Purification and Characterization of a Novel Alginate Lyase from a Marine Streptomyces Species Isolated from Seaweed

Alginate, a natural polysaccharide derived from brown seaweed, is finding multiple applications in biomedicine via its transformation through chemical, physical, and, increasingly, enzymatic processes. In this study a novel alginate lyase, AlyDS44, was purified and characterized from a marine actinobacterium, Streptomyces luridiscabiei, which was isolated from decomposing seaweed. The purified enzyme had a specific activity of 108.6 U/mg, with a molecular weight of 28.6 kDa, and was composed of 260 amino acid residues. AlyDS44 is a bifunctional alginate lyase, active on both polyguluronate and polymannuronate, though it preferentially degrades polyguluronate. The optimal pH of this enzyme is 8.5 and the optimal temperature is 45 °C. It is a salt-tolerant alginate lyase with an optimal activity at 0.6 M NaCl. Metal ions Mn²⁺, Co²⁺, and Fe²⁺ increased the alginate degrading activity, but it was inhibited in the presence of Zn²⁺ and Cu²⁺. The highly conserved regions of its amino acid sequences indicated that AlyDS44 belongs to the polysaccharide lyase family 7. The main breakdown products of the enzyme on alginate were disaccharides, trisaccharides, and tetrasaccharides, which demonstrated that this enzyme acted as an endo-type alginate lyase. AlyDS44 is a novel enzyme, with the potential for efficient production of alginate oligosaccharides with low degrees of polymerization.     

Advanced Strategies for 3D Bioprinting of Tissue and Organ Analogs Using Alginate Hydrogel Bioinks

Alginate is a natural polysaccharide that typically originates from various species of algae. Due to its low cost, good biocompatibility, and rapid ionic gelation, the alginate hydrogel has become a good option of bioink source for 3D bioprinting. However, the lack of cell adhesive moieties, erratic biodegradability, and poor printability are the critical limitations of alginate hydrogel bioink. This review discusses the pivotal properties of alginate hydrogel as a bioink for 3D bioprinting technologies. Afterward, a variety of advanced material formulations and biofabrication strategies that have recently been developed to overcome the drawbacks of alginate hydrogel bioink will be focused on. In addition, the applications of these advanced solutions for 3D bioprinting of tissue/organ mimicries such as regenerative implants and in vitro tissue models using alginate-based bioink will be systematically summarized.     

A comparison of the use of different swab materials for optimal diagnosis of amoebic gill disease (AGD) in Atlantic salmon (Salmo salar L.)

Routine gill swabbing is a non-destructive sampling method used for the downstream qPCR detection and quantitation of the pathogen Neoparamoeba perurans, a causative agent of amoebic gill disease (AGD). Three commercially available swabs were compared aiming their application for timelier AGD diagnosis (Calgiswab^® (calcium alginate fibre-tipped), Isohelix^® DNA buccal and cotton wool-tipped). Calcium alginate is soluble in most sodium salts, which potentially allows the total recovery of biological material, hence a better extraction of target organisms’ DNA. Thus, this study consisted of (a) an in vitro assessment involving spiking of the swabs with known amounts of amoebae and additional assessment of retrieval efficiency of amoebae from agar plates; (b) in vivo testing by swabbing of gill arches (second, third and fourth) of AGD-infected fish. Both in vitro and in vivo experiments identified an enhanced amoeba retrieval with Calgiswab® and Isohelix® swabs in comparison with cotton swabs. Additionally, the third and fourth gill arches presented significantly higher amoebic loads compared to the second gill arch. Results suggest that limiting routine gill swabbing to one or two arches, instead of all, could likely lead to reduced stress-related effects incurred by handling and sampling and a timelier diagnosis of AGD.     

Dietary supplementation with low molecular weight sodium alginate improves growth,haematology, immune reactions and resistance against Aeromonas hydrophila in Clarias gariepinus

A few studies have illustrated the effects of sodium salt derived from alginic acid on different fish species. However, little is known about the effect of sodium alginate on catfish (Clarias gariepinus). Therefore, this study was performed to assess the use of low molecular weight sodium alginate ( LMWSA) in C. gariepinus. A total of 180 apparently healthy C. gariepinus with a mean body weight of 45 g were randomly divided into three equal groups (D1, D2 and D3). D1 the control group received a control diet, while D2 and D3 received 1% and 3% LMWSA, respectively, for 8 weeks. A challenge test against Aeromonas hydrophila was performed on 15 randomly selected catfish for 15 days. At the end of the experiment, catfish in D3 that received a diet of 3% LMWSA showed significant increases in the final body weight, weight gain and thermal‐unit growth coefficient compared with those in D2 and D1. There was a significant decrease in the erythrogram in D1 after the 4‐day pathogen challenge. A leucogram revealed leucocytosis, heterophilia and lymphocytosis in catfish in D2 and D3 compared with those in D1. After the 4‐day challenge, the following changes took place: lysozyme, nitric oxide, phagocytic activity and the respiratory burst were significantly elevated in catfish that received LMWSA and were more pronounced in D3 than in D1. The mortalities of catfish have been stopped after pathogen challenge from 8‐day in D1 and D2 where at 6‐day in D3. Thus, administration of 1% and 3% LMWSA enhances the growth, immune response and resistance of C. gariepinus against A. hydrophila.     

