Purification of serine carboxypeptidase from the hepatopancreas of Japanese common squid Todarodes pacificus and its application for elimination of bitterness from bitter peptides

ABSTRACT:   An acidic serine carboxypeptidase (CPase Tpa) from the hepatopancreas of Japanese common squid Todarodes pacificus was purified. Purified CPase Tpa had a molecular mass of 36 kDa on sodium dodecylsulfate–polyacrylamide gel electrophoresis, and an isoelectric point of 6.0. The optimum pH of CPase Tpa was pH 4.0. In investigating the specificity of CPase Tpa for several peptide substances, it was found that peptides with hydrophobic or bulky amino acid residues at the P₁ position reacted well. The enzymatic activity was almost completely inhibited by p -chloromercuribenzoic acid, monoiodoacetic acid, diisopropylfluorophosphate and HgCl₂. This is the basis for its grouping in the serine carboxypeptidase family (EC 3. 4. 16. 5). The substrate specificity of CPase Tpa can be used to eliminate the bitterness of bitter peptides. In this study, the bitterness-reductive effect using bitter peptides prepared by hydrolyzing soy protein, casein and corn gluten with pepsin or trypsin was tested. The bitterness of soy peptide digested with pepsin was completely eliminated by treatment with CPase Tpa, whereas the bitterness of casein digested with trypsin and corn peptide digested with pepsin were somewhat less efficient. On the basis of these results, it is anticipated that CPase Tpa would be effective in eliminating the bitterness of some bitter peptides.     

Density and sound speed contrasts of the Japanese common squid Todarodes pacificus and their influence on acoustic target strength

ABSTRACT:   In this study, density and sound speed contrasts were measured for the Japanese common squid Todarodes pacificus . Target strength (TS) data derived from an acoustic scattering model based on measurements of these two parameters were compared with TS values based on acoustic measurement data to determine whether the measured parameters are reasonable values for theoretical scattering model. Density contrast ( g ) was measured from the displacement volume and wet weight, while sound speed contrast ( h ) was measured from the acoustic measurements of travel time (time-of-flight method). The Kirchhoff ray mode model, which represents the squid body as a set of fluid-filled cylinders, was used to calculate theoretical TS. Mean g- and h -values of the squid were 1.029 and 1.041, respectively. Comparison with previous data showed that g was similar, whereas h was relatively high. The difference in the TS between the theoretical model using measured parameters and the acoustic measurement was within 3 dB. Thus, the measured g- and h -values are acceptable for theoretical models of squid.     

Effect of water temperature on gonadal development and emaciation of Japanese common squid <Emphasis Type="Italic">Todarodes pacificus</Emphasis> (Ommastrephidae)

ABSTRACT:   In this study, the site of spawning ground and the relationship between oceanographic conditions and gonadal development and emaciation of the Japanese common squid Todarodes pacificus were examined based on data from research cruises. The research cruises were conducted in the south-western part of the Sea of Japan and the East China Sea in the winter of 1999. Sea surface temperature (SST) ranged 12.7–23.2°C in the survey area. Mature females concentrated on the continental shelf of the East China Sea, suggesting a major spawning ground was formed in this area. Gonadal development of the collected squid was examined using ovary somatic index (OSI) and oviduct somatic index (ODSI) in females, and emaciation was examined using mantle muscle somatic index (MI) and hepatosomatic index (HSI). Mean OSI and ODSI increased with SST at the stations where SST was greater than 15°C, particularly where SST ranged between 15 and 18°C. In contrast, mean MI and HSI decreased with increase in SST at the stations where SST was above 15°C. These results indicate that female gonads develop rapidly at SST above 15°C, and at the same time adult squids tend to be emaciated at this temperature condition.     

