Partial purification and characterization of trimethylamine-N-oxide demethylase from lizardfish kidney

Trimethylamine-N-oxide demethylase (TMAOase) from lizardfish (Saurida micropectoralis) was partially purified by acidification and diethylaminoethyl (DEAE)-cellulose chromatography. The enzyme was purified 82-fold with a yield of 65.4%. The optimum pH and temperature were 7.0 and 50 degrees C, respectively. TMAOase was stable to heat treatment up to 50 degrees C and the activation energy was calculated to be 30.5 kJ mol(-1) K(-1). Combined cofactors (FeCl(2), ascorbate and cysteine) were required for full activation. FeCl(2) exhibited a higher stimulating effect on TMAOase activity than FeCl(3). At concentration less than 2 mM, ascorbate was more stimulatory to the activity than cysteine. The activity was tolerant of NaCl concentration up to 0.5 M. The enzyme had a K(m) for TMAO of 16.2 mM and V(max) of 0.35 micromol min(-1) and was able to convert TMAO to dimethylamine (DMA) and formaldehyde. The molecular mass of enzyme was estimated to be 128 kDa based on activity staining.     

Combining Effect of Microbial Transglutaminase and Bambara Groundnut Protein Isolate on Gel Properties of Surimi from Sardine (Sardinella albella)

Effects of protein isolate from bambara groundnut (BGPI) at different levels (0–6 %, w/w) in combination with microbial transglutaminase (MTGase) at different concentrations (0, 0.3 and 0.6 U g^?1surimi) on gels properties of sardine (Sardinella albella) surimi were investigated. In the absence of MTGase, the increases in breaking force and deformation of gels were obtained when BGPI at levels of 1.5–3 % was incorporated (P?<?0.05). The further increases in BGPI levels (4.5–6 %) resulted in the decrease in breaking force and deformation (P?<?0.05). When MTGase (0.3 and 0.6 U g^?1surimi) was added, the increase in breaking force and deformation were noticed, regardless of BGPI levels, and the strengthening effect was in dose-dependent manner. The increases in hardness, gumminess and chewiness were also observed when surimi gel was added with BGPI and MTGase (P?<?0.05). Water-holding capacity of gels was improved with increasing level of BGPI, and MTGase incorporated (P?<?0.05). Whiteness of gels slightly decreased with increasing BGPI levels, however the addition of MTGase had no impact on whiteness (P?>?0.05). Based on electrophoretic study, myosin heavy chain decreased with addition of MTGase, indicating the formation of cross-links. More compact structure was observed in gel added with MTGase (0.6 U g^?1surimi) and 6 % BGPI, and was accompanied by an increased gel strength.     

Identification and histamine formation of Tetragenococcus isolated from Thai fermented food products

Forty-one tetrad-forming halophilic lactic acid bacteria were isolated from 7 kinds of fermented foods in Thailand. All the isolates were identified as the genus Tetragenococcus by their phenotypic characteristics. On the basis of 16S rRNA gene restriction analysis using MboI and AluI and 16S rRNA gene sequence analyses, 41 isolates could be divided into two groups (groups A and B). All 22 isolates in Group A were identified as T. halophilus. 16S rRNA gene sequences of the representative isolates, SP37-2 and KS87-1 exhibited 99.4–99.5 % similarity to that of T. halophilus ATCC 33315^T. Nineteen isolates in Group B were identified as T. muriaticus. 16S rRNA gene sequences of the representative isolates, KM1-5 and KS87-14, showed 99.0–99.6 % similarity to that of T. muriaticus JCM 10006^T. Histamine formation was determined by using HPLC and the histidine decarboxylase (hdc) gene of the newly isolated histamine-producing strain was partially sequenced. The strain KS87-14 prolifically formed histamine 10 times higher than the reported T. muriaticus JCM 10006^T. The positive detection of KS87-14 was achieved by using hdcA gene-specific primers JV16HC and JV17HC.     

