酶法提取金乌贼墨汁中黑色素的工艺条件研究

以金乌贼墨汁为原料,采用单因素试验,对胰蛋白酶,木瓜蛋白酶,胃蛋白酶,中性蛋白酶和碱性蛋白酶等五种酶提取黑色素的效果进行比较分析,筛选出水解效果较好的碱性蛋白酶。并通过正交试验,优化了碱性蛋白酶酶解金乌贼墨汁的最佳工艺,实验结果表明,在底物浓度2%、水解温度50℃、pH值为7.4,加酶量4200U/g和水解8h的条件下进行水解的效果较好。     

海马总蛋白提取及其酶解条件优化

为了提取海马总蛋白并确定最佳的酶解条件,将海马粉碎后采用水提法提取海马总蛋白。分别选用木瓜蛋白酶、碱性蛋白酶、中性蛋白酶和胰蛋白酶,在不同酶解p H、时间、温度和E/S条件下进行单因素和正交试验法对海马总蛋白的酶解条件进行优化。利用聚丙烯酰胺凝胶电泳和BCA法来分别检测不同条件下的酶解情况及其蛋白含量,根据蛋白水解度来确定其最佳的酶解条件。实验结果表明:从10 g干海马中提取总蛋白为1.056 g,蛋白浓度为0.106 g/m L。海马总蛋白的最佳水解酶为碱性蛋白酶,其最佳酶解条件为:p H 9,E/S为5%,温度为50℃,时间为3 h,获得海马总蛋白的最大水解度为96.9%。研究获得了海马总蛋白及其最佳酶解条件,并为海马多肽生物活性研究奠定了基础。     

海参制备胶原蛋白肽工艺研究

主要研究以海参体壁为原料制备胶原蛋白肽的工艺条件,通过单因素和正交试验对酶解条件进行优化。结果表明,使用木瓜蛋白酶的酶解效果最好,最佳酶解条件为:酶解温度55℃、p H值7.0、加酶量2%、料液比1:30和提取时间3 h,此条件下海参胶原蛋白肽水解度达到14.23%。     

双酶法制备螺旋藻多肽的工艺研究

选用碱性蛋白酶和木瓜蛋白酶结合的双酶法对螺旋藻蛋白进行水解。其中,对木瓜蛋白酶水解螺旋藻蛋白的工艺进行优化。以水解度为指标,研究了酶解时间、酶与底物比、pH和酶解温度4种因素对酶解反应的影响。在此基础上设计了3因素(加酶量、酶解温度和pH)3水平的响应面试验。结果表明碱性蛋白酶水解螺旋藻蛋白的最佳酶解条件为:加酶量4300 U/g,pH 7.0,酶解温度55℃,酶解时间160 min;木瓜蛋白酶的最佳酶解条件为:酶底比为4.5%,酶解温度60℃,pH 6.5,酶解时间210 min。利用碱性蛋白酶和木瓜蛋白酶结合的双酶法制得的多肽水解度可达32.90%,与单酶法相比,水解度明显提高。     

响应面法优化蚕豆蛋白酶解工艺条件的研究

考察了碱性蛋白酶、胰蛋白酶和中性蛋白酶对蚕豆蛋白的酶解效果,探讨了水解度（DH）与酶解产物抗氧化活性间的关系。通过单因素试验和响应面分析法,得到碱性蛋白酶酶解工艺的最佳条件。结果表明,温度50℃、pH8.0、酶底比8%、底物浓度3%条件下酶解3h,水解度0~22%内,碱性蛋白酶较胰蛋白酶和中性蛋白酶水解蚕豆蛋白效果好;DH与还原能力（R2=0.68~0.81）及ABTS清除能力（R2=0.98~0.99）具有较好的相关性,碱性蛋白酶酶解液较其他2个酶解液有较好的还原能力和ABTS清除能力;优化后的最佳酶解工艺参数为：酶底比8%,温度50℃、pH 7.6,对蚕豆蛋白还原能力的影响顺序为酶底比＞pH＞温度;在此条件下,蚕豆蛋白酶解液的还原能力理论值为0.174,验证试验测得还原能力为0.173,与理论值接近。     

碱性蛋白酶水解大豆分离蛋白制备大豆多肽的工艺条件优化

以大豆分离蛋白为原料,地衣芽胞杆菌产碱性蛋白酶水解大豆分离蛋白,分别从酶浓度、pH值、反应温度和水解时间等因素来研究水解效果,在50℃,pH 10,酶浓度100 U/mL,水解2 h时水解效果最好,水解度达到31.45%。     