Growth stimulation of juvenile abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> by feeding with salmon growth hormone in sodium alginate gel

We examined the ability of salmon growth hormone (sGH) in sodium alginate (SA) gel to accelerate the somatic growth of juvenile abalone, Haliotis discus hannai, by feeding. After the feeding of sGH at 50 μg or 100 μg in 350 mg SA gel, immunoreactivity to sGH in body fluid was maximal after 12 h, and was still detectable at 24 h. No immunoreactivity was observed in the control group. Following the feeding of sGH at 0.5 mg or 5 mg/8 g of SA gel at 7- and 14-day intervals, there was a greater increase in shell length and body weight than in the control. In abalone, sGH can be transported from food into the circulatory system and subsequently improve somatic growth.     

兰州百合粗低聚糖对嗜热链球菌ATCC-14485和保加利亚乳杆菌ATCC-11842产酸及增殖的影响

益生元在促进益生菌生长、调节肠道微生态平衡方面发挥着重要作用。为探究兰州百合粗低聚糖（crude lily oligosaccharides, CLOs）的益生元特性,以兰州百合干为原料,提取、表征CLOs;以低聚果糖和菊粉作为阳性对照,测定CLOs在人工胃肠溶液的水解度,以及对嗜热链球菌ATCC-14485和保加利亚乳杆菌ATCC-11842的产酸、增殖作用的影响。结果表明：CLOs分子量为1 194 Da,主要由甘露糖、阿拉伯糖和葡萄糖组成,摩尔比为1:3.57:16.40;CLOs经人工胃肠溶液消化后水解度是26.96%,显著低于菊粉（P <0.05）;CLOs促进乳酸菌发酵产酸的效果显著优于菊粉和低聚果糖（P <0.05）;当CLOs的添加量为1.0%(W/V)时,能有效促进嗜热链球菌和保加利亚乳杆菌的增殖,并减缓二者在货架期的死亡速率。研究结果说明CLOs具有益生元特性,可为新型共生发酵乳的研发提供理论依据。     

二氢吡啶海藻酸钙凝胶小球的制备工艺

研究二氢吡啶海藻酸钙凝胶小球的制备工艺和最优处方.以海藻酸钠为基质材料,采用滴制法制备海藻酸钙凝胶小球,以二氢吡啶的包封率和载药量作为制备工艺优化指标,设计正交试验优选最佳处方;测定最佳处方所制制剂的包封率(Encapsulation efficiency,EE)和载药量(Drug loading,LD)并与原料药比较体外释放作用.最优处方为海藻酸钠浓度1.5%,海藻酸钠与二氢吡啶质量比1:2,氯化钙浓度0.10 mol/L;EE和LD值分别为81.16%,48.20%,与原料药相比缓释性较好,且具有良好的溶胀性能.本法工艺简单、可行、稳定,重现性好.     

基于酶技术生产低黏度及超低黏度褐藻胶的工艺

为解决当前低黏度及超低黏度褐藻胶生产工艺中存在的不足,本实验以海带为原料,利用褐藻胶裂解酶降解制备低黏度及超低黏度褐藻胶,研究了分子量、p H、温度对黏度的影响,确定了碱消化的最佳条件,探究了酶解工艺中加酶量、酶解时间及原料的初始黏度对褐藻胶产品的影响。结果显示,通过控制加酶量(100~500 U/g),酶解30 min即可得到低黏度及超低黏度褐藻胶,其中加酶量为100~330 U/g时可得到低黏度褐藻胶,加酶量增加至330~500 U/g时,可得到超低黏度褐藻胶,且酶解法得到的褐藻胶样品分子量均一度高,工艺节水率高达10%~50%;同时研究发现酶解样品黏度与原料初始黏度相关性不大,只在较短时间内表现出相关性,该工艺具有较高的原料适用性。     

金龟子绿僵菌的藻胶颗粒剂型及其促进种群恢复作用

为了保护绿僵菌的存活能力并促进其田间增殖,本试验研究了以海藻胶粒剂型化金龟子绿僵菌的成胶浓度、载体、促活组分、外膜等因素对绿僵菌的存活保护与增殖促进作用。结果确定了适合于金龟子绿僵菌与海藻酸钠、钙离子交联反应的最适条件为海藻酸钠2%、氯化钙0.2~0.3 mol/L;填充4%~10%凹凸棒土支撑颗粒内部胶网,有利于孢子存活和恢复生长;添加0.5%玉米粉或糊精作为促活剂,可使孢子增殖率提高60%;以1.5%海藻酸钠与3.5%聚乙烯醇联合形成的颗粒剂外膜对抵御外界微生物侵袭有一定作用。     

含茉莉酸甲酯的生物活性膜对蓝莓采后保鲜效果的研究

以蓝莓为试材,采用涂膜与生物防治结合的方法,以1.5%海藻酸钠+0.3%甘油为主要材料,无菌水为溶剂,分别加入109CFU/m L罗伦隐球酵母(Cryptococcus laurentii)和100μmol/L茉莉酸甲酯进行涂膜处理,研究蓝莓在0~2℃下贮藏28 d的保鲜效果。结果表明,采用1.5%海藻酸钠+0.3%甘油+109CFU/m L C.laurentii+100μmol/L茉莉酸甲酯复合涂膜液处理的蓝莓失重率和腐烂指数明显降低,糖、酸和花青素含量的变化减缓,果实可保持较高的硬度和弹性,丙二醛的产生受到抑制,而超氧化物歧化酶活性得到提高,使蓝莓果实能够保持良好的外观品质和较高含量的营养成分,保鲜效果最佳。