固定化生物硅藻土强化CAST工艺处理含盐污水研究

采用9%聚乙烯醇+0.5%海藻酸钠包埋耐盐菌群的固定化生物硅藻土小球投加到循环式活性污泥(CAST)反应器中,对比传统CAST工艺和投加固定化小球的CAST工艺对含盐污水COD和氨氮的去除效果,考察固定化生物硅藻土小球强化CAST工艺处理含盐污水性能。试验结果表明:在相同运行条件下,投加固定化生物硅藻土小球的CAST工艺出水COD、氨氮去除率基本维持在88%和90%以上,与传统CAST工艺相比分别提高了15%和10%。同时,向CAST工艺添加固定化生物硅藻土有助于维持反应器中微生物浓度稳定,提高出水水质及其稳定性。     

Bioactive properties of enzymatic hydrolysates from abdominal skin gelatin of yellowfin tuna (Thunnus albacares)

Gelatin (90.6 ± 0.1%) was optimally prepared by response surface methodology from yellowfin tuna (Thunnus albacares, YT) abdominal skin. To investigate bioactive properties of enzymatic hydrolysates from the abdominal skin gelatin (ASG), ASG was hydrolysed with alcalase, protamex, neutrase and flavourzyme as affected by hydrolysis time. Antioxidant, nitrite scavenging and angiotensin‐I converting enzyme (ACE) inhibitory activities of the hydrolysates were determined. Antioxidant activities of the hydrolysates were found through linoleic acid peroxidation inhibitory effects. Alcalase‐derived hydrolysates (AHs) were more effective than others in metal ions chelating, superoxide anion scavenging and hydroxyl radical scavenging activities (P < 0.05). AHs showed significantly stronger nitrite scavenging activities (44.4–60.7%) than others (P < 0.05). Fraction A from AH showed strong ACE inhibitory activity (IC₅₀ of 0.75 mg mL^?1). These results suggest that YT ASG and its enzymatic hydrolysates could be functional food and/or pharmaceutical ingredients with potent antioxidant, anticarcinogenic and antihypertensive benefits.     

Covalent Linkage of an R-ω-Transaminase to a d-Amino Acid Oxidase through Protein Splicing to Enhance Enzymatic Catalysis of Transamination

R-ω-Transaminases (RTAs) catalyse the conversion of R-configured amines [e.g., (R)-1-phenylethylamine] into the corresponding ketones (e.g., acetophenone), by transferring an amino group from an amino donor [e.g., (R)-1-phenylethylamine] onto an amino acceptor (e.g., pyruvate), resulting in a co-product (e.g., d -alanine). d -Alanine can be deaminated back to pyruvate by d -amino acid oxidase (DAAOs). Here, through in vivo subunit splicing, the N terminus of an RTA subunit (RTA^S) was specifically ligated to the C terminus of a DAAO subunit (DAAO^S) through native peptide bonds (RTA&DAAO). RTA^S is in close proximity to DAAO^S, at a molecular-scale distance. Thus the transfer of pyruvate and d -alanine between RTA and DAAO can be directional and efficient. Pyruvate→d -alanine→pyruvate cycles are efficiently formed, thus promoting the forward transamination reaction. In a different, in vitro noncovalent approach, based on coiled-coil association, the RTA^S N terminus was specifically associated with the DAAO^S C terminus (RTA#DAAO). In addition, the two mixed individual enzymes (RTA+DAAO) were also studied. RTA&DAAO has a shorter distance between the paired subunits (RTA^S–DAAO^S) than RTA#DAAO, and the number of the paired subunits is higher than in the case of RTA#DAAO, whereas RTA+DAAO cannot form the paired subunits. RTA&DAAO exhibited a transamination catalysis efficiency higher than that of RTA#DAAO and much higher than that of RTA+DAAO.     

Crossing the Border: From Keto- to Imine Reduction in Short-Chain Dehydrogenases/Reductases

The family of NAD(P)H-dependent short-chain dehydrogenases/reductases (SDRs) comprises numerous biocatalysts capable of C=O or C=C reduction. The highly homologous noroxomaritidine reductase (NR) from Narcissus sp. aff. pseudonarcissus and Zt_SDR from Zephyranthes treatiae, however, are SDRs with an extended imine substrate scope. Comparison with a similar SDR from Asparagus officinalis (Ao_SDR) exhibiting keto-reducing activity, yet negligible imine-reducing capability, and mining the Short-Chain Dehydrogenase/Reductase Engineering Database indicated that NR and Zt_SDR possess a unique active-site composition among SDRs. Adapting the active site of Ao_SDR accordingly improved its imine-reducing capability. By applying the same strategy, an unrelated SDR from Methylobacterium sp. 77 (M77_SDR) with distinct keto-reducing activity was engineered into a promiscuous enzyme with imine-reducing activity, thereby confirming that the ability to reduce imines can be rationally introduced into members of the “classical” SDR enzyme family. Thus, members of the SDR family could be a promising starting point for protein approaches to generate new imine-reducing enzymes.     

新鲜蛹虫草中SOD的提取工艺优化

以酶活力为考核指标,对新鲜蛹虫草中超氧化物歧化酶(SOD)的提取工艺进行了优化,通过单因素实验和L9(34)正交实验确定新鲜蛹虫草中SOD的最优提取条件为:新鲜蛹虫草用量0.5g、磷酸缓冲溶液用量20mL、磷酸缓冲溶液pH值8.0、超声提取时间20min、(NH4)2SO4饱和度42%,在此条件下,SOD酶活力最高。     

Angiotensin I‐converting enzyme inhibitory and Ca‐binding activities of peptides prepared from tuna cooking juice and spleen proteases

Tuna cooking juice is a by‐product from the tuna canning industry. In this study, tuna cooking juice was hydrolysed by proteases extracted from the spleen. Tuna cooking juice showed the highest ACE inhibitory and Ca‐binding activities after hydrolysis for 270 and 180 min, respectively. The hydrolysate was further fractionated by ultrafiltration. The permeate exhibited highest ACE inhibitory and Ca‐binding activities when passed through 1 and 5 kDa cut‐off membranes, respectively. Gel filtration chromatography was used to determine the MW of bioactive peptides that exhibited highest ACE inhibitory and Ca‐binding activities. Those peptides that exhibited highest ACE inhibitory and Ca‐binding activities were the MW range of 238–829 Da and 1355–1880 Da, respectively. These results suggest that the tuna cooking juice and the spleen protease extract are a potential source of bioactive peptides that can be utilised as bioactive ingredients in functional food and nutraceuticals.     

Flavourzyme酶水解蛋清蛋白的工艺研究

以水解度和蛋白质残留量为指标,研究温度、pH、酶用量及蛋清浓度对Flavourzyme酶解蛋清蛋白的影响,并通过正交试验来确定其最佳工艺。结果是温度55℃、pH6.5、加酶量为质量分数6%、蛋清料液质量体积比1 g:5 m L为最佳工艺参数。在该条件下,6 h酶解物的蛋白质残留率为27.68%,水解度为38.75%,水解物的相对分子质量大部分集中在300以下,即主要以游离氨基酸及二肽形式存在。