木瓜蛋白酶法提取荸荠皮多糖

本文以低温烘干的荸荠皮为原料,利用木瓜蛋白酶提取荸荠皮中的多糖。通过单因素实验和正交实验研究了p H、木瓜蛋白酶用量、液料比、酶解温度、酶解时间对多糖提取效果的影响。结果表明,最佳提取工艺条件为:p H 4,木瓜蛋白酶用量0.3%,液料比25∶1 m L/g,酶解温度50℃,酶解时间2 h。验证实验表明,多糖平均得率可达30.03%,远高于相同条件下水提法的得率21.23%,且重现性好。本实验为提高荸荠的综合价值提供了理论基础和实验方法。     

酶解魔芋飞粉制备高F值寡肽最佳工艺条件的研究

目的:为了得到高F值寡肽,需要对蛋白质进行水解。方法:采用酶法水解魔芋飞粉制备高F值寡肽。通过对蛋白质水解度的测定,确定了碱性蛋白酶和链霉蛋白酶的最佳酶解条件:碱性蛋白酶加酶量0.1%,底物浓度3.75%,pH9.0,水解温度45℃,时间5h;链霉蛋白酶加酶量0.03%,pH8.0,水解温度50℃,时间7h。酶解液再经过凝胶层析分离纯化后可用于制备高F值寡肽。     

花椒籽黑色素提取和脱蛋白技术研究

采用二次回归正交旋转组合试验优化了花椒籽黑色素的浸提工艺,并用蛋白酶酶解-Sevag法联用脱除花椒籽黑色素中的蛋白.结果表明:影响花椒籽黑色素提取效果的因素依次为温度>NaOH浓度>料液比,花椒籽黑色素提取的优化工艺参数为NaOH浓度为1.20 mol/L,料液比为1∶24.64,温度为70℃下提取2 h,共2次,所得花椒籽色素粗品为棕黑色无定型粉末,得率为6.1%,色价为201.62,蛋白质含量为44.83%~48.63%.碱性蛋白酶脱蛋白条件为温度50℃,pH 9,加酶量为粗色素溶液(浓度为1 mg/mL)体积的6%,再用Sevag法处理3次,蛋白质脱除率可达81.23%,花椒籽黑色素色价可提高1.5倍.     

木瓜蛋白酶酶解鱼鳞提取胶原蛋白的工艺研究

用木瓜蛋白酶水解鱼鳞提取胶原蛋白,对影响酶解过程的的主要因素(酶量、温度、底物浓度)分别作为单因素进行了实验。并通过正交实验得到鱼鳞胶原蛋白提取条件的优化组合,同时对提取物进行了分析检测。结果表明,木瓜蛋白酶对鱼鳞有较好的水解效果,酶用量宜采用4g/L,最佳温度为60℃,底物浓度宜选择20%。     

On the origin of the Hirudinea and the demise of the Oligochaeta

The phylogenetic relationships of the Clitellata were investigated with a data set of published and new complete 18S rRNA gene sequences of 51 species representing 41 families. Sequences were aligned on the basis of a secondary structure model and analysed with maximum parsimony and maximum likelihood. In contrast to the latter method, parsimony did not recover the monophyly of Clitellata. However, a close scrutiny of the data suggested a spurious attraction between some polychaetes and clitellates. As a rule, molecular trees are closely aligned with morphology-based phylogenies. Acanthobdellida and Euhirudinea were reconciled in their traditional Hirudinea clade and were included in the Oligochaeta with the Branchiobdellida via the Lumbriculidae as a possible link between the two assemblages. While the 18S gene yielded a meaningful historical signal for determining relationships within clitellates, the exact position of Hirudinea and Branchiobdellida within oligochaetes remained unresolved. The lack of phylogenetic signal is interpreted as evidence for a rapid radiation of these taxa. The placement of Clitellata within the Polychaeta remained unresolved. The biological reality of polytomies within annelids is suggested and supports the hypothesis of an extremely ancient radiation of polychaetes and emergence of clitellates.     

