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Characterization of Pacific whiting protease and food-grade inhibitors for surimi productionWeerasinghe, Vasana C. 28 April 1995 (has links)
Cathepsin B was the most active cysteine proteinase in the Pacific whiting
(Merluccius productus) fish fillet, and cathepsin L in surimi when the activities of the
most active cysteine proteinases (cathepsin L, B, and H) were compared. Cathepsin L
showed maximum activity at 55°C in both fish fillet and surimi, indicating its function
in myosin degradation during conventional cooking of fish fillet and surimi. Washing
during surimi processing removed cathepsin B and H but not cathepsin L. Autolytic
analysis of surimi proteins showed that the myosin was the primary target, while actin
and myosin light chain showed limited hydrolysis during 2 hr incubation. When
purified Pacific whiting proteinase was incubated with various component of fish
muscle, proteinase was capable of hydrolyzing purified myofibrils myosin, and native
and heat-denatured collagen. The degradation pattern of myofibrils by the proteinase
was the same as the autolytic pattern of surimi.
Inhibition by the food-grade proteinase inhibitors varied with the catalytic type
of proteinase. Beef plasma protein (BPP) had a higher percentage of papain inhibitors, followed by whey protein concentrate (WPC), potato powder (PP), and egg white
(EW). On the other hand, EW had a higher percentage of trypsin inhibitors followed
by BPP, PP, and WPC. EW inhibited trypsin activity completely at levels as low as
1%. WPC inhibited the autolytic activity of fresh surimi. Bovine serum albumin
(BSA) was not effective as WPC. WPC can be used as an inhibitor for the Pacific
whiting surimi, but high concentration is required.
A limited number of inhibitory components were found, as the components in
food-grade inhibitors were characterized by inhibitory activity staining. Both EW and
PP showed more serine proteinase inhibitors than cysteine proteinase inhibitors. PP
showed one cysteine inhibitory component while EW did not show any. BSA in both
WPC and BPP acts as an nonspecific competitive inhibitor and reduces the enzyme
activity. An unidentified high molecular weight protein (HMP) found in WPC, BPP,
and BSA functions as an alternative substrate for papain while it functions as true
inhibitor for trypsin. / Graduation date: 1995
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Kinetic properties and characterization of purified proteases from Pacific whiting (Merluccius productus)Wu, JuWen 10 March 1994 (has links)
Kinetic properties of the two proteases, causing textural degradation of Pacific
whiting (Merluccius productus) during heating, were compared and characterized with the
synthetic substrate, Z-Phe-Arg-NMec. Pacific whiting P-I and P-II showed the highest
specificity on Z-Phe-Arg-NMec, specific substrate for cathepsin L. The K [subscript m] of
preactivated P-I and P-II were 62.98 and 76.02 (μM), and k [subscript cat], 2.38 and 1.34 (s⁻¹)
against Z-Phe-Arg-NMec at pH 7.0 and 30°C, respectively. Optimum pH stability for
preactivated P-I and P-II is between 4.5 and 5.5. Both enzymes showed similar pH-induced
preactivation profiles at 30°C. The maximal activity for both enzymes was
obtained by preactivating the enzyme at a range of pH 5.5 to 7.5. The highest activation
rate for both enzymes was determined at pH 7.5. At pH 5.5, the rate to reach the
maximal activity was the slowest, but the activity was stable up to 1 hr. P-I and P-II shared similar temperature profiles at pH 5.5 and pH 7.0 studied. Optimum temperatures
at pH 5.5 and 7.0 for both proteases on the same substrate were 55°C. Significant
thermal inactivation for both enzymes was shown at 75°C. Preactivated P-I and P-II
displayed a similar first order thermal inactivation profile at pH 7.0. At 30 and 90°C, half
lives, t [subscript 1/2], for Pacific whiting P-I were 49.50 and 0.20 min and for P-II, 32.54 and 0.18
min, respectively. The rate constant of inactivation for both proteases increased about
200-fold between two limits, 30 and 90°C. Half lives at 55°C, optimum temperature, for
P-I and P-II were also determined to be 5.29 and 6.75 min. The increase in thermal
inactivation rate constants independent of substrates corresponded to an activation energy
for heat denaturation of 21.18 kcal/mol for P-I and 19.97 kcal/mol for P-II by Arrhenius
plot. These similar kinetic properties, i.e., kinetic parameters, pH profile and thermal
inactivation rate constant, suggested that Pacific whiting P-I and P-II are the same
enzyme. / Graduation date: 1994
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Categorisation and chemical composition of Cape hake (Merluccius ssp.) wasteRoelf, Craig Ashley 03 1900 (has links)
Thesis (MScAgric)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: Cape hake (Merluccius capensis and M. paradoxus) is commercially the most
important trawl-caught fish off the South African, coastline and due to current
intensive fish processing procedures Cape hake contributes the most to the total fishwaste
production. Besides its commercial importance fish is also regarded as one of
the single most important consumable natural resources, either in the raw or frozen
form. Most of South Africa's commercially trawled demersal fish has already been
partially cleaned (i.e. headed and gutted) before landing with non-marketable bycatch
and hake-waste normally disposed of as discards, resulting in a waste of a potential
protein source.
