Introduction

Internal cell functions associated with cell growth are not easily measurable; however, they
play important roles in kinetic behaviors. In this topic, the kinetic implication of endogenous
and maintenance metabolism, it will tackle about simplifying the cell growth-associated
functions with a focus on their effects on growth needs.
To harness the free energy produced by catabolic processes in terms of high-energy phosphate
bonds (in particular, in the form of ATP) for subsequent use in the biosynthesis of biomass
constituents in the anabolism, the cellular content of ATP (and ADP) must be controlled quite
rigorously. As the turnover time of ATP is low (Theobald et al., 1996), there must be a tight
balancing of the energy-forming reactions (catabolism) and the energy-utilizing reactions (the
anabolism) inside the cell. In an analogy to the tight balancing of synthesis and consumption
of ATP, the cell needs to balance the synthesis and consumption of the cofactors NADH or
NADPH, which have a small turnover time. Consequently the cell must exercise a strict control
of the level of these compounds as well.
The balances for ATP, NADH, and NADPH are used to relate the fluxes through different parts
of the metabolic network. However, in order to apply the ATP balance it is important that all
ATP forming and consuming reactions are considered; one group of energy consuming
processes inside the cell, namely the maintenance processes, is especially important, as it is
seemingly a drain on net energy without an apparent benefit to the system.
Cell Maintenance and Endogenous Metabolism
Cell maintenance is the consumption of a key substrate under consideration to maintain the
desired functionality of the cell in addition to the stoichiometrically quantifiable desired
outcome, such as cell growth. The balance of ATP, NADH, and NADPH, for example, requires
maintenance energy. Endogenous metabolism, on the other hand, refers to the metabolic needs
of a cell to stay viable either by consuming nutrients in the medium when available or
converting intracellular materials when starved. Therefore, strictly speaking, cell maintenance
is cell growth dependent, while endogenous metabolic needs are often growth independent.
Cell maintenance and endogenous metabolic needs are, however, closely related and not easily
separated in practice. Therefore, these two are often combined in bioprocess modeling, and
both terminologies are used interchangeably at times.
Herbert (1959) showed that it is necessary to consider the “endogenous metabolism,” or cell
death, when the substrate utilization for biomass growth is to be calculated. He assumed that
this endogenous metabolism results in a decrease of the amount of biomass, and he described
the degradation as a first-order process with a specific rate of biomass degradation μe.
Restitution of the degraded biomass requires substrate and the total substrate consumption is,
therefore
-rs = (YFS/X μG + YFS/X μe)X
Pirt (1965) introduced an empirical correlation identical in form to previous equation but he
collected the product of YFS/x and μe in the empirical constant mS:
-rs = (YFS/X μG + mS)X
This empirical constant was called the maintenance coefficient. Strictly speaking, maintenance
requirements and endogenous needs are different, and we tend to simplify the metabolic system
by “equate” the maintenance and endogenous needs. This allows us to lump the two different
indirect growth needs to one quantity. The maintenance coefficient can then be related to the
specific rate of substrate uptake for cellular maintenance, via:
−𝑟𝑠 | 𝑚𝑎𝑖𝑛𝑡𝑒𝑛𝑎𝑛𝑐𝑒
𝑚𝑠 = = 𝑌𝐹𝑆/𝑋 𝑘𝑑
𝑋
Maintenance coefficient (mS) is an important parameter in the fermentation process and has
great influence on the biomass and product yield.
Bauchop and Esden (1960) introduced the concept of ATP requirements for biomass synthesis
via the yield factor YFATP/X and proposed a balance equation that is analogous to equation
introduced by Herbert (1959).
-rs = YFATP/X μG + mATP

Here, rATP specifies the total formation rate of ATP in catabolic pathways (different from the
net formation rate of ATP, which is implicitly assumed to be zero in the equation). From precise
measurements of the metabolic products of the anaerobic metabolism, it is possible to calculate
the specific formation rate of ATP, that is, rATP. This may be used to find experimental values
for YFATP/X and mATP, as shown in many studies.