海洋环境因子对日本海太平洋褶柔鱼渔场时空分布的影响

根据2010—2013年11—12月日本海太平洋褶柔鱼(Todarodes pacificus)的渔获生产数据,并结合遥感获取的海洋环境数据,利用渔获量重心法、地统计插值和数理统计方法,分析了太平洋褶柔鱼的资源丰度与渔获量重心的时空变化及其与主要环境因子(海表温度、叶绿素浓度、海流)之间的关系。研究表明,渔场重心每年稍有差异,经度重心集中在132°20′E附近;纬度重心集中在36°30′N和37°30′N两个位置附近。GAM模型显示,日本海太平洋褶柔鱼渔场的最适海表温度为16~18℃;最适叶绿素a浓度为0.37~0.45 mg/m3。海洋环境与单位捕捞努力渔获量(CPUE)的回归方程的显著性检验表明,除了叶绿素a浓度呈一般显著外,海表温度和空间因子在CPUE上的回归均极显著(P0.01),符合统计意义。4年间渔场的适宜环境范围有所差异,推断主要是日本海海域对马暖流、东朝鲜暖流与里曼寒流相互交汇的强弱作用力引起的,也有不同年份季风的强弱不同以及全球气候变化的因素存在。每一年的渔场海洋环境因子稍有差别,对其资源量的影响巨大。     

Target strength measurements of suspended live ommastrephid squid, <Emphasis Type="Italic">Todarodes pacificus</Emphasis>, and its application in density estimations

ABSTRACT:   Average target strength (TS) measurements of free-swimming Todarodes pacificus are required for calculating abundance estimations using the echo integration method. In this study, 31 live T. pacificus (18.0–28.4 cm dorsal mantle length; [ML]) were suspended with mantle tilt angles horizontally similar to free-swimming orientation, and their TS were measured using a 38 kHz split-beam echo sounder. Normalized TS by squared ML of T. pacificus ML was calculated as −73.1 dB: TS = 20 log ML −73.1 dB. The density of T. pacificus at their fishing ground off the northern Sanriku coast, Japan, was then estimated first using the echo integration method with the obtained normalized TS measurements, and then using commercial purse seine fishery CPUE (catch (tons) of squid per haul of purse seine) data. The density of T. pacificus aggregations near the continental shelf edge and upper continental slope was estimated as approximately 50–400 individuals/100 m² using both the echo integration method and CPUE data, both density results were similar. It was considered feasible therefore, to assess T. pacificus biomass directly using the echo integration method.     

Distribution and migration of Japanese common squid, Todarodes pacificus, in the southwestern part of the East (Japan) Sea

Data on squid catches of Korean and Japan, water temperatures at depth of 100 m, and night-visible images of fishing boats collected in the East (Japan) Sea from 1970 to 1999 were analyzed to examine the distribution and migration of the Japanese common squid, Todarodes pacificus, in the southwestern part of the East (Japan) Sea. The main fishing grounds detected from squid catch in each grid (0.5° latitude × 0.5° longitude) by Korean squid fishery and night-visible images provided by the Defense Meteorological Satellite Program (DMSP) Operational Linescan System (OLS) were situated in the southwestern part of the East (Japan) Sea. The distribution and migration route was illustrated from squid catch in each grid and DMSP OLS image. In years of high catches, the fishing grounds were situated mainly between Ulleung Island and the eastern coastal waters of Korea, while in years of low catches they were situated between Ulleung Island and the Yamato Bank in the central East (Japan) Sea. The center of fishing activity began moving northward from around the Korea/Tsushima strait to the northern boundary of the Tsushima Warm Current in March, reaching into the Yamato Bank in September, and then returning to the strait by February. The northward and southward migration routes differed; the northward migration route occurred closer to the mainland coast of Korea than the southward migration route did. This work suggests T. pacificus begin their northward migration almost 2 months earlier than previously suggested.     

Anionic Trypsin from the Pyloric Ceca of Pacific Saury (Cololabis saira): Purification and Biochemical Characteristics

Anionic trypsin from Pacific saury (Cololabis saira) pyloric ceca was purified to homogeneity by ammonium sulfate precipitation, ion-exchange chromatography, and gel filtration chromatography. It was purified to 53.7-fold with a yield of 6.1%. The apparent molecular weight of the enzyme was about 24 kDa, as determined by size exclusion chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). On native-PAGE, trypsin showed a single band. The purified anionic trypsin displayed optimal activity at pH 8.5 and 55°C. The enzyme was stable at neutral and alkaline pH and in the temperature range of 20–50°C. The stability was affected by the calcium ion. The activity of purified anionic trypsin was completely inhibited by soybean trypsin inhibitor and N-p-tosyl-L-lysine chloromethyl ketone (TLCK) and partially inhibited by ethylenediaminetetraacetic acid (EDTA). NaCl (0–30%) decreased the activity in a concentration-dependent manner. The kinetic trypsin constants K_m and K_cat were 0.19 mM and 210 s^?1, respectively, while the catalytic efficiency (K_cat/K_m) was 1105.26 s^?1 mM^?1. The N-terminal amino acid sequences of anionic trypsin, IVGGYECQAH, were found and were homologous to those of trypsin from other fish species.     

