A 160-kDa Protein Is Essential for Hemocyanin-derived Melanosis of Prawn

ABSTRACT A purified 160-kDa protein, isolated from the cuticle of kurama prawn and named melanosis collaborating factor (MCF), was a key factor in promoting melanosis by a cooperative reaction with hemocyanin-derived phenoloxidase but hemocyanin itself was incapable of producing black pigment. The enzymatic activity of MCF in the body of the prawn was very stable against the process of freezing and thawing; the enzyme maintained more than 80% of its activity after 3 mo of storage at −2 5 °C. These results indicated that MCF, acting in conjunction with hemocyanin-derived phenoloxidase, played a crucial role in postharvest melanogenesis in prawn.     

虾类保藏过程中酚氧化酶酶促黑变作用机理及其抑制方法的研究进展

在虾类贮藏过程中,虾体的黑变严重影响了虾的感官品质,酚氧化酶是催化酪氨酸等色原物质转化为黑色素造成虾体黑变的主要因素。文章综述了国内外酚氧化酶的形成机制研究现状,重点叙述了虾类中酚氧化酶的特性及抑制虾类黑变方法的研究进展,为虾类贮藏保鲜技术的研发与改进提供参考。     

曲酸对南美白对虾酚氧化酶活性和结构的影响

为研究曲酸对南美白对虾酚氧化酶(phenol oxidase,PO)活性和结构的影响,以南美白对虾头胸部分离纯化出的PO为原料,研究不同浓度曲酸(0、0.1、0.3、0.5、0.7、0.9、1.1、1.3、1.5 mmol/L)对PO活性的抑制,确定曲酸作用PO的抑制类型;通过测定不同浓度曲酸(0、0.1、0.3、0.5、0.7、0.9、1.1 mmol/L)处理的PO蛋白内源荧光光谱、表面疏水性和二级结构类型含量的变化,研究曲酸对PO空间结构的影响。结果表明:随着作用PO曲酸浓度的增加,PO的剩余酶活逐渐减少,当曲酸浓度达到1.5 mmol/L时,PO的剩余酶活仅为9.8%,参与的反应体系褐变指数变化值为3.02,抑褐变效果明显;曲酸能够有效的抑制PO活性,曲酸作用PO的抑制类型属于竞争性抑制剂;随着作用PO曲酸浓度的增加,PO的内源荧光强度不断降低,最大发射波长发生红移,PO蛋白表面疏水性先增加后降低;随着作用PO曲酸浓度的增加,二级结构类型α-螺旋和β-折叠含量降低,无规则卷曲和β-转角增加。可以推测,曲酸与PO结合后,酶蛋白空间结构发生了改变。     

Activation Mechanisms of Pro-Phenoloxidase on Melanosis Development in Florida Spiny Lobster (Panulirus argus) Cuticle

Phenoloxidase (PO), pro-PO [1] and pro-PO [2], isolated from cuticular extracts of early premolt spiny lobster, resolved into doublets of molecular weights 156, 151, and 96.0, 93 kD, respectively. Trypsin-activated forms of pro-PO [1] and pro-PO [2] were 123 and 83.6 kD, respectively. Apparently melanosis of shell and hyperdermal tissue in lobsters was related to stage of molt. The pro-PO forms were activated in vitro by extracts of molting fluid which is secreted into the cuticle just before molt, and the activated PO co-migrated in SDS-PAGE with the endogenously activated PO. The molting fluid was hypothesized to be the source of the natural activator(s) of pro-PO.     

57个昆虫酚氧化酶抑制剂的QSAR研究

使用分子电性距离矢量（MEDV-13）描述子和顶点度-距离指数（VDI）描述子提取57个酚氧化酶抑制剂分子结构信息,基于预测的VSMP方法筛选最佳描述子;用多元线性回归法构建了一个8变量QSAR模型,该模型的估计相关系数R2=0.969 8,LOO交互检验相关系数R2cv=0.960 6,显示出模型具有良好的稳定性和预测能力。计算了最佳描述子8个变量的碎片贡献率。研究结果表明,昆虫酚氧化酶抑制剂的生物活性IC50主要受苯环、C原子、N原子、O原子、S原子以及分子中原子间连接情况的影响。     

南美白对虾酚氧化酶的提取及生化特性

以南美白对虾为研究对象,分析以Tris-HCl为提取缓冲液,研究提取液浓度、提取液pH值、浸提时间对酚氧化酶活力的影响,以L9（33）正交试验方法优化酚氧化酶提取条件,然后对南美白对虾酚氧化酶生化特性进行研究。结果表明：提取液浓度为0.2 mol/L、提取液pH值为8.0、浸提时间为20 min条件下提取的酶具有最大的活力;以邻苯二酚为底物时,酚氧化酶最适pH值为8、最适温度为40℃,在该条件下酶促反应动力学符合米氏方程,米氏常数Km为0.717 mol/L,Vmax为0.130 A/min。     

三疣梭子蟹酚氧化酶的生化特性研究

对三疣梭子蟹酚氧化酶的生化特性进行研究。结果表明：以邻苯二酚为底物,酚氧化酶在pH7.8～8.4时最稳定,最适pH 值为8.2。温度55℃时,酚氧化酶活力最大,在25～40℃时,热稳定性最好。酶促褐变反应动力学符合米氏方程,温度为30℃,pH7.2 时,米氏常数Km=0.531mol/L,最大反应速率0.049 A530nm/min。选用的12 种抑制剂对三疣梭子蟹酚氧化酶均有一定的抑制效果,以植酸抑制效果最强,焦亚硫酸钠、L- 半胱氨酸、草酸次之,抗坏血酸效果较好。     

取代水杨醛席夫碱的合成及其对昆虫酚氧化酶的抑制活性

由取代水杨醛与取代苯胺反应,合成了两类14种取代水杨醛席夫碱化合物,其结构经红外光谱、核磁共振氢谱及元素分析确证。测试了目标化合物对菜青虫酚氧化酶活力的抑制作用,结果表明,在浓度为0.1 mmol/L和0.2 mmol/L时部分化合物对该酶有较好的抑制活性。     

南美白对虾血细胞酚氧化酶活力的研究

研究了凡纳滨对虾(Penaeus vannamei)血细胞酚氧化酶在不同温度、pH及在一定浓度的Ca2 和SDS下酶活力的变化情况.试验结果表明:在自然状态下,南美白对虾血淋巴中的血细胞内存在酚氧化酶,但更多的是以酶原的形式存在于血细胞内,不显活性;Ca2 、SDS能激活酚氧化酶原,提高PO活力;在一定范围内改变pH值,或提高温度也能够提高酶活力.该酶的最适pH值为6.0,最适作用温度为50℃.     

Degradation of Phenolic Compounds in Coal Gasification Wastewater by Biofilm Reactor with Isolated Klebsiella sp.

This study was conducted to evaluate the degradation of phenolic compounds by one strain isolated from coal gasification wastewater（ CGW）. 16S rRNA gene sequences homology and phylogenetic analysis showed that the isolate is belonged to the genus Klebsiella sp. The effect of different phenolic compounds on the isolate was investigated by determining OD600and phenoloxidase activity,of which the results showed that the isolate can utilize phenol,4-methyl phenol,3,5-dimethyl phenol and resorcinol as carbon resources. The biofilm reactor（ formed by the isolate） can resist the influent concentration of phenolic compounds as high as750 mg /L when fed with synthetic CGW and incubated at optimum conditions. The capacity of improving the biodegradability of CGW through degrading phenolic compounds was testified with fed the biofilm reactor with real CGW. Thus,it might be an effective strain for bioaugmentation of CGW treatment.