penicillin yield increased 50-fold Prepared by CG Alfafara  The Birth of Biochemical Engineering The penicillin process also established a paradigm for bioprocess development and biochemical engineering. This paradigm still guides much of the profession’s thinking The mind set of bioprocess engineers was cast with the penicillin experience Prepared by CG Alfafara What is the importance of biochemical engineering in the biotechnology industry? Biochemical engineering is one of the major areas of biotechnology important to its commercialization (Lee 1992). Successful commercialization of biotechnology requires the development of a technologically viable and economically efficient process. Prepared by CG Alfafara Commercialization of Biotechnology Involves Scale Up from Laboratory Scale (small) to Production Scale (large) Prepared by CG Alfafara  Small Scale vs Large Scale How Does the Story Change? SMALL SCALE PHOTO: Scientific American LARGE (Industrial Microbiology Prepared by CG Alfafara SCALE Edition). September 1981, p. 67  Scale Up Comics Prepared by CG Alfafara http://www.av.fh-koeln.de/professoren/forschung/ricckmann-forschung.html  Small Scale vs Large Scale How does the story change when you implement the following stages of fermentation from small to large scale? (1) Medium preparation (2) Sterilization (3) Inoculation (4) Main Fermentation (5) Product Separation and Purification (downstream processing) Prepared by CG Alfafara Medium Preparation LAB SCALE LARGE SCALE medium components Medium components weighed place in flask, weighed, put into dissolved in water by bioreactor, dissolved in stirring rod water by agitation system * biochemical engineer calculates the power requirements of agitator motor (too low:inefficient mixing, too high: Prepared by CG Alfafara costly) Sterilization LAB SCALE LARGE SCALE Autoclave (steam), 121 Steam injection into oC, 10-15 minutes bioreactor jacket * biochemical engineer calculates steam requirements to achieve sterilization Prepared by CG Alfafara Sterilization LARGE SCALE Steam injection into bioreactor jacket - biochemical engineer calculates steam requirements to Steam achieve sterilization out Prepared by CG Alfafara http://www.biotopics.co.uk/microbes/ferm.gif Steam in Inoculation LAB SCALE LARGE SCALE wire loop progressive inoculation sterile pipets using seed fermentors biochemical engineering “rule of thumb” for inoculation size: 3% to 10% of reactor working volume Prepared by CG Alfafara Inoculation LAB SCALE wire loop Sterile pipets Prepared by CG Alfafara  LARGE SCALE progressive inoculation using seed fermentors Inoculation biochemical engineering “rule of thumb” for inoculation size: 3% to 10% of reactor working volume slant Seed Fermentors flask Main Fermentor http://www.visitmonmouth.com/health/labpages/ImagesLab/entero3.png http://www.niroinc.com/html/gea_liquid_processing/Microorganism_cell_fermentation_systems.htm Prepared by CG Alfafara  LARGE SCALE progressive inoculation using seed fermentors Inoculation biochemical engineering “rule of thumb” for inoculation size: 3% to 10% of reactor working volume Inoculating a small to medium-sized fermenter Prepared by CG Alfafara Fermentation LAB SCALE LARGE SCALE flasks, petri plates, small Large scale bioreactors fermenters * biochemical engineers, determine power requirements for agitation equipments, cooling water requirements, aeration rates (for aerobic cultvtn) Prepared by CG Alfafara Fermentation LARGE SCALE Large scale bioreactors - biochemical engineers, determine power requirements for agitation equipments, cooling water requirements, aeration rates (for aerobic cultvtn), management of exhaust gases, etc PHOTO: Scientific American (Industrial Microbiology Edition). September 1981, p. 67 Prepared by CG Alfafara Downstream processing LAB SCALE LARGE SCALE Lab scale centrifuges, Large equipment for filters, chromatography product separation and equipment, etc purification Prepared by CG Alfafara Downstream LARGE SCALE Large equipment for product processing separation and purification Cell Separation: centrifuge http://www.marstechusa.com/discbowl.html Product separation/purification equipment Chromatography columns Filters, etc Prepared by CG Alfafara http://www.pharmaceutical-technology.com/contractor_images/polypeptide/hplc.jpg