Cloning of estrogen receptor-β cDNA of common Japanese conger Conger myriaster and expression of the gene during oocyte development

ABSTRACT:   Estrogen receptor (ER) cDNA was cloned from the liver of common Japanese conger Conger myriaster . Common Japanese conger ER cDNA contains 2543 nucleotides including an open reading frame that encodes 596 amino acids, with a molecular weight of 66.1 kDa. The deduced amino-acid sequence of the common Japanese conger ER shares 85% identity with that of eel Anguilla japonica ER-β, and 39–46% and 53–59% identity with the deduced sequences of ER-α and ER-β of other species, respectively. In a phylogenetic analysis, the common Japanese conger ER was found to cluster with the ER-β of other species. The common Japanese conger ER-β also has six domains (A–F), which are conserved in the nuclear receptor superfamily. The amino-acid sequences in the C and E domains were highly conserved between common Japanese conger ER-β and other ER. However, the other domains showed low homology. In the female common Japanese conger, ER-β mRNA was highly expressed in the liver and pituitary gland. The expression levels of the ER-β gene increased from the oil droplet stage to the primary yolk globule stage and were maintained until the migratory nucleus stage.     

Lipid content and fatty acid composition of muscle,liver, ovary and eggs of captive-reared and wild silver Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> during artificial maturation

ABSTRACT:   Changes in lipid content and fatty acid composition of muscle, liver and ovary of captive-reared and wild silver Japanese eel Anguilla japonica were examined during artificial maturation induced by salmon pituitary homogenate (SPH) injections. Although the relative levels of n-3 and n-6 highly unsaturated fatty acids (HUFA) in liver and ovary were higher than in muscle in both captive and wild silver eels before SPH injection, these tended to decrease with maturation. The relative levels of n-6 HUFA in muscle, liver, ovary and eggs of wild silver eels were remarkably higher than those in captive eels. Therefore, we attempted to alter the ratio of n-6 HUFA in eggs by feeding eels a diet supplemented with linoleic acid-rich plant oil. Although the percentage of n-6 polyunsaturated fatty acids to total fatty acids in eggs of eels fed the supplemented diet was similar to wild silver eels, the percentage of n-6 HUFA remained remarkably lower than in wild silver eels. Hence, it appears that the supplemented diet affected the fatty acid composition of eggs, but did not result in much conversion of linoleic acid to its higher homologs in eels.     

Mitochondrial DNA sequence variation of Japanese anchovy <Emphasis Type="Italic">Engraulis japonicus</Emphasis> from the Yellow Sea and East China Sea

Sequence variation of a fragment of the mitochondrial cytochrome b (cyt b) and cytochrome c oxidase subunit I (COI) gene was examined using polymerase chain reaction and direct sequencing among three populations of Japanese anchovy Engraulis japonicus from the Yellow Sea and East China Sea. A total of 24 and 47 nucleotide sites were detected variable defining 29 and 32 haplotypes in cyt b and COI data, respectively. All variable sites except one in COI were silent in the two sets of sequences. The Ewens-Watterson test indicated that the observed allelic configurations in both data sets were in full agreement with neutral expectations. The variation level was high, with h=0.957±0.018, π=0.644±0.387 (%) in cyt b data set and h=0.958±0.021, π=0.640±0.386 (%) in COI data set, respectively. However, at the population level, Fst values between pairs of populations were not significantly different from zero (P>0.05) in both data sets. The analysis of haplotype frequency distribution showed no significant differences among populations. Similarly, the analysis of the partitioning of molecular variance indicated that all or almost all of the genetic variation was distributed within populations. Based on the data from this study, the existence of separate genetic stocks in this area were not detected. Mixing of stocks to some extent in migration cycle and dispersal capacity of anchovy’s planktonic larvae could be the reasons for genetic homogeneity in this species in the Yellow Sea and East China Sea.