On the ontogeny of the haptor and the evolution of the Monogenea

Data on the ontogeny of the posterior haptor of monogeneans were obtained from more than 150 publications and summarised. These data were plotted into diagrams showing evolutionary capacity levels based on the theory of a progressive evolution of marginal hooks, anchors and other attachment components of the posterior haptor in the Monogenea (Malmberg, 1986). 5 + 5 unhinged marginal hooks are assumed to be the most primitive monogenean haptoral condition. Thus the diagrams were founded on a 5 + 5 unhinged marginal hook evolutionary capacity level, and the evolutionary capacity levels of anchors and other haptoral attachement components were arranged according to haptoral ontogenetical sequences. In the final plotting diagram data on hosts, type of spermatozoa, oncomiracidial ciliation, sensilla pattern and protonephridial systems were also included. In this way a number of correlations were revealed. Thus, for example, the number of 5 + 5 marginal hooks correlates with the most primitive monogenean type of spermatozoon and with few sensillae, many ciliated cells and a simple protonephridial system in the oncomiracidium. On the basis of the reviewed data it is concluded that the ancient monogeneans with 5 + 5 unhinged marginal hooks were divided into two main lines, one retaining unhinged marginal hooks and the other evolving hinged marginal hooks. Both main lines have recent representatives at different marginal hook evolutionary capacity levels, i.e. monogeneans retaining a haptor with only marginal hooks. For the main line with hinged marginal hooks the name Articulon-choinea n. subclass is proposed. Members with 8 + 8 hinged marginal hooks only are here called Proanchorea n. superord. Monogeneans with unhinged marginal hooks only are here called Ananchorea n. superord. and three new families are erected for its recent members: Anonchohapteridae n. fam., Acolpentronidae n. fam. and Anacanthoridae n. fam. (with 7 + 7, 8 + 8 and 9 + 9 unhinged marginal hooks, respectively). Except for the families of Articulonchoinea (e.g. Acanthocotylidae, Gyrodactylidae, Tetraonchoididae) Bychowsky's (1957) division of the Monogenea into the Oligonchoinea and Polyonchoinea fits the proposed scheme, i.e. monogeneans with unhinged marginal hooks form one old group, the Oligonchoinea, which have 5 + 5 unhinged marginal hooks, and the other group form the Polyonchoinea, which (with the exception of the Hexabothriidae) has a greater number (7 + 7, 8 + 8 or 9 + 9) of unhinged marginal hooks. It is proposed that both these names, Oligonchoinea (sensu mihi) and Polyonchoinea (sensu mihi), will be retained on one side and Articulonchoinea placed on the other side, which reflects the early monogenean evolution. Except for the members of Ananchorea [Polyonchoinea], all members of the Oligonchoinea and Polyonchoinea have anchors, which imply that they are further evolved, i.e. have passed the 5 + 5 marginal hook evolutionary capacity level (Malmberg, 1986). There are two main types of anchors in the Monogenea: haptoral anchors, with anlages appearing in the haptor, and peduncular anchors, with anlages in the peduncle. There are two types of haptoral anchors: peripheral haptoral anchors, ontogenetically the oldest, and central haptoral anchors. Peduncular anchors, in turn, are ontogenetically younger than peripheral haptoral anchors. There may be two pairs of peduncular anchors: medial peduncular anchors, ontogentically the oldest, and lateral peduncular anchors. Only peduncular (not haptoral) anchors have anchor bars. Monogeneans with haptoral anchors are here called Mediohaptanchorea n. superord. and Laterohaptanchorea n. superord. or haptanchoreans. All oligonchoineans and the oldest polyonchoineans are haptanchoreans. Certain members of Calceostomatidae [Polyonchoinea] are the only monogeneans with both (peripheral) haptoral and peduncular anchors (one pair). These monogeneans are here called Mixanchorea n. superord. Polyonchoineans with peduncular anchors and unhinged marginal hooks are here called the Pedunculanchorea n. superord. The most primitive pedunculanchoreans have only one pair of peduncular anchors with an anchor bar, while the most advanced have both medial and lateral peduncular anchors; each pair having an anchor bar. Certain families of the Articulonchoinea, the Anchorea n. superord., also have peduncular anchors (parallel evolution): only one family, the Sundanonchidae n. fam., has both medial and lateral peduncular anchors, each anchor pair with an anchor bar. Evolutionary lines from different monogenean evolutionary capacity levels are discussed and a new system of classification for the Monogenea is proposed.In agreeing to publish this article, I recognise that its contents are controversial and contrary to generally accepted views on monogenean systematics and evolution. I have anticipated a reaction to the article by inviting senior workers in the field to comment upon it: their views will be reported in a future issue of this journal. EditorIn agreeing to publish this article, I recognise that its contents are controversial and contrary to generally accepted views on monogenean systematics and evolution. I have anticipated a reaction to the article by inviting senior workers in the field to comment upon it: their views will be reported in a future issue of this journal. Editor