Characteristics of film based on protein isolate from red tilapia muscle with negligible yellow discoloration

The properties of film from fish protein isolate (FPI) prepared by prior washing followed by alkaline solubilization process (ASP) from red tilapia muscle were monitored during the storage of 20 days at 50% RH and 25°C, in comparison with those of films from washed mince. Lipid, heme iron and non-heme iron contents in FPI were decreased by 98.8, 36.8 and 91.9%, respectively in comparison with those of washed mince (p<0.05). Films from FPI had higher tensile strength (TS) and elongation at break (EAB) than those from washed mince for both pH (3 and 11) used for film preparation (p<0.05). Film from FPI at pH 3 showed the highest TS, while that from washed mince at pH 11 had the lowest TS (p<0.05). Nevertheless, films from FPI had higher WVP than those from washed mince for both pH used (p<0.05). At the same pH used for film preparation (3 or 11), films from FPI showed the lower TBARS values than those from washed mince (p<0.05). Nevertheless, films from both FPI and washed mince had the higher TBARS values when pH 3 was used for film preparation, compared with pH 11 (p<0.05). Among all films, those from FPI prepared at pH 3 had the highest transparency and no yellow discoloration was observed during the storage of 20 days, in comparison with other films (p<0.05). Conversely, film from washed mince prepared at pH 3 had the higher increase in b*-value and ΔE*-value than other films. Therefore, FPI could serve as a potential material for film preparation with lower contents of lipid and prooxidants, thereby preventing the yellow discoloration of the fish myofibrillar protein-based film during extended storage.     

Effect of Melanin-Free Ink on Mechanical Properties and Yellow Discolouration of Protein Film from Washed Sardine Mince

The effect of the melanin-free ink (MFI) at different concentrations (0, 0.1, 0.2, 0.3 and 0.4 gL^?1 of film forming solution (FFS)) on the properties and yellow discolouration of films from washed sardine mince was studied. Tensile strength (TS) of the film increased with increasing MFI concentration (P < 0.05). Conversely water vapour permeability (WVP) of films decreased as the concentration of MFI increased (P < 0.05). Fourier transform infrared (FTIR) spectra of films showed a slight shift to lower wavenumber of amide-B band of film added with MFI, indicating protein cross-linking. The microstructure showed slightly rough surface of the films when amount of MFI increased. However, cracks in the films were much decreased when MFI at higher levels was incorporated. Films added with MFI had lower thiobarbituric acid reactive substances (TBARS) value, indicating the lowered lipid oxidation. The addition of MFI also decreased the yellow discolouration but increased the transparency of film in a dose-dependent manner.     

The influence of storage conditions of tuna viscera before fermentation on the chemical, physical and microbiological changes in fish sauce during fermentation

Effect of storage condition of tuna viscera on chemical, physical and microbiological changes of its sauce were monitored. Results based on microbial counts, protein degradation products, total volatile base (TVB), and trimethylamine (TMA) contents, showed that tuna viscera stored at room temperature underwent more deterioration than that kept in ice, especially with increasing storage time. As a result, fish sauce obtained from tuna viscera stored at room temperature for a longer time rendered the greater amino nitrogen, TVB, TMA contents as well as browning intensity. However, storage conditions had no marked effect on overall changes in chemical, physical and microbiological characteristics of sauce generated during fermentation. Additionally, fish sauce produced from tuna viscera kept at room temperature comprised lower histamine content than that prepared from fresh or ice-stored viscera. Therefore, tuna viscera stored at room temperature for up to 8h could be used for the production of fish sauce with no detrimental effect on the quality.     

