Chapter1Introduction

1、Why? What is the most expensive or dearest substance in the world? Update gold price = $1103/ounce=$35.45/g= $ 1.32 million per carats= $ 1.32 million per carats $6.6 million per gram $6.6 million per gramAbout 682 times!It is difficult to separate them:(2) Complex components （成分复杂）(1) desired produ

2、cts in dilute suspension（目标产物浓度低）(3) Biological products require high quality (生物产品高质量)： * purity（纯度） * sanitation（卫生） * biological activity（生物活性）How to solve it?What is Bioseparations? (1) A collective term Includes all the steps or methods required in order to recover useful biochemicals or biopro

3、ducts from any kind of industrial processes. ( (所有从任何工业过程中回收有用的生物化学品或生物产品的所有从任何工业过程中回收有用的生物化学品或生物产品的方法、步骤方法、步骤) ) Biochemicals or Bioproducts chemical substances or combinations of chemical substances that are made by living things range from methanol (甲醇) to whole cells. Extracted from whole plants

4、 and animals (提取) synthesized in bioreactors containing cells or enzymes (生物合成)What is Bioseparations(2) Downstream processing for biotechnology What is BioseparationsWhat is biotechnology? What is BioseparationsThe application means the use of carefully cultured microorganisms, animal cells and pla

5、nt cells to produce products useful to humans. （即指利用培养微生物、动植物细胞来生产对人有用的产品。） What is Bioseparations What is BioseparationsAbove all: Biosepartions is a fraction of biotechnology; It consists of a series of steps (or processes);c. Its objective is to obtain ( recover) useful biochemicals or bioproduct

6、s What is BioseparationsUpstream ProcessingbiosynthesisBioseparation Downstreamprocessingdownstream processWhat is BioseparationsBioseparationBioproductsVariety ( structures and sizes )Lability ( temperature, pH, concentration) More unit operationsWhat is BioseparationsThe characteristics of Biosepa

7、rationThe characteristics of BioseparationAmino Acid and their AbbreviationsThe characteristics of BioseparationSmall molecules Primary metabolitesSecondary metabolitesThe characteristics of BioseparationProteinsThe characteristics of BioseparationThe characteristics of BioseparationProteins腺嘌呤鸟嘌呤胞嘧

8、啶尿嘧啶胸腺嘧啶The characteristics of BioseparationThe characteristics of BioseparationThe characteristics of Bioseparation low concentration of the targets (目标产物低浓度目标产物低浓度)The concentration of the targets is very low in fermentation liquor. For example, penicillin: 4.2%, gentamycin(庆大霉素庆大霉素): 0.2%. Moreov

9、er, the concentration of the targets in some animals tissue culture is only about 550 g/ml. Biological (or chemical) activity (活性活性) Bioproducts are sold for their biological (or chemical) activity:Methanol: solvent activity; ethanol: neurological activity; penicillin: antibacterial activity; Taxol

10、(红红豆杉醇豆杉醇): anticancer activity, and so on. The characteristics of BioseparationThe characteristics of Bioseparation Fermentation broth being a complicated mixture (complex components) (发酵液成分复杂发酵液成分复杂)The fermentation broth is the complicated mixture of many components( soluble or insoluble), not on

11、ly including macromolecules such as proteins, nucleic acid, polysaccharides but also including intact organisms, fragmented mycelia, other insoluble fractions of medium ingredients, residual substrates, etc. The characteristics of Bioseparation Poor stability ( (稳定性差稳定性差) ) Due to chemical degradati

12、on or microbial degradation, the products can be inactivated by temperature, pH or various enzymes. High purity and strict sanitation criterion Almost all products are used as medicine, biological reagents or food, their purity must meet the need of pharmacopoeia (药典药典), reagent criterion or food cr

13、iterion etc. All above makes bioseparations much more difficult than chemical separations!The characteristics of Bioseparation The diversity of products spawns (results in) the broad spectrum of separation methods used(2) Characteristics of Bioseparations Broad spectrum of separation methods （产品的多样性

14、导致分离方法的多样性）（产品的多样性导致分离方法的多样性） most bioseparation methods come from separations of chemicals ( (大多数生物分离方法来源于化学品的分离方法大多数生物分离方法来源于化学品的分离方法) )The characteristics of Bioseparation(2) Characteristics of Bioseparations This table shows that 80% of all the separation methods for conventional chemicals are p

