Chemical and Food Engineering Department

Republic of the Philippines
BATANGAS STATE UNIVERSITY

Pablo Borbon Main II, Alangilan, Batangas City
College of Engineering, Architecture & Fine Arts
www.batstate-u.edu.ph Tel. No. (043) 425-0139 loc. 118
Chemical and Food Engineering Department
ChE 526 BIOCHEMICAL ENGINEERING
NARRATIVE REPORT
Product Formation Kinetics
Submitted by:
John Bryan A. Aldovino
ChE-5202 (Sit-in)
Submitted to:
Engr. Mary Rose F. Persincula
Instructor
May 2020
Objectives:
 To discuss the several equations involving in the computational work for product
formation kinetics
 To differentiate and identify several kinetic models used for various situations
Introduction
The types of kinetic description that may be employed for product formation by cell
populations parallel those used to describe cell population growth. As we shall see in further
discussion, it is possible to formulate useful models of protein synthesis kinetics at the molecular
level, taking advantage of contemporary understanding of molecular controls
Unsturctured Models
The simplest types of product formation kinetics arise when there is a simple
stoichiometric connection between product formation and substrate utilization or cell growth.
Then, the product formation rate rfp may be written as:
r fp=Y P / S r fs
r fp=Y P / X r fx
The alcohol fermentation shown in batch culture in the following figure is an example of
this class – type I fermentations. Such product formation kinetics are sometimes called growth
associated.this class – type I fermentations. Such product formation kinetics are sometimes
called growth associated.
In many fermentations, especially those involving secondary metabolites, significant

product formation does not occur until relatively late in a batch cultivation, perhaps approaching
or into stationary phase. The penicillin fermentation data exemplify such behavior.
The Leudeking-Piret model
The now classic study of Leudeking and Piret on the lactic acid fermentation by
Lactobacillus delbruekii indicated product formation kinetics which combined growth-associated
and nongrowth-associated contributions:
The study considered the relationship of cell growth to product formation. When the cells
or some constituent of cells that is proportional to cell mass is the product, the rate of formation
of product directly relates to the rate of growth. Many other products are known that have zero
on a small rate of formation until growth ceases. It is useful to think of this as the cell using
resources to grow until some important nutrient such as the sugar falls to a low concentration.
This two-parameter kinetic expression, often termed Leudeking-Piret kinetics, has proved
extremely useful and versatile in fitting product formation data from much different
fermentation. This is an expected kinetic form when the product is the result of energy-yielding
metabolism, as in several anaerobic fermentation.
Table 7 E1.1 lists the kinetic parameter values determined by this procedure for four different
fermentations producing extracellular polysaccharides.
But when tested with the microbial process for producing lactic acid, this equation fit
actual data very well. This was somewhat fortuitous because the agreement is not so good with
some other processes. Probably alpha and beta do not remain constant as the physiological age of
the cells or the conditions of the process change. Nevertheless, the concept of distinguishing the
relationship of product formation to growth rate is sound.
The time course of product concentration in the medium during a batch fermentation can
be quite complicated, and more than one product may be formed and interconverted. F. H.
Deindoerfer introduced a classification of many of these possibilities which serves to illustrate
some possible cases. In some situations, complicated product formation kinetics may result from
changes in cell metabolic operation during the reaction.
In this section, we consider an interesting secondary metabolite production process which

serves as a useful introduction to several important aspects of many antibiotic fermentations as
well as to new considerations in kinetic modeling.
Synthesis of several microbial antibiotics and other secondary metabolites is inhibited by

high concentrations of phosphate.
Chemically Structured Product Formation Kinetics Models
In this model, the specific growth rate is described by the Teissier equation based on
intracellular phosphate concentration pi (g KH2PO4/g biomass; an intrinsic cell composition
variable. All other composition variables in this model have units of grams per liter culture). The
term k2x2 is used to describe the rate of cell lysis.
Gene transcription models aim at quantifying gene transcription based on knowledge of

