Characterization of Collagen from Sakhalin Taimen Skin as Useful Biomass.

RESEARCH BACKGROUND: Animal collagen has been widely utilized in foods, cosmetics and biomedical fields. The non-edible portion, such as fish skin and bones, are obtained during cooking. Most of them are currently discarded as wastes, although the nutritional value of the skin and bones is high. The non-edible portion needs to be reused in order to reduce environmental impact, as it is one of the sources of environmental pollution. EXPERIMENTAL APPROACH: Collagen was prepared by cold acetone treatment from Sakhalin taimen skin as a waste produced during cooking. Next, the colour, SDS-polyacrylamide gel electrophoresis, ultraviolet absorption, subunit composition, amino acid composition, denaturation temperature and attenuated total reflectance-Fourier transform infrared spectroscopy analyses were conducted to explore the properties of the collagen. Lastly, we attempted to improve the functional properties of the collagen for future applications using chemical modification technique (succinylation). RESULTS AND CONCLUSIONS: Cold acetone treatment easily removed the fats and pigments from the skin. The odourless and pure white collagen was obtained with high yield. The α3 chain did not exist in the collagen. Sakhalin taimen skin collagen had rich α-helix and low ß-sheet structures. Succinylation caused the secondary structural changes of the collagen molecule. Moreover, it made it possible not only to increase the viscosity of the collagen solution but also to improve the solubility of the collagen under the physiological conditions around pH=6. NOVELTY AND SCIENTIFIC CONTRIBUTION: This finding is the first report on the absence of the α3 chain from salmonid fish skin collagens. The succinylated collagen from Sakhalin taimen skins as useful biomass has potential to utilize in foods, cosmetics and related industries.

Characterization of collagen from emu (Dromaius novaehollandiae) skins.

Collagen from emu skins as a by-product was prepared. The skins were hardly solubilized in acetic acid, however were successfully solubilized on digestion with 10 % pepsin (w/w) for 4 days. The yield of pepsin-solubilized collagen (PSC) was about 27.3 %, on a raw weight basis. By SDS-PAGE and CM-Toyopearl 650 M column chromatography, the presence of a fourth subunit that was previously designated α4 was confirmed. The denaturation temperature of the PSC was 31.5 °C, about 6-7 °C lower than that from the porcine skins. ATR-FTIR analysis indicated that the helical arrangements of the PSC from emu skins existed and its structures of PSC were changed slightly due to the loss of N- and C-terminus domains in similar to that from the porcine skins. That is, the PSC from emu skins did not possess telopeptide chains as major portion of antigenic sites in collagen. The present study indicates that a large quantity of emu skins as by-products have potential as a good alternative source of high-quality collagen for industrial purposes in the foods, cosmetics, and pharmaceutical and biomedical fields.

Functional properties of autolysate and enzymatic hydrolysates from yam tsukuneimo (Dioscorea opposita Thunb.) tuber mucilage tororo: antioxidative activity and antihypertensive activity.

Yam tsukuneimo tuber mucilage tororo hydrolysates were prepared by autolysis and three different peptic enzymes. Except for pepsin hydrolysate, tororo was perfectly digested. Each hydrolysate for 100 mg/ml significantly prolonged the induction period of auto-oxidation of linoleic acid, which was similar to 5 mM ascorbic acid. These hydrolysates also possessed high scavenging activities such as superoxide anion radicals, hydroxyl radicals, and DPPH radicals. Moreover, high antihypertensive activities were detected in these hydrolysates except for autolysate, which were similar to various fermented foods such as miso, natto, sake, cheese, and so on. Present findings suggest that yam tsukuneimo tuber mucilage tororo may be useful for preventing diseases associated with reactive oxygen species and blood pressure in the body system and it can fully absorb the useful components from it to digest using the gastrointestinal enzymes.

Effects of tributyltin on the energy metabolism of pen shell (Atrina pectinata japonica).

We examined the effects of tributyltin (TBT) on aerobic and anaerobic energy metabolism of pen shell (Atrina pectinata japonica). We exposed pen shells to TBT at nominal concentrations of 0 (control) and 1.0microg/l for 72h under aerobic condition. At the end of the exposure, half of the pen shells in each treatment were wrapped in plastic wrap to simulate exposure to hypoxia and held at 25 degrees C for another 12h. The concentrations of the products of energy metabolism, namely lactate, pyruvate, fumarate and succinate, in adductor muscle were measured. The exposure to TBT under aerobic condition significantly elevated lactate, pyruvate and fumarate concentrations (p<0.001). After subsequent exposure to anaerobic condition, the mean concentration of succinate in the TBT treatment group was 64% of that in the control group, but there were no significant differences. Our results suggest that the energy metabolism of pen shell is disrupted by exposure to TBT.

