A Novel Gelatinase from Marine Flocculibacter collagenilyticus SM1988: Characterization and Potential Application in Collagen Oligopeptide-Rich Hydrolysate Preparation

Although the S8 family in the MEROPS database contains many peptidases, only a few S8 peptidases have been applied in the preparation of bioactive oligopeptides. Bovine bone collagen is a good source for preparing collagen oligopeptides, but has been so far rarely applied in collagen peptide preparation. Here, we characterized a novel S8 gelatinase, Aa2_1884, from marine bacterium Flocculibacter collagenilyticus SM1988^T, and evaluated its potential application in the preparation of collagen oligopeptides from bovine bone collagen. Aa2_1884 is a multimodular S8 peptidase with a distinct domain architecture from other reported peptidases. The recombinant Aa2_1884 over-expressed in Escherichia coli showed high activity toward gelatin and denatured collagens, but no activity toward natural collagens, indicating that Aa2_1884 is a gelatinase. To evaluate the potential of Aa2_1884 in the preparation of collagen oligopeptides from bovine bone collagen, three enzymatic hydrolysis parameters, hydrolysis temperature, hydrolysis time and enzyme-substrate ratio (E/S), were optimized by single factor experiments, and the optimal hydrolysis conditions were determined to be reaction at 60 ℃ for 3 h with an E/S of 400 U/g. Under these conditions, the hydrolysis efficiency of bovine bone collagen by Aa2_1884 reached 95.3%. The resultant hydrolysate contained 97.8% peptides, in which peptides with a molecular weight lower than 1000 Da and 500 Da accounted for 55.1% and 39.5%, respectively, indicating that the hydrolysate was rich in oligopeptides. These results indicate that Aa2_1884 likely has a promising potential application in the preparation of collagen oligopeptide-rich hydrolysate from bovine bone collagen, which may provide a feasible way for the high-value utilization of bovine bone collagen.     

鳄鱼血蛋白酶解产物抗氧化特性和血管紧张素转化酶抑制活性研究

研究了鳄鱼血蛋白酶解产物的抗氧化特性和对血管紧张素转化酶（ ACE）的抑制活性。利用木瓜蛋白酶酶解鳄鱼血浆蛋白和血球蛋白,用分光光度法测定了酶解产物的抗氧化能力和用高效液相色谱（ HPLC）测定其ACE的抑制率。结果显示：鳄鱼血浆和血球蛋白酶解产物的亚铁离子螯合能力差异性不显著（ P＞0．05）;在0～5 mg/mL的浓度范围内,血球蛋白酶解产物清除ABTS自由基的能力大于血浆蛋白酶解产物,且在浓度为1 mg/mL时,两者清除ABTS自由基的能力差异性极显著（ P＜0．01）;血浆蛋白酶解产物清除DPPH自由基的能力在0～5 mg/mL的浓度范围内随着蛋白浓度的增加而升高,血球蛋白酶解产物在蛋白浓度为4 mg/mL处达到最大清除率,之后下降;在0～20 mg/mL的浓度范围内,两种酶解产物的还原力随着蛋白浓度的提高显著升高,但两者还原力的差异性不显著（ P＞0．05）;鳄鱼血浆和血球蛋白酶解产物对ACE具有良好的抑制力,其最大抑制率可分别达到75．56％和86．42％。研究表明,鳄鱼血蛋白酶解产物在体外具有抗氧化和抑制ACE的活性。     

Taurine alone or in combination with fish protein hydrolysate affects growth performance,taurine transport and metabolism in juvenile turbot (Scophthalmus maximus L.)

The study was conducted to investigate the effects of taurine (Tau) alone or in combination with fish protein hydrolysate (FPH) on growth performance, the expression of Tau transporter (TauT) and metabolic profile in juvenile turbot. FM, FPH0, FPH0+T, FPH10 and FPH10+T diets, respectively, contained 300, 150, 150, 80, and 80 g/kg fishmeal. FPH10 and FPH10+T diets contained 62 g/kg FPH. FPH0+T and FPH10+T diets were, respectively, prepared by supplementing the FPH0 and FPH10 diet formulations with 8 g/kg Tau. Specific growth rate was the highest in FM group and the lowest in FPH10 group. TauT mRNA levels in fish fed Tau supplemented diets were significantly lower than that in Tau unsupplemented diets. NMR‐based metabolomics analysis showed that Tau contents in liver of FPH0+T and FPH10+T were significantly higher than that of FM, FPH0 and FPH10. In muscle, Tau contents were significantly decreased in the FPH10+T versus FPH0 and the FPH10+T versus FPH10 comparisons. In conclusion, 62 g/kg FPH to replace fishmeal may not affect Tau synthesis, transport and metabolism. However, Tau supplemented alone or in combination with a certain level of FPH could reduce the requirement for Tau synthesis and transport and increased Tau levels in muscle and liver.     

