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The Role of Bioprocess Engineering in Biotechnology. Author: Michael Ladisch. Bioprocess engineering is the discipline that puts biotechnology to work. Biotechnology involves using organisms, tissues, cells, or their molecular components 1 to act on living things and 2 to intervene in the workings of cells or the molecular components of cells, including their genetic material NRC, Biotechnology evolved as a means of producing food, beverages, and medicines.
More than 8, years ago, it was used to make leavened bread. Some 5, years ago, moldy soybean curd was used to treat skin infections in China.
The malting of barley and fermentation of beer was used in Egypt in BC Ladisch, Biology is central to biotechnology. Louis Pasteur proved in that yeast is a living cell that ferments sugar to alcohol; in , he showed that some bacteria kill anthrax bacilli. In , Banting and Best showed that insulin from animals could be used to treat people suffering from diabetes. In , Alexander Fleming showed that growing colonies of Penicillium notatum inhibit Staphylococcus cultures.
They realized that producing Penicillium on a large scale would require isolation and purification procedures that minimized product loss. Early bioprocess engineers found solutions to this problem Aiba et al.
During World War II, government incentives encouraged several pharmaceutical companies to develop cost-effective manufacturing processes for penicillin Hacking, Chemical engineers, industrial chemists, and microbiologists quickly devised methods of countercurrent extraction, crystallization, and lyophilization to recover penicillin in an active, stable form and established the viability of submerged fermentations Matales, The benefits of biotechnology might be an anomaly if it were not for engineering, specifically bioprocess engineering, the discipline that puts biotechnology to work NRC, The realization of the benefits of penicillin required the development of methods of transforming microbial growth on the surface of a moldy cantaloupe to cultures grown in large stirred tanks fed by sterile air Aiba et al.
It took engineers to design the tanks, impellers, pumps, compressors, columns, pipes, and valves that have made biotechnology products available to large numbers of people. The lifesaving benefits of insulin required engineering for the extraction and purification of insulin from cow and pig pancreas, and later, the large-scale propagation of bacteria engineered to make human insulin, as well as methods of recovery, refolding, and purification to obtain an active molecule Ladisch, Biochemical manufacturing and bioseparations have made it possible to purify products derived from biotechnology on a large scale.
High-Fructose Corn Syrup and Bioethanol In , scientists at USDA reported the discovery of an enzyme with the amazing ability to transform glucose to fructose although it required arsenic as a cofactor. In , a version of this glucose isomerase enzyme that did not require arsenate was discovered in a species of Streptomyces. Once it was possible to grow this organism using corn-steep liquor to produce a thermally stable enzyme in a cost-effective way, sugars from corn with sweetness similar to sugar from sugar cane became feasible.
Glucose isomerase which also transformed xylose to xylulose was used to generate the first commercial shipment of corn syrup containing 42 percent fructose in Bioprocess engineers invented systems of fixed beds of the glucose isomerase enzyme and demonstrated the utility of biocatalysts for the large-scale industrial production of biochemicals. They also adapted industrial-scale liquid-chromatography separations used in the petrochemical industry to enrich the fructose content in corn syrup from 42 percent to 55 percent UOP Sarex process , creating percent high fructose corn syrup HFCS.
The HFCS industry grew quickly, particularly after , when patent coverage for using xylose glucose isomerase to convert glucose to fructose was lost due to a civil action suit described in Ladisch, In , yields of corn were 85 bushels per acre; by , they had jumped to bushels per acre. Today, yields range from about to bushels per acre. The wet mills that produced HFCS had the infrastructure, integrated processing, biotechnology, and bioprocess engineering expertise to make million-gallon fermenters conceivable.
They also had access to glucose from corn to fill these tanks with substrates for the production of fuel ethanol, which was introduced in the s NRC, The 81 million acres of corn planted in will provide renewable raw materials, not only to make sugar, but also to make fuel ethanol and other bioproducts, such as monomers and biodegradable plastics.
DNA, Genetic Engineering, and the Biotechnology Industry In , Watson and Crick showed that DNA consists of a double helix with a code of triplets of nucleotides that correspond to specific amino acids and the sequence in which they were assembled. Genes in chromosomes were mapped to the genetic bases of diseases. Cellular processing of DNA and other nucleotides derived from it were beginning to be understood.