The Monod equation is an approximated growth rate from the complicated metabolic pathways.
The same approximation can be applied based on a key intermediate (eg, ATP), rather than the
substrate from the environment. The specific cell growth rate can then be approximated by:
𝑟𝑋 μ𝑚𝑎𝑥 𝑌
μ𝐺 = =
𝑋 𝐾𝑌𝐺 + 𝑌
The maintenance cost of the intermediate Y can be assumed to be in a fashion similar to any
other bioreactions. In other words, the maintenance (or endogenous) cost can be approximated
by:
𝑟𝑒 μ𝑒𝑚𝑎𝑥 𝑌
μ𝑒 = =
𝑋 𝐾𝑒𝑌 + 𝑌
Where the saturation constant KeY needs not to be identical to the saturation constant KY in the
cell growth rate. KeY may not be exactly zero either (for which a constant maintenance
coefficient prevails). There are two ways to express the cell maintenance cost: either to the cell
growth rate or to the substrate availability. It is not difficult to see that:
𝑟𝑒 μ𝑒𝑚𝑎𝑥 𝑆
μ𝑒 = =
𝑋 𝐾𝑒𝑆 + 𝑆
Since cell maintenance goes head to tail with cell growth, substituting the first two equation:
μ𝑒𝑚𝑎𝑥 𝐾𝑌𝐺μ𝐺
μ𝑒 =
𝐾𝑒𝑌μ𝑚𝑎𝑥 + (𝐾𝑌𝐺 − 𝐾𝑒𝑌 )μ𝐺

The equation for cell maintenance can be written as:

μ𝑒μ𝑚𝑎𝑥 μ𝐺
μ𝑒 =
𝐾𝑒μ + μ𝐺

Apoptosis
Apoptosis is the process of programmed cell death (PCD) that may occur in multicellular
organisms. PCD involves a series of biochemical events that lead to a variety of morphological
changes, including blebbing changes to the cell membrane, such as loss of membrane
asymmetry and attachment, cell shrinkage, nuclear fragmentation, chromatin condensation,
and chromosomal DNA fragmentation. Processes of disposal of cellular debris whose results
do not damage the organism differentiate apoptosis from necrosis.
In contrast to necrosis, which is a form of traumatic cell death that results from acute cellular
injury, apoptosis confers advantages during an organism’s life cycle in general. For example,
the differentiation of fingers and toes in a developing human embryo occurs because cells
between the fingers apoptose; the result is that the digits are separate. Between 50 and 70 billion
cells die each day due to apoptosis in the average human adult. For an average child between
the ages of 8 and 14, approximately 20 billion to 30 billion cells die a day. In a year, this
amounts to the proliferation and subsequent destruction of amass of cells equal to one’s body
weight.
Research on apoptosis has increased substantially since the early 1990s. In addition to its
importance as a biological phenomenon, defective apoptotic processes have been implicated in
an extensive variety of diseases. Excessive apoptosis causes hypotrophy, such as in ischemic
damage, whereas an insufficient amount results in uncontrolled cell proliferation, such as
cancer.
Apoptosis occurs when a cell is damaged beyond repair, is infected with a virus, or has
experienced stressful conditions such as starvation. Damage to DNA from ionizing radiation
or toxic chemicals can also induce apoptosis via the actions of the tumor-suppressing gene.
Apoptosis also plays a role in preventing cancer. If a cell is unable to undergo apoptosis because
of mutation or biochemical inhibition, it continues to divide and develop into a tumor. For
example, infection by papilloma viruses causes a viral gene to interfere with the cell’s protein,
an important member of the apoptotic pathway. This interference in the apoptotic capability of
the cell plays a role in the development of cervical cancer.
Before the actual process of cell death is precipitated by enzymes, apoptotic signals must cause
regulatory proteins to initiate the apoptosis pathway. This step allows apoptotic signals to cause
cell death or stop the process should the cell no longer need to die. Several proteins are
involved, but two main methods of regulation have been identified: targeting mitochondria
functionality or directly transducing the signal via adaptor proteins to the apoptotic
mechanisms. Another extrinsic pathway for initiation identified in several toxin studies is an
increase in calcium concentration within a cell caused by drug activity, which also can cause
apoptosis via a calcium-binding protease calpain.
PCD in plants has a number of molecular similarities to animal apoptosis, but it also has
differences, notably the presence of a cell wall and the lack of an immune system, which
removes the pieces of the dead cell. Instead of an immune response, the dying cell synthesizes
substances to break itself down and places them in a vacuole, which ruptures as the cell dies.
One can infer that cell death in multicellular organisms is a means to maintain the overall health
of the organism. The energy spent may be termed endogenous metabolism. This is a
magnification of endogenous metabolism in single cells.
 University of Mindanao, Davao City
Engineering Department

In Partial Fulfillment of the Requirement in

Biochemical Engineering
ChE 544

Kinetic Implication of Endogenous and Maintenance Metabolism

Submitted by:
John M. Gohiling

Submitted to:
Engr. Arjan C. Lingaya
Instructor

July 4, 2017