15、ractical in biotechnology. (大约大约80%80%的化工分离方法可应用于生物分离技术的化工分离方法可应用于生物分离技术) The characteristics of Bioseparation These separations are elaborate and expensive Both dilute feed and products with high purity make the separations be elaborate and expensive.More equipment and labor than all other parts of

16、 the process combined are needed.Costing a substantial fraction of the total cost of the final product. This fraction can be as high as 90% or more!The characteristics of Bioseparation Yeast biomass, penicillin G, and certain enzyme processing costs are percentages of the selling prices which are 20

17、%, 20-30% and 60-70% respectively. Products expressed by genetic engineering, their processing costs can be as high as about 8590% of their selling prices. The processing cost of asparaginase (天冬酰胺酶) used as pharmaceuticals is about 85% of its selling prices because of its high purity. The character

18、istics of BioseparationPrice vs Production红细胞生成素红细胞生成素长春碱长春碱红豆杉醇红豆杉醇洋地黄洋地黄果胶酶果胶酶The characteristics of Bioseparation The recovery and the purification processes must be well conceived and well designed!Important! Organic molecules, $5 - $10 / g Biopolymers, $1 - $5 / g Food & beverage additives,

19、 $20,000 / g How to lower them?（因此必须仔细考虑和设计产品的回收和纯化过程）（因此必须仔细考虑和设计产品的回收和纯化过程）The characteristics of BioseparationSo, Before designing, what we should ask and answer are:What is the value of the product? ( (产品价值产品价值) )What is an acceptable product quality? ( (产品质量产品质量) )Where is the product and impur

20、ities in each process stream? ( (产物及杂质在加工过程中出现位置产物及杂质在加工过程中出现位置) )The characteristics of BioseparationWhat are the unusual physicochemical properties of the product and the principal impurities? ( (产物及主要杂质独特的物化性质是什么？产物及主要杂质独特的物化性质是什么？) )What are the economics of various alternative separations? (各种分

21、离方法的经济性比较各种分离方法的经济性比较) Careful consideration of these questions can help us to determine the optimal separating processes.The characteristics of BioseparationImprove the efficiency of bioseperationReduce the steps of operationsReduce the time of operation processReduce costImprove purityKeep activit

22、yThe characteristics of BioseparationAbove all:这些分离过程不这些分离过程不具备特异性，只具备特异性，只是进行初分是进行初分具有产物的高具有产物的高选择性和杂质选择性和杂质的去除性的去除性Downstream processes for production of antibioticProduction of recombinant human alpha-1-antitrypsin (抗胰蛋白酶抗胰蛋白酶)Recovery of an extracellular enzymeNutrient mediumFermentationMicroorg

23、anismsSelf-cleaning clarifierCell massSelf-cleaning clarifierResidual solidsConcentratione.g. ultrafiltration or vacuum evaporationPrecipitationFilter pressDryingEnzyme powderLipidSterilizationLiquid enzymeRecovery of intracellular enzymesNutritive mediumFermentationMicroorganismsSelf-cleaning clari

24、fier (cell harvesting)Cell disintegrationDilution (if possible)Continuous liquid-liquid Cross-flow extraction with aqueous two-phase systemsSelf-cleaning clarifier (separation of cell debris)Concentration and enrichmentPurificationConcentration/ precipitationBioreactor fluid with cellsChromatography

25、Cell disruptionCentrifugal extractionDialysis Cleaning pro insulinPrecipitation Dialysis Sulfonation Extraction Precipitation Chromatography Cell debrissupernatantB-ChainA-ChainRecovery and purification of human insulinBioreactor fluid pretreatmentSterile ultrafiltrationFiltration or centrifugationT

26、wo-stage ultrafiltrationUltrafiltrationPrecipitation Polishing ultrafiltrationRotary filter Formulation DryingFormulation CellsFilter aidprecoatLiquids or solidsCool to 4-5Filter aidLiquid productSolid productDownstream processing for production of liquid or solid proteaseBioreactor fluid with cellsFiltration Extraction pH2-3Back extractionFiltration Crystallization Ion exchangecentrifugationDrying Kerosene and tertiary amineWater