Effect of tributyltin on veliger larvae of the Manila clam, Ruditapes philippinarum.

We investigated the effects of waterborne and maternal exposure to tributyltin (TBT) on veliger larvae of the Manila clam, Ruditapes philippinarum. In a waterborne exposure test, veliger larvae (D-larvae stage: 24h after fertilization) were exposed to TBT at measured concentrations of <0.01 (control), 0.055, 0.130, 0.340, and 0.600microg/l for 13d. The percentage of normal veliger larvae (the ratio of normal veliger larvae to all larvae) decreased significantly in all TBT treatment groups compared with that in the control group. In a maternal exposure test, 100 clams were exposed to TBT at measured concentrations of <0.01 (control), 0.061, and 0.310microg/l at 20-22 degrees C for 3 weeks, and the percentage of normal veliger larvae assessed for 13d. No maternal effects on veliger larvae from TBT were observed in TBT treatment groups as compared with the control group. These results demonstrate that waterborne TBT affects Manila clam veliger larvae, and indicates that TBT may have reduced Manila clam populations by preventing the development and survival of veliger larvae.

Tributyltin contamination of bivalves in coastal areas around northern Kyushu, Japan.

We determined tributyltin (TBT) concentrations in bivalve samples of blue mussel (Myitlus edulis), Manila clam (Ruditapes philippinarum) and pen shell (Atrina pectinata) collected from coastal areas around northern Kyushu in 1998 and 2001. TBT was detected in all bivalve samples collected, ranging in concentration from 0.008 to 0.135 microg/g wet wt. In Hakata Port, which is an industrial area, high TBT concentrations were detected in bivalves (blue mussel, maximum concentration of 0.135 microg/g wet wt). In the Ariake Sea, which is an important bivalve habitat, TBT concentrations in Manila clams ranged from 0.062 to 0.125 microg/g wet wt in 1998 and from 0.008 to 0.033 microg/g wet wt in 2001. In addition, concentrations of TBT in pen shells collected from the Ariake Sea in 2001 ranged from 0.009 to 0.095 microg/g wet wt. These results clearly demonstrate that, despite the regulation of TBT usage since 1990 in Japan, contamination of bivalves by TBT has persisted in coastal areas around northern Kyushu.

Effects of tributyltin maternal and/or waterborne exposure on the embryonic development of the Manila clam, Ruditapes philippinarum.

We examined the effect of tributyltin (TBT) on embryonic development of the Manila clam, Ruditapes philippinarum. In a maternal exposure test, 100 clams were exposed to TBT at measured concentrations of <0.01 (control), 0.061, 0.310, or 0.350 microg/l at 20-22 degrees C for 3 weeks, and the embryo developmental success (the ratio of normal D-larvae to all larvae) was measured. There was a significant negative correlation between embryo developmental success and TBT concentration in the female Manila clams (p < 0.001). These results indicated that TBT accumulated in the female clam decreased embryo developmental success. In a waterborne exposure test, fertilized eggs (4 h after fertilization) were exposed to TBT at measured concentrations of <0.01 (control), 0.062, 0.140, 0.320, or 0.640 microg/l for 23 h. Embryo developmental success was also significantly decreased in all TBT treatment groups compared with that in the control group. TBT accumulated in female adults and waterborne TBT clearly inhibit reproductive success of the clam.

Effect of maternal exposure to tributyltin on reproduction of the pearl oyster (Pinctada fucata martensii).

We examined the effect of tributyltin (TBT) on reproduction of the pearl oyster (Pinctada fucata martensii). In a maternal exposure test, five female pearl oysters were exposed to TBT at measured concentrations of 0 (control), 0.092, or 0.191 microg/L at 25 degreesC for one week, and the embryo developmental success (the ratio of normal D-larvae to all larvae) was measured. The embryo developmental success was significantly decreased in the 0.191-microg/L treatment group (65.5%) compared to that in the control group (82.5%; p = 0.031). Concentrations of TBT in the ovary reached 0.088 microg/g in the 0.191-microg/L treatment group. In a waterborne exposure test, inseminated eggs were exposed to TBT at measured concentrations of 0 (control), 0.020, 0.045, 0.091, 0.192, or 0.374 microg/L for 24 h. The embryo developmental success also was significantly decreased in the 0.192-microg/L treatment group (78.3%; p = 0.020) and no development at all was observed in the 0.374-microg/L treatment group compared with that in the control group (95.4%). These results clearly demonstrate that TBT accumulating in the bodies of bivalves has the potential to inhibit reproduction.