This study was thus aimed at fulfilling several objectives namely: observing the
current large-scale commercial Cape hake harvesting procedure; constructing
prediction models for several morphological parameters (whole hake mass, headed &
gutted hake mass, hake head mass, hake head length, hake head breadth and hake
head height) of Cape hake (Merluccius ssp.), using whole hake length as the
independent variable; and determining the chemical composition (moisture, protein,
fat, ash, macro and trace elements) of several hake head sections (clean head, neck
flesh, tongue, tongue cartilage, jaw, gills, heart, intestines, gut, kidney, kidney &
kidney bone and gut & gall); determining the effect that storage has on the fatty acid
profile of both the clean head and neck flesh sections. The results obtained would
supply necessary data required for techno-economic investigations in the use of hake
heads.
For each of the six prediction models constructed, there was an increase in the
variance of the data points of categories 3 (64-80 cm) and 4 (>80 cm) as opposed to
categories 1 (30-46 cm) and 2 (47-63 cm). This could be attributed to a smaller
sample set for both categories 3 and 4 or due to an expected increase in the variance
when investigating larger biological samples. There was also a clustering of data in
the three areas for each prediction model namely, within category 1 and across
categories 2 and 3 and 3 and 4. This emphasised the latitudinal stratification of the
Cape hake population by age, hence their stratification by size. The prediction
models constructed for both boat trips 2 and 3 differed significantly (p<0.01) from that of boat trip 1, with the exception of the hake head length (cm) prediction model. The
constructed prediction models, for each of the three respective boat trips, showed
good predictive abilities as was indicated by the low Mean Square Error (MSE) values
for the test sets, and high Pearson's correlation coefficient (r) values. These
prediction models can be used in the fishing industry with confidence for Cape hake
within the time frame each respective boat trip was carried out.
The neck flesh could be regarded as the most important concerning chemical
composition whereas the jaw could be seen as the most important when one
considers mineral content. This therefore means that the jaw section, once
appropriately processed is a potential Ca, Na and Fe source for supplementing diets
of people suffering from a Ca, Na or Fe deficient diet. With regard to chemical status
the neck flesh section is seen as a good potential source of both protein and fat,
which could be attributed to the fact that hake muscle constitutes a major portion of
this section. This section could thus be used to supplement the protein and fat of an
existing food product, which is protein and fat deficient for people suffering from a
protein and fat deficient diet.
Similarly, a market could be created for the production of an economical food
product with the neck flesh section being the main ingredient. Once this have been
accomplished, fishing vessels may be persuaded to retain their Cape hake fish-waste
for further processing due to the value of the prepared food products and thereby
maintain profitability while abiding to governmental law.
In conclusion non-government scientists should have more input in the
decision-making process concerning matters affecting South Africa's marine
biodiversity in order for future key policy and legislation drafts to be effective.
Improvement of current fish preservation techniques and the known chemical
composition of currently discarded material will result in informed decisions of future
matters concerning its disposal. / AFRIKAANSE OPSOMMING: Kaapse stokvis (Merluccius capensis en M. paradoxus) is kommersieel Suid-Afrika se
belangrikste vis spesie. Aangesien die Suid-Afrikaanse visprosesseringsbedryf baie
intensief is, dra die Kaapse stokvis verwerkingsindustrie die grootste gedeelte by tot
die totale visafval produksie. Die meeste van Suid-Afrika se visvangste word
gedeeltelik skoongemaak voor landing terwyl nie-kommersiële byvangste en visafval
gewoonlik oorboord gegooi word tydens die vangproses. Dit lei tot die vermorsing
van 'n potensïele proteïen bron.
