Partial Purification and Characterization of a Thermostable Trypsin From Pyloric Caeca of Tambaqui (Colossoma Macropomum)

Bezerra, Ranilson S.; Santos, Juliana Ferreira dos; Paiva, Patrícia Maria Guedes; Correia, Maria T.S.; Vieira, Vera L. A.; Carvalho, Luiz Bezerra de

doi:10.1111/j.1745-4514.2001.tb00734.x

Cited by 71 publications

(86 citation statements)

References 22 publications

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“…Freitas-Júnior et al (2012) purified a trypsin extracted from giant Amazonian fish pirarucu (Arapaima gigas) viscera which showed an optimal temperature of 65 C. The purified trypsin was stable at temperature range 25e55 C for 30 min, losing only about 10% of its activity at 60 C (Table 2). Bezerra et al (2001) reported an optimal temperature of 65 C for trypsin extracted from tambaqui (Colossoma macropomum) and observed that the enzyme was stable at 55 C up to 30 min. In the same context, Bougatef et al (2007) purified a trypsin from sardine viscera which retained more than 80% of its initial activity after 4 h of incubation at 50 C.…”

Section: Temperaturementioning

confidence: 98%

Trypsins from fish processing waste: characteristics and biotechnological applications – comprehensive review

Bougatef

2013

Journal of Cleaner Production

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Section: Temperaturementioning

confidence: 98%

Trypsins from fish processing waste: characteristics and biotechnological applications – comprehensive review

Bougatef

2013

Journal of Cleaner Production

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“…It is also speculated that differences in the level of purification fold after ATPS process among tuna species might be related to different physicochemical and enzyme properties. Most of the methods reported for proteinase purification from fish digestive organs involved several steps, including ammonium sulfate precipitation, size-exclusion and ion-exchange chromatography [14,15], hydrophobic interaction chromatography [5] and affinity chromatography [29,38]. In view of their characteristics, these multi-step methods result in high cost and time consuming purification process.…”

Section: Recovery Of Spleen Proteinase From Other Tuna Speciesmentioning

confidence: 99%

“…Bezerra et al [14] partially purified trypsin from pyloric caeca of tambaqui (Colossoma macropomum) and found that the enzyme had an optimal pH of 9.5. Byun et al [15] purified and characterized serine proteinases from pyloric caeca of tuna (Thunnus thynnus).…”

mentioning

confidence: 99%

Partitioning and recovery of proteinase from tuna spleen by aqueous two-phase systems

Klomklao

Benjakul

Visessanguan

et al. 2005

Process Biochemistry

101

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Partitioning of spleen proteinase from yellowfin tuna (Thunnus albacores) in an aqueous two-phase system (ATPS) was investigated. Phase compositions including PEG molecular mass and concentration as well as types and concentration of salts affected the protein partitioning. ATPS comprising PEG1000 (15% w/w) and magnesium sulfate (20% w/w) provided the best condition for the maximum partitioning of the proteinase into the top phase and gave a highest specific activity (47.0 units/µg protein) and purification fold (6.61). The yield of 69.0% was obtained.Under the same ATPS condition used, the partitioning of proteinase of splenic extract from three tuna species involving skipjack tuna, yellowfin tuna and tongol tuna were compared. The purity of splenic extract from all tuna species increased after ATPS process. Among all species tested, yellowfin tuna showed the highest purification fold, followed by tongol tuna and skipjack tuna, respectively. SDS-substrate gel electrophoresis revealed that the band intensity of major proteinase in ATPS fraction from all tuna species slightly increased with the concomitant decrease in band intensity of other contaminating proteins, indicating the greater specific activity of splenic extract. Therefore, ATPS was an effective method for partitioning and recovery of proteinases from tuna spleen.

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“…This step resulted in an increase in the specific activity. Heat treatment has proven to be an important strategy for purification of thermostable enzymes; it denaturates thermolabile proteins in the crude extract (Bezerra et al 2001). Then, the heated crude enzyme was fractionated with ammonium sulphate.…”

Section: Sod Purificationmentioning

confidence: 99%

A heat-stable Cu/Zn superoxide dismutase from the viscera of sardinelle (Sardinella aurita): purification and biochemical characterization

et al. 2014

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Superoxide dismutase (SOD) is an enzyme that protects against oxidative stress from superoxide radicals in living cells. This enzyme was extracted from sardinelle (Sardinella aurita) viscera, purified and characterized. The Cu/Zn-SOD was purified to homogeneity by the three-step procedure consisting of the heating at 65• C for 15 min, precipitation with ammonium sulphate (30-60%, w/v) and Sephadex G-100 gel filtration with a 7.17-fold increase in specific activity. The molecular weight of the native enzyme was estimated to be 40 kDa by G-125 gel filtration on HPLC column and that of the subunit mass, deduced by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, was 20 kDa. Thus the native enzyme appeared to be a homodimer. The optimum pH and temperature for the purified SOD activity were determined to be pH 7.0 and 40• C, respectively. It retained more than 85% of its initial activity after 1 h of incubation at 50• C. The enzyme had a broad stability pH range of 6.0-9.0. The N-terminal sequence of the purified enzyme was VLKAVCVLKGTGEVT. This sequence exhibited a high degree of sequence similarity with other fish Cu/Zn SODs.

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Partial Purification and Characterization of a Thermostable Trypsin From Pyloric Caeca of Tambaqui (Colossoma Macropomum)

Cited by 71 publications

References 22 publications

Trypsins from fish processing waste: characteristics and biotechnological applications – comprehensive review

Trypsins from fish processing waste: characteristics and biotechnological applications – comprehensive review

Partitioning and recovery of proteinase from tuna spleen by aqueous two-phase systems

A heat-stable Cu/Zn superoxide dismutase from the viscera of sardinelle (Sardinella aurita): purification and biochemical characterization

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