Production of Interesting Peptide Fractions by Enzymatic Hydrolysis of Tuna Dark Muscle By-Product Using Alcalase

ABSTRACT

A protein hydrolysate was prepared from proteins of tuna dark muscle by-product. The hydrolysis conditions (time, temperature, pH, and enzyme concentration) using Alcalase was optimized by response surface methodology (RSM). The regression coefficient close to 1.0, observed during experimental and validation runs, indicated the validity of the model. The hydrolysate produced under the optimum conditions determined by RSM has a low rate of peptide fraction of molecular weight of 4–1 kDa. Meanwhile, the results obtained by hydrolysis under optimal conditions determined by a complementary study (temperature 55°C, time 60 min, 1% enzyme concentration, and pH 8.5) show that the hydrolysate produced has a height rate of the peptide fraction of molecular weight of 4–1 kDa. The amino acid composition of the protein hydrolysate prepared proved to have the potential for application as an ingredient in balanced fish diets and as a source of nitrogen in microbial growth media.     

Tuna viscera protein hydrolysate: nutritive and disease resistance properties for Persian sturgeon (Acipenser persicus L.) larvae

The effect of fish protein hydrolysate (FPH) on the growth, feed utilization, digestive enzymes, gut microflora and resistance to bacterial challenge in Persian sturgeon (Acipenser persicus) larvae during a 45‐day feeding trial has been investigated. Artificial diets containing different levels of tuna visceral protein hydrolysate (PH0, PH10, PH25 and PH50) and live foods (Artemia: 5 days, Daphnia: 40 days) were fed to Persian sturgeon larvae. Larvae fed on the PH10 and PH25 had growth, feed utilization, protein content and trypsin activity significantly better than PH0 and PH50 groups (P < 0.05). Condition factors, survival rate and pepsin activity were not significantly different (P > 0.05). Brush border membrane enzymes to cytosolic enzyme ratio revealed that maturation had been occurred in 41 days post hatch (dph) at moderate FPH levels. In 54 dph larvae, this ratio was not affected by the level of FPH in the diet. No significant differences were observed in the total number of gut microflora or in the resistance to Aeromonas hydrophila (P > 0.05), whereas lactic acid bacteria was significantly higher in PH50 (P < 0.05). The results of this study show that application of moderate levels of FPH in Persian sturgeon diets could enhance larvae performance due to the early maturation in intestinal.     

Sensory Characteristics of Threadfin Bream (Nemipterus japonicus) Hydrolysate

ABSTRACT

Threadfin bream (Nemipterus japonicus) spray-dried hydro-lysate was prepared using Alcalase at predetermined optimum hydrolysis conditions consisting of pH 8.5, 60°C, 2% enzyme, and 2 h hydrolysis. The hydrolysate was evaluated for its preferred concentration in rice porridge. Its acceptability was compared with sauces and enzyme-treated rice and vegetable hydrolysate. The preferred concentration in rice porridge was found to be 3% (w/v) and bitterness increased with concentration of hydrolysate. Threadfin bream hydrolysate was preferred as much as soy sauce; however, it received a lower score for taste compared to fish sauce. In terms of color, threadfin bream hydrolysate received significantly (p < 0.05) lower score than Flavorzyme and Alcalase treated rice hydrolysate and Vegetable hydrolysate. Low preference to the hydrolysate could probably be due to its greenish yellow color in which the common color for flavorants are brownish. For overall acceptability, no significant difference was observed between threadfin bream hydrolysate and Alcalase-treated rice hydrolysate but was significantly lower than vegetable hydrolysate and Flavorzyme-treated rice hydrolysate.