A breakthrough came in when Smith et al. Restriction enzymes were then used to cut plasmids circular DNA found in bacteria in a way that allowed scientists to insert new genes. By , Cohen et al. One widely used plasmid is PBR Its 4, nucleotides were completely sequenced in PBR contains unique restriction sites and has genes for antibiotic resistance that allows for the selection of transformed bacteria.
Thus, specific regions restriction sites cleaved by specific restriction enzymes could be identified and foreign genes inserted. In other words, the engineering of genes, or genetic engineering, became possible. When the reconstructed plasmid was reintroduced into a microorganism, E.
PBR carried instructions genes for making enzymes that rendered the antibiotics ampicillin and tetracycline harmless Old and Primrose, Only about 1 percent of E. Cells that contained the plasmid had built-in resistance to the antibiotic; cells that did not contain the plasmid were killed. Thus, biologists had found a way to select for transformed cells.
The stage was now set for the first human protein, human insulin, to be produced in E. In , separate insulin A and B chains were achieved in E. Later, human insulin was produced as a preprohormone, using one fermentation instead of two Chance et al. Eli Lilly licensed the technology and quickly developed the process, and the first recombinant product, human insulin, was marketed in By , human insulin provided an estimated 70 percent of the demand for insulin in the United States.
The production of human insulin required 31 major processing steps, 27 of which are associated with product recovery and purification Prouty, Bioprocess and bioseparation engineering, which provided technology for carrying out complex, biological processes on a large scale, were critical in bringing human insulin to market. Monoclonal Antibodies In , Kohler and Milstein reported that hybrid cells derived from mouse B lymphocytes which secrete antibodies fused to mouse myeloma malignant cells will grow in submerged cultures.
The fused cells, called hybrid myelomas, or hybridomas NRC, , had the capability of growing and dividing, and hence producing, monoclonal antibodies in cell culture. The cells derived from the founder cell are identical to it and produce the same antibodies, which are referred to as monoclonals.
Here was an example of living things acting on livings things and with each other to make a part of a living thing an antibody that had therapeutic uses. Initially, monoclonal antibodies were considered tools for detecting or diagnosing pathogenic microorganisms or cancer cells because of their ability to bind specifically to protein biomarkers that label these cells.
When monoclonal antibodies were linked to toxins to deliver them specifically to cancer cells and other therapeutic uses were discovered, demand for their manufacture increased dramatically.
Bioprocess engineers are working to scale up processes of cell culture to enable manufacturing facilities to meet that demand.
Biopharmaceuticals and Bioproducts Biopharmaceuticals biological molecules with medicinal value include treatments for cancer, heart disease, and autoimmune diseases. Bioproducts are commodity-scale products that often have a lower molecular weight e. Other types of future bioproducts might include functional foods that improve nutrition or contain edible vaccines, biomaterials for paints and coatings, and optical-holographic high-density memories NRC, Bioprocess engineering puts biotechnology to work by providing manufacturing systems to generate bioproducts in large volume, at low cost, and with acceptable purity.
The grand challenge of sequencing the human genome required that many existing bioprocessing tools—fermentation, enzymology, and bioseparations—be mapped onto new biotechnologies—cloning, polymerase chain reaction PCR , and automation of DNA sequence analysis—and used with information-age tools that connected computers through the Internet.
The goal was to generate sequences the order of nucleic acids in DNA and piece them together to discover genes and the nature of information stored in DNA.
To produce enough genetic material, DNA was propagated in microbial cells using bacterial artificial chromosomes. The human genome consists of chromosomes made of DNA associated with a protein that wraps around it to protect it from the effects of mechanical forces.
First, the DNA had to be separated from this protein so that it could be disassembled one nucleic acid at a time. Restriction enzymes were used to break up DNA into small fragments of to base pairs. The fragments were then sequenced and the sequences compared to find overlapping fragments. Computers were used to reassemble the sequences of these overlapping fragments into the original DNA code.
By replicating the procedure in different laboratories, comparing the results against existing databases, and using the Internet and computers to make comparisons, the sequencing task was completed by , several years ahead of schedule. It took another year to determine the number of genes in the human genome now believed to be about 30, but once thought to be , The engineering of automated instruments and software for analyzing nucleic acid sequencing played a major role in achieving this milestone.
