54 Seafoodchilting, refrigeration and freezing (Wiley Blackwell)

Science and Technology. Nalan Gkoglu and Pinar Yerlikaya

3.3.4 Enzymatic changes

The initial quality loss in fish is basically due to autolytic changes and is not related to
microbiological activity. The autolytic spoilage changes precede the other changes responsible
for the loss of fish quality during storage. Autolysis is principally enzymatic reactions, which take
place in fish tissues. Enzymes and other related chemical reactions do not inmediately cease
their activities in the fish muscle after fish death. Their continuation initiates other processes like
rigor-mortis; which is the basis for autolytic spoilage in fish {Huss f 995).

Enzymes are protein-like substances present in the flesh and stomach of fish and shellfish
which initiate or speed up chemical reactions. When the fish is alive, enzymes are usually kept
in balance with the help of digestive or blood systems. They remain active after the death of the
fish and are particularly involved in flavour changes that take place during the first few days of
storage before bacterial spoilage becomes significant a short time enzymatic activity can also
alter the texture and appearance of the flesh (Wheaton E Lawson l9B5). Autolysis is the
breakdown or decomposition of larger molecules such as proteins, lipids and carbohydrates
under the influence of enzymes up on the fish death. The quality of fish as a raw material for
consumption or for processing

depends largely on proteolysis, which is the autolysis of proteins (wakjira 20r l )' when captured
or harvested, fish and shellfish usually contain food in, their gut, and powerfuI enzymes are
present. upon the death of the animal. the enzymes penetrate the gut wall and surrounding
flesh, weakening and softening them. The gut and flesh may then, be invaded by spoilage
bacteria (Wheaton & Lawson t9S5). Enzymes play a role in the development of rigor mortis,
which is the progressive stiffening of muscles several hours after death. The stiffening effect is
as a result of coagulation of muscles protein. The duration of intensity of rigor depends upon the
species, temperature and condition of fish. it usually passes before bacteria invade the muscle,
leaving the flesh soft and limp. Following rigor thenself - digestion process commences, as a
result of enzymatic activity. The gut enzymes are particularly active at this time. A phenomenon
known as 'burst belly' ca' occur in just a few hours in some fish, such as sardine and herring,
and is caused by weakening of the belly wail due to self-digestion (Wheaton & Lawson t9S5).

The initial quality loss is mainly explained by autolytic changes, such as degradation of
,nucleotides (ATP-related compounds) by autolytic enzymes. The loss of the intermediate
'nucleotide, inosine monophosphate (IMP)), is responsible for the loss of fresh fish flavour.
These autolytic changes make catabolites available for bacterial growth (Huss 1995). Most
important in the autolytic spoilage changes is the degradation of fish nucleotides. Adenosine
triphosphate (ATP) degrades to adenosine diphosphate (ADP), adenosine monophosphate
(AMP), IMP, inosine (Ino) and hypoxanthine (Hx), associated with bitter fish flavours. generally,
the biochemical changes due to enzymatic activity related to freshness deterioration in fish are
change in the flavour and colour. The ultimate degradation of ATP intermediate nucleotides
(IMP and inosine) to Hx is attributed to bacterial activity (Haard 20a21). However, the presence
of ATP intermediate nucleotides, monophosphate (IMP) and inosine ([Ino) is associated with the
desirable sweet flavour in fresh fish (Huss 1995). The ATP of fish muscle breaks down, either
during the death struggle, or subsequently. This breakdown results in Iiberation of IMP, the
contributor of the pleasant flavour of fresh fish. The degradation of IMP to hypoxanthine is a
factor in the progressive loss of desirable flavour and in the development of bitter of-flavour
(Mandal & Mukherjee 1974)l. the first autolytic processes in the fish muscle tissue involve the
carbohydrates and the nucleotides. For a short period, the muscle cells continue the normal
physiological processes but soon the production of ATP stops. ATP functions as an ubiquitous
energy donor in
numerous metabolic processes. In the live organism, ATP is formed by the reaction between
ADP and creatine phosphate,, tire latter being a reservoir of energy-rich phosphate in tire
muscle cells. When the reservoir is depleted, the ATP is regenerated from ADP by
phosphorylation during glycolysis. After death, when the regeneration ceases, the ATP is rapidly
degraded. At low ATP levels rigor-mortis develops (Huss 1988). It is well known that enzymes
from the digestive tract play al important role in the autolysis of whole, uneviscerated fish-
During periods of heavy feeding the belly of certain fish is very susceptible to tissue degradation
and may burst within a few hours of catch (Huss l988). Production of lactic acid tends to lower
PH towards 6.4, depending on species, causing liberation and activation of tissue proteases.
Proteolysis may result in an increase in free amino acids but the rate seems to vary narkedly
with species. The most noticeable physical change resulting from autolysis in the ear§ stages of
deterioration is bursting of the bely walls of ungutted fish that have been eating heavily. The
concentration and activity digestive enzymes are highly in the guts and soon being to digest the
gut walls and surrounding tissues (Whittle et at. L99O\. Glycolysis is the anaerobic formation in
muscle of lactate from
glycogen by glycolysis, which continues for a time to provide energy to maintain muscle
functions. Glycolytic changes are rapid and the rate is considerable even at temperatures just
below 0"C. In general, fish muscles contain a relatively low amount of glycogen compared with
mammalian muscle and the final post-mortem pH is consequently higher; this makes fish meat
more susceptible to microbial attack.

However there are great variations in the glycogen content o{ different species; for example,
tuna have levels comparable with those found in mammals, and also within the same species.
As a rule, well-rested fish contain more glycogen than exhausted fish, well fed more than
starved fish and large more than small fish. Within the fish, glycogen is more concentrated in the
dark muscle than in the white muscle (Wakjira 2O11). Figure 3.3 shows glycogen breakdown
after death. Among postharvest changes, degradation of fish muscle caused try endogenous
proteases is a primary cause of quality losses during cold storage of handling (Haard L9941).

Proteases are able to hydrolyse the nuscle proteins. Microbial proteases may also be a potential
cause of proteolytic degradation. Proteolytic enzymes are found in all tissue, although both the
distribution of different enzymes and their activities show considerable variation. The highest
activities are found in fractions such as viscera and level but there are significant proteolytic
activities in muscle tissue as well, where the enzymes play an important role in protein turnover.
Alter death, the biological regulation of the enzymes is lost, and enzymes hydrolyze muscle
proteins and resolve the rigor-mortis corltraction (Hultruann 2003).