1Biochemical Engineering

Biochemical engineering is a branch of chemical engineering thatmainly deals with the design and construction of unit processes that involve biological organisms of molecules.

Applications of biochemical engineering are used in the food, feed, pharmaceutical, biotechnology and water treatment industries. Will include a study of bioreactors including bioreactor design.

Marta HenriquesESAC-IPC

Cells & nutrients

Biochemical Engineering13-14

Marta HenriquesESAC-IPC

2Cells composition

Water (80%) Dry weight (10-20%) Protein (50%) ex. enzymes Lipids (5-15%) ex. some cells accumulate PHB up to

90%

Others (35-45%) ex. carbohydrates, minerals

Intracellular composition depends on the type and age of the cells and the composition of the nutrients media.

3 The qualitative and quantitative nutritional requirements of cells need to be determined to optimize growth and products formation

Cell Nutrients

Nutrients required by cells can be classified in two categories:

- are needed in concentrations larger than 10-4 M.

C, N, O, H, S, P, Mg 2+, and K+.

- are needed in concentrations less than 10-4 M.

Mo, Zn, Cu, Mn, Ca, Na, vitamins, growth hormones and metabolic precursors.

4Cell Nutrients- Macronutrientscompounds are the major sources of cellular

carbon and energy.

Heterotrophs use organic carbon sources such as carbohydrates, lipid, hydrocarbon as a carbon source.

Autotrophs can use CO2 as a carbon source. They can form carbohydrate through light or chemical oxidation.

In aerobic fermentations, about 50% of substrate carbon is incorporated into cell mass and about 50% of it is used as energy sources.

In anaerobic fermentation, a large fraction of substrate carbon is converted to products and a smaller fraction is converted to cell mass (less than 30%).

Cell Nutrients- Macronutrients

Carbon sources:- In industrial fermentation, the most common carbon sources are molasses (sucrose), starch (glucose, dextrin), corn syrup, and waste sulfite liquor (glucose).

- In laboratory fermentations, glucose, sucrose and fructose are the most common carbon sources. Ethanol, methanol and methane also constitute cheap carbon sources.

5Cell Nutrients- Macronutrients

compounds are important sources for synthesizing protein, nucleic acid.

Nitrogen constitutes 10% to 14% of cell dry weight.

Is incorporated into cell in the form of proteins and nucleic acids.

The most commonly used nitrogen sources are ammonia or ammonium salts such as ammonium chloride, sulfate and nitrate, protein, peptides, and amino acids. Urea can be cheap source.

Some m.o. such as Azotobacter sp. and cyanobacteriafix nitrogen from atmosphere to form ammonium.

6Cell Nutrients- Macronutrients

constitutes about 20% of the cell dry weight.

- Molecular oxygen is required as terminal electron acceptor in the aerobic metabolism of carbon compounds.

- Gaseous oxygen is introduced into growth media by sparging air or by surface aeration.

- Improving the mass transfer of oxygen in a bioreactor is a challenge in reactor control.

Cell Nutrients- Macronutrients

: 8% of dry cell weight

major source: carbohydrates.

- methanogen bacteria can utilize hydrogen as an energy source

: 3% of cell dry weight

- present in nucleic acids and in the cell wall of some gram-positive bacteria.

- a key element in the regulation of cell metabolism.

- sources: Inorganic phosphates.

The phosphate level should be less than 1 mM for the formation of many secondary metabolites such as antibiotics.

7Cell Nutrients- Macronutrients

: 1% of cell dry weight- present in protein and some coenzymes. - source: ammonium sulfate, Sulfur containing aminoacids, cysteine- some autotrophs can use S0 and S2+ as energy sources.

: a cofactor for some enzyme and is required in carbohydrate metabolism. cofactor: any of various organic or inorganic substances necessary to the function of an enzyme. - source: potassium phosphates.

: a cofactor for some enzyme and is present in cell walls and membranes. Ribosomes specifically requires Mg2+ .- sources: Magnesium sulfate or chloride

8Cell Nutrients- Micronutrients

Micronutrients could be classified into the following categories (required less than 10-4 M):

Cell Nutrients- Micronutrients

1. most widely needed elements are Fe, Zn and Mn. Such elements are cofactors for some enzyme and regulate the metabolism.

2. trace elements needed under specific growth conditions are Cu, Co, Mo, Ca, Na, Cl, Ni, and Se. For example, copper is present in certain respiratory-chain components and enzymes.

9Cell Nutrients- Micronutrients

3 Trace elements rarely required are B, Al, Si, Cr, V, Sn, Be, F, Ti, Ga, Ge, Br, Zr, W, Li and I. These elements are required in concentrations of less than 10-6M and are toxic at high concentration.

4 Growth factor is also a micronutrient. Stimulatesthe growth and synthesis of some metabolites. e.g. vitamin, hormones and amino acids. They are required less than 10-6M.

Growth Media

There are two types of growth medium: defined medium and complex medium.

contains specific amounts of pure chemical compounds with known chemical compositions.

Glucose (30g/L), (NH4)2HPO4 (6g/L), NH4Cl (1.32 g/L), MgS04.7H2O (0.6 g/L), CaCl2 (0.05 g/L), KH2PO4 (10.0 g/L).

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Defined Media

- Results are more reproducible and the operator has better control of the fermentation.

- Recovery and purification processes are easier.

Growth Media

contains natural compounds whose chemical composition is not exactly known.- yeast extract, peptone, molasses or corn steep.- high yields: providing necessary growth factor.- cheaper than defined medium- more complex separation

Glucose (30g/L), yeast extract, NH4Cl (1.32 g/L), MgS04.7H2O (0.11 g/L), CaCl2.2H2O (0.08 g/L), K2HPO4 (2.0 g/L)for ethanol production by S. cerevisiae.

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Summary of Cell Nutrients

Nutrients required by cell living can be categorized into macronutrient that are required higher than 10-4M, micronutrients that less than 10-4M.

Macronutrients include N, C, O, H, S, P, K and Mg. They are major components in cell dry weight.

Micronutrients are classified into most widely needed elements, needed under specific conditions and rarely needed ones.

Growth medium can be either defined or complex.

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Stoichiometry of microbial growthand product formation

Microbial growth

Product formation

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