The hydrolysis of threadfin bream with Alcalase produced hydrolysate having a combination of bitter, umami, salty, sweet and sour tastes with fishy flavor. Bitterness and umami were rated as having an intermediate intensity, salty as between weak and intermediate, and sour and sweet as weak. The three basic tastes for hydrolysate were bitter, umami and salty, with umami and bitterness being the determining tastes.     

Purification and Characterization of a New DPPH Radical Scavenging Peptide from Shrimp Processing By-products Hydrolysate

A peptide with high antioxidant activity was isolated and identified from shrimp processing by-products hydrolysate. The 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity was used to evaluate the antioxidant activity of fractions. The purified antioxidant peptide was identified as Ser-Val-Ala-Met-Leu-Phe-His (804.4 Da) by electrospray ionization tandem mass spectrometry. The purified peptide at 50 μg/mL showed antioxidative activity of 65.7 ± 0.9%, which was 3.18-fold higher compared with the first step separation by ion-exchange chromatography. It is possible to produce natural antioxidative peptides from shrimp processing by-products hydrolysate. The high antioxidant activity may be due to the presence of Phe-His segment at the C-terminus of the peptide.     

海藻多糖用于食品涂膜保鲜的研究现状

海藻多糖具有抑菌、抗病毒和抗氧化等多种生物活性,可用于食品的涂膜保鲜。本文概述了海藻多糖的生物活性及其保鲜机理,并将目前已发现的具有活性的海藻多糖种类及其在食品涂膜保鲜中的应用现状、保鲜效果等进行综述,同时对其未来发展趋势进行展望。     

日粮中鱼蛋白水解物对黄颡鱼生长、体成分和血清生理指标的影响

为了探讨鱼蛋白水解物对黄颡鱼生产性能的影响,以日本鳀粉为对照,以实用型黄颡鱼饲料配方模式为基础开展实验:1以30.5%鱼粉为对照(FM),在相同配方模式下,以6%鱼蛋白水解物(MPH6)替代20%的鱼粉;2以30.5%鱼粉为对照(FM),在无鱼粉日粮中分别添加3%(FPH3)、6%(FPH6)、12%(FPH12)鱼蛋白水解物;共设计5组等氮等能实验日粮,在池塘网箱中养殖黄颡鱼[初始体质量(30.08±0.35)g]60 d。结果显示:与FM相比,FPH12在SGR、FCR、PRR和FRR方面均无显著差异,而MPH6、FPH3、FPH6组SGR降低了15.45%~24.39%,FCR升高了32.14%~42.86%,MPH6、FPH6差异显著,在PRR和FRR方面,MPH6、FPH3、FPH6组PRR降低了21.11%~27.78%,MPH6组FRR降低了41.51%;全鱼水分、粗蛋白、粗脂肪和灰分各组间差异不显著,FPH3、FPH6、FPH12肌肉多种游离氨基酸水平显著高于FM,其中Thr、Val、His与其在日粮中的水平显著相关;FPH6组HSI显著低于FM,鱼蛋白水解物对CP、VSI、肠体比的影响不显著;血清AST、ALT、HDL、LDL、TP、CHOL、TG无显著差异,FPH3组ALB显著低于FM。研究表明:黄颡鱼日粮中,12%鱼蛋白水解物(干物质)与30.5%鱼粉在生长速度、饲料效率、血清生理指标等方面具有一定的等效性;过高的植物蛋白日粮影响了黄颡鱼的生产性能;饲喂鱼蛋白水解物日粮使黄颡鱼肌肉游离氨基酸的含量升高,特别是呈味氨基酸的含量增加。     

固定化细胞发酵半纤维素水解液产木糖醇的研究

固定在海藻酸钙凝胶中的热带假丝酵母(Candida tropicalis)细胞，可有效地利用玉米芯半纤维素水解液生产木糖醇。在摇瓶条件下，采用分批发酵方式，确立了适宜的发酵工艺参数为：32℃，每升氮源含酵母粉2．2g、胰蛋白胨3．7g，发酵液初始pH值6．0，分段改变摇床转速，其中0～24h为210r／min；24～72h为140r／min，固定化细胞凝胶珠与玉米芯半纤维素水解液体积比为1：4。利用固定化细胞重复进行10批次共30d发酵，木糖醇得率平均为73．7％，达到了理论值的80％。固定化细胞密度高、抗逆性强、发酵性能稳定，水解液未经脱色与离子交换便可转化成木糖醇，显示了良好的应用前景。