Hierdie studie was dus gemik om: die huidige grootskaalse kommersiële
Kaapse stokvis visvangsproses waar te neem; voorspellingsmodelle vir verskeie
morfologiese parameters (heel vis massa, vis massa sonder kop en binnedele,
stokvis kop massa, stokvis kop lengte, stokvis kop breedte en stokvis kop hoogte) vir
Kaapse stokvis (Merluccius ssp.) te ontwikkel deur die hele lengte van die vis te
gebruik as die onafhanklike veranderlike; die chemiese samestelling (vog, proteïen,
vet, as, makro en spoor elemente) van verskillende dele van die viskop (skoonkop,
nekweefsel, tong, tong kraakbeen, kaak, kiewe, hart, ingewand, derm, nier, nier &
nierbeen en derm & gal); sowel as die effek van opberging op die vetsuurprofiel van
beide die skoonkop en nekweefsel dele van die Kaapse stokvis kop. Hierdie resultate
sal dan gebruik word vir die tegnies-ekonomies ondersoek in die gebruik van Kaapse
stokvis koppe.
Vir elk van die ses voorspellingsmodelle ontwikkel, was daar 'n vermeerdering
in die variansie van die datapunte vir kategorieë 3 (64-80 cm) en 4 (>80 cm) teenoor
kategorieë 1 (30-46 cm) en 2 (47-63 cm). Dit kan moontlik wees as gevolg van die
kleiner monster trekking vir beide kategorieë 3 en 4 of as gevolg van verwagte
toename in variansie wanneer groter biologiese monsters ondersoek word. Daar was
ook 'n groepering van data in drie plekke vir elke voorspellingsmodel naamlik; binne
in kategorieë 1 en oor kategorieë 2 en 3 en 3 en 4. Dit beklemtoon die geografiese
breedte van die Kaapse stokvis populasie op grond van ouderdom, en dus die
geografiese breedte op grond van grootte. Die voorspellingsmodelle ontwikkel vir
beide die tweede en derde bootvangs het betekenisvol verskil (p<0.01) van die eerste
bootvangs, behalwe die vir die stokvis kop lengte (cm) voorspellingsmodel. Die voorspellingsmodelle vir elk van die bootvangste het goeie voorspellingsvermoë
getoon wat bewys is deur die lae Gemiddelde Kwadraat Fout waardes vir toetsgroepe
en hoë Pearson's korrelasie koeffisiënt (r) waardes. Hierdie voorspellingsmodelle wat
ontwikkel is, kan dus met vertroue in die Kaapse stokvis visvangsbedryf gebruik word
mits dit ooreenstem met die periode waarin elke bootvangs uitgevoer was.
Die nekweefsel gedeelte is die mees belangrikste met betrekking tot chemiese
samestelling en die kaak die belangrikste in terme van minerale samestelling van die
verskeie viskop dele. Die kaak is dus, as dit voldoende geprosesseer word, 'n goeie
potensïele bron van Ca, Na en Fe en kan dus gebruik word om die dieet van mense
wat 'n gebrek het aan hierdie minerale aan te vul. Met betrekking tot die chemiese
samestelling van die nekweefsel gedeelte kan dit beskou word as 'n goeie potensiële
bron van beide proteïen en vet, wat toegeskryf kan word aan die feit dat spierweefsel
'n groot deel uitmaak van hierdie viskop gedeelte. Hierdie viskop gedeelte sal dus
uitstekend wees om die proteïen- en vetinhoud van 'n voedselproduk wat van nature
'n lae proteïen- en vetinhoud het te verhoog en hierdie produk sou dan geteiken word
op daardie gedeelte van die gemeenskap wat 'n proteïen en vet tekort in hul dieet
het.
As dit eers alles in plek is, dan sal die visvangs bedryf hul Kaapse stokvis afval
behou vir verdere prosessering deurdat dit gebruik word om die voedingsinhoud van
bestaande voedsel soorte sal verbeter en terselfdertyd sal hulle aan wetgewing
voldoen. Gevolglik sal nie-regerings navorsers meer betrokke moet wees by die
besluitnemingsproses met betrekking tot sake wat die Suid-Afrikaanse mariene lewe
affekteer en wat toekomstige wetgewing meer effektief sal maak. Die verbetering van
huidige vis preserveringstegnieke gepaardgaande met die kennis van die chemiese
samestelling van die Kaapse stokvis koppe sal lei na beter toekomstige besluite oor
die afset daarvan.
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