The enzyme first isolated from microbes growing in hot springs enables researchers to make millions of copies of a DNA fragment in as little as an hour, enough to sequence or identify the DNA sequence. Biotechnology and Agriculture In the past, plant and animal breeding were used to enhance agriculture by taking advantage of the natural variability of characteristics or inducing mutations or using natural mutations in genes.
Today, genes from other species can be engineered into plants or animals. For example, antiworm protein from the bacterium Bacillus thuringiensis can be engineered into corn or cotton, thus reducing the need for pesticides. Biotechnology has provided the tools for engineering crops resistant to pests or herbicides and animals capable of producing therapeutic proteins.
Another example is sheep that produce human antibodies that can boost the immune system. Organisms with genes transferred from one species to another are called transgenic.
The tools of molecular biology, described in this paper, which were developed largely for medical applications, are being used for cloning genes into microorganisms to produce bioenergy and bioproducts, as well as for the study and modification of metabolic pathways in microorganisms. These studies require gene sequencing and relating protein function to its structure, much as pharmaceutical properties are related to protein structure in the development of new pharmaceuticals.
There is a major difference, however. Outputs of bioenergy such as ethanol will be measured in tons per day, whereas outputs of biopharmaceuticals may be measured in kilograms per year. A yeast has been engineered to produce ethanol from xylose, including the formation of xylulose by xylose isomerase an enzyme that also isomerizes glucose to fructose.
The xylulose then enters an ethanol-producing pathway. Glucose has been fermented to ethanol for millions of years, but when xylose isomerase and other enzymes are cloned into yeast, both glucose and xylose can be fermented. Similarly, a genetically engineered E. When combined with a suite of other bioprocessing technologies, pentose fermentation increases the yield of ethanol from plant materials by 50 percent, moving us a step closer to the transformation of renewable, agricultural residues into fuel-grade ethanol.
Imagine the potential in the United States alone, where an estimated 20 million tons of residues could produce 1. The goal of metabolic-pathway engineering is to understand, and ultimately direct, metabolic pathways in microbial cells to make value-added bioproducts.
Some transgenic animals and plants have been engineered to produce therapeutically important proteins although these are not yet commercial. Agriculture could become a producer of large volumes of therapeutic compounds using small amounts of land.
Engineering will play an important role in the extraction, recovery, and purification of these products.
Biochemical Engineering and Biotechnology, 2nd Edition, outlines the principles of biochemical processes and explains their use in the manufacturing of every day products. The author uses a diirect approach that should be very useful for students in following the concepts and practical applications. This book is unique in having many solved problems, case studies, examples and demonstrations of detailed experiments, with simple design equations and required calculations. The book is appropriate as a college and university text book for undergraduate senior courses and postgraduate course. Students and research scientists in biochemical engineering and biological sciences will find this reference particularly useful for gaining an overview of the subject and planning research activities. It is also useful for research institutes and postgraduates who are involved in practical research in biochemical engineering and biotechnology.
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The Role of Bioprocess Engineering in Biotechnology.
The author uses a diirect approach that should be very useful for students in following the concepts and practical applications. This book is unique in having many solved problems, case studies, examples and demonstrations of detailed experiments, with simple design equations and required calculations. Extensive application of bioprocesses has generated an expansion in biotechnological knowledge, generated by the application of biochemical engineering to biotechnology. Microorganisms produce alcohols and acetone that are used in industrial processes. The knowledge related to industrial microbiology has been revolutionized by the ability of genetically engineered cells to make many new products. Genetic engineering and gene mounting has been developed to enhance industrial fermentation.
Not a MyNAP member yet? Register for a free account to start saving and receiving special member only perks. Research activities and the training of bioprocess engineers for the next decade should be broad enough to enable staffing of bioprocess research, development, and manufacturing functions for biotherapeutics and other classes of bioproducts, including intermediate-value products obtained from renewable resources through bioprocessing, value-added agricultural materials, and waste-processing products and services. This chapter treats elements of bioprocess engineering that must be addressed to meet the needs of industry and the goal of commercializing biotechnology. The principles, culture, and techniques of scientists biologists and chemists are often different from those of bioprocess engineers. The differences can place unnecessary limits on collaboration among members of a bioprocessing-development team and thereby delay engineering considerations to the later stages of bioprocess development.
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