Purification and characterization of two pepsins from the stomach of pectoral rattail (Coryphaenoides pectoralis)

Two pepsins (A and B) were purified from the stomach of pectoral rattail (Coryphaenoides pectoralis) by acidification, ammonium sulfate precipitation, gel filtration chromatography and anion exchange chromatography to obtain a single band on native-PAGE and SDS-PAGE. The purities of pepsin A and B were increased to 7.1- and 13.0-fold with approximately 5.7% and 2.2% yield, respectively. Pepsin A and B had the apparent molecular weights of 35 and 31 kDa, respectively, when analyzed using SDS-PAGE and Sephacryl S-200 gel filtration. Pepsin A and B showed maximal activity at pH 3.0 and 3.5, respectively, and had the same optimal temperature at 45 °C using hemoglobin as a substrate. Both pepsin A and B were stable in the pH range of 2.0–6.0 but were unstable at the temperatures greater than 40 °C. Activity of both pepsins was inhibited by pepstatin A and was activated by divalent cations, indicating pepsin characteristics. Activities of both pepsins continuously decreased as NaCl concentration increased (0–30%). The enzymes had high affinity and activity toward hemoglobin with K_m and K_cat values of 98–152 μM and 32–50 S^− 1, respectively. Purified pepsins generally showed the similar characteristics to other fish pepsins.     

Physical and Textural Properties of Mayonnaise Prepared Using Virgin Coconut Oil/Fish Oil Blend

Physical and textural properties of mayonnaise prepared using virgin coconut oil (VCO)/fish oil (FO) blends at different ratios were examined in comparison with that prepared using soybean oil (SO) as affected by storage time (30 days). At day 0, sample prepared with SO showed the highest L*, a*, and b* values among all the samples, whereas the lowest values were noticeable for VCO containing sample. At day 30 of storage, decreases in L*, and b* values of all mayonnaise samples were observed (p < 0.05). However, a* values were increased at day 30 of storage (p < 0.05). For texture analysis, highest firmness, consistency and cohesiveness were obtained for the sample containing SO. Increasing levels of FO in VCO/FO samples increased the firmness, consistency and cohesiveness. For all the samples, loss modulus (G″) values were lower than G′. After 30 days of storage, all samples demonstrated slight decreases in G′ and viscosity than freshly prepared mayonnaise (day 0). When the sample containing VCO/FO (90:10) blend was further characterized, slight difference was observed in microscopic structure and droplet size distribution before and after storage of 30 days. Increase in droplet size was noticeable because of coalescence after the storage. Overall, type of oil used for preparation of mayonnaise as well as storage time affected the physical properties including textural and rheological properties of mayonnaise.

     

Surface activity and molecular characteristics of cuttlefish skin gelatin modified by oxidized linoleic acid

Surface activity and molecular changes of cuttlefish skin gelatin modified with oxidized linoleic acid (OLA) prepared at 60, 70 and 80 °C at different times were investigated. Modification of gelatin with OLA could improve surface activity of resulting gelatin as evidenced by the decreased surface tension and the increased foaming and emulsifying properties. Interaction between OLA and gelatin led to the generation of carbonyl groups, loss of free amino content and the increase in particle size of resulting gelatin. Emulsion stabilized by modified gelatin had the smaller mean particle diameter with higher stability, compared with that stabilized by gelatin without modification.     

Properties of film from splendid squid (Loligo formosana) skin gelatin with various extraction temperatures

Properties of film from splendid squid (Loligo formosana) skin gelatin extracted at different temperatures (50-80°C) were investigated. Tensile strength (TS) and elongation at break (EAB) of films decreased, but water vapour permeability (WVP) increased (P<0.05) as the extraction temperature increased. Increase in transparency value with coincidental decrease in lightness was observed with increasing extraction temperatures. Electrophoretic study revealed that degradation of gelatin became more pronounced with increasing extraction temperatures. As a consequence, their corresponding films had the lower mechanical properties. FTIR spectra of obtained gelatin films revealed the significant loss of molecular order of the triple helix. Thermogravimetric analysis indicated that F80 exhibited the higher heat susceptibility and weight loss. Loosen structure was observed in film prepared from gelatin with increasing extraction temperatures. Thus, the temperature used for gelatin extraction from splendid squid skin directly affected the properties of corresponding films.