the promoter function. Among the best-studied promoters is the so-called lac-promoter of E. coli,
which takes part in regulating expression of genes that are involved in lactose uptake, i.e., lactose
permease and ~-galactosidase.
Understanding the regulation of this promoter represented a major breakthrough in

molecular biology, and the history of how the mechanisms were unraveled is an excellent
introduction to modern molecular biology (Muller-Hill, 1996). The lac-promoter is also of
substantial industrial interest as it is often used to drive expression of heterologous genes
encoding recombinant proteins in E. coli.
The three genes coding for enzymes necessary for lactose metabolism in E. coli are
coordinated in a so-called operon, and gene expression is coordinately controlled by two
regulatory sites positioned upstream of the genes (see Fig. 7.16):
✘ Control at the operator by a repressor protein.
✘ Carbon catabolite repression at the promotor.
The repressor protein X; has two binding sites - one site that specifically ensures binding to
the operator (Xo) and one site which may bind lactose (S,.J. When lactose binds to the repressor
protein, its conformation changes so that its affinity for binding to the operator is significantly
reduced. Thus lactose prevents the repressor protein from binding to the operator, and
transcription of the genes by RNA polymerase is therefore allowed.
Consequently, lactose serves as an inducer of transcription; i.e., expression of the three genes
lacZ, lacY, and lacA is not possible unless lactose (or another inducer, e.g. isopropyl-~-D-
thiogalactoside, abbreviated IPTG) is present.' The binding of the repressor protein to lactose and
the operator may be described by reactions (7.44), where n = 4 is the number ofbinding sites for
lactose on the repressor protein.
Figure 7.16 The lac-operon of E. coli. The operon includes a gene (lac!) for the
repressor protein (r), promotor (P), operator (0), and the three genes lacZ, lacY, and lacA,
which code for different enzymes: lacZ codes for P-galactosidase, lacY for lactose
permease, and lacA for thio galactoside transacylase. In its free form the repressor protein
may bind to the operator; when it complexes with the inducer (i), conformational changes
of the repressor protein prevent binding to the operator. cAMP complexes with CAP and
the complex may bind to the promoter, whereby the RNA polymerase may start the
transcription from the promotor.
The model in Eq. (7.44) gives a simplified description of the true system since there may be
different binding affmities for the repressor protein depending on how much lactose is bound to
the protein (see Problem 7.3). With the concentration of the species (indicated with squared
brackets) being in moles per gram dry weight, the equilibrium constants Kj, i = 1,2,3,4 are given
by:
The quantity Q2 of Eq. (7.58) is used to model the repression effect of glucose, just as QI
in Eq. (7.52) is used to describe the induction of lactose on gene expression and hereby synthesis
of enzymes necessary for lactose metabolism. However, in order to apply Eq. (7.58), one needs
to know the intracellular level of CAP (which in a simple model may be assumed to be constant)
and also the level of cAMP. Harder and Roels (1982) suggest the following empirical correlation
between XcAMP and the extracellular glucose concentration Sglc:
Conclusion
Products are categorised into various types based on their relation with microbial growth.
There are three types of products. The first is Growth associated products then followed by Non-
Growth associated products and Mixed mode product formation. In growth associated products,
product is formed along with the growth of the microbial cells and product concentration is
almost directly proportional to microbial growth rate.
Whereas in non-growth associated products, product formation is nowhere related with

growth rate of microbial cells but its a function of cell concentration and we can say that product
formation and microbial growth rate are almost inversely proportional to each other. But with
third category product formation is a combination of both microbial cell concentration and
microbial growth rate.
References:
Baileys, J.E. and Ollis, D.F. (1986) Biochemical Engineering Fundamentals, 2nd edition.
McGraw-Hill Inc. Singapore.
Jagriti Singh, Nirmala Kaushik & Soumitra Biswas 2014. Bioreactors – Technology &
DesignAnalysis. The Scitech Journal: Vol 01 (6):28-36

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