Phytase Producing Marine Yeasts: Diversity, Identification and Purification of Phytase from Hansenia

来源 :中国海洋大学 | 被引量 : 0次 | 上传用户:bravehearterdoctor
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
Marine yeast strains isolated from various marine habitats were studied from 2003 November to 2005 November. Isolation of yeasts strains were carried out under sterile conditions and were stored at -70°C at the laboratory of Molecular microbiology, UNESCO Chinese Center of Marine Biotechnology, Ocean University of China, Yushan Road, No. 5, China.During the experimental period, about 300 strains were isolated. Among them, a total of 177 isolates were classified to species level and 29 morphologically distinct yeast species were identified and genotyped. The highest species diversity was observed in sea water samples. Yarrowia lipolytica (39.8%) is the most common species identified yet host specific or habitat specific relationship was not clearly observed in this study. The complete YPD agar medium was used to store the yeast strains.Isolated strains were then screened for phytase activity using a modified minimal synthetic medium, which contained vitamins, minerals, salts, ammonium sulfate, agar and 0.5% sodium phytate as a sole source of carbon and phosphate. Strains with significant growth were selected and subjected for further enzyme assay. As a result ten strains exhibited comparatively high enzyme activity and Yarrowia lipolytica (W2B) and Kodamea ohmeri (BG3) had the highest extra-cellular phytase activity. Strain YF 12C indicated the highest cell bound enzyme activity and this strain was screened from the gut of Sea cucumber (Holothuria scabra). Interestingly, the optimum pH and temperature for crude enzyme of all strains were between 5 - 9 and 50°C– 65°C respectively.Among these ten isolates, WZ1 was from the gut of fish (Scomberomorus niphonius), W2B and YF12C from sea cucumber (Holothuria scabra), whereas BG3, YF04C and YF08 were from the gut of fish species (Hexagrammes otakii and Synecogobius hasts). The sea water isolates were N12C and MA6 including NY4E and MB2 from Salterns and Southern sea of China respectively. The results of this study revealed that the predominant phytase secreting strains were the ones isolated from the gut of marine animals. In addition, this study indicated the diverse habitats of these strains. To date, few terrestrial yeasts species have been reported as phytase producers but marine yeasts were in scarce as we used in this research.Ten strains with relatively high phytase activity were then characterized by genetic, physiological and biochemical analyses to clarify their taxonomic position. The 18S rDNA partial genomic sequences and ITS sequences of the yeast strains were examined and analyzed for similarity comparison using NCBI BLAST program and deposited at NCBI data bank. Phylogenetic tree was constructed by using PHILIP program. Taxonomic studies confirmed that strains WZ1, W2B, BG3, YF04C, and YF08 were similar to Hanseniaspora uvarum, Yarrowia lipolytica, Kodamaea ohmeri, Issatchenkia orientalis and Yarrowia lipolytica, respectively, while both N12C and NY4E to Candida carpophila, and MA6, YF12C and MB2 were similar to Candida tropicalis, respectively.In the current study, yeast strain Hanseniaspora uvarum - WZ1 was selected for purification and was grown for four days at 28°C in optimized medium. Extra-cellular phytase from was purified using gel filtration chromatography column of Sephadex TM G-100. The optimum pH and temperature of the purified enzyme was pH 5 and 60°C, respectively. These values were different from that of the crude enzyme. The molecular weight was about 46.3 kDa. Purified phytase was not inhibited by CaCl2, MgCl2, NH4Cl and ZnCl2 suggesting that Ca+2, Mg+2 NH4+ and Zn+2 ions are not essential for activity. The enzyme was strongly inhibited by Cu +2 however, Co +, Li + and Mn +2 were less effective. The Km and V max values were 0.392 mmol and 3.889 mmol/min.
其他文献
[背景]骨髓增生异常综合征(myelodysplastic syndromes,MDS)常有多个基因突变,且基因突变影响患者生存预后。SF3B1突变的MDS患者骨髓环状铁粒幼红细胞(ringsideroblasts,RS)比例增高,总体生存预后较好。WHO(2016)诊断标准建议SF3B1突变但不伴有原始细胞增多(<5%)的MDS患者仅需骨髓RS≥5%即可诊断为伴环状铁粒幼红细胞增多的骨髓增生异常
低摩擦技术是内燃机的变革性技术之一,降低内燃机摩擦损失最简单且最经济的方式是直接减小润滑油粘度,但低粘度油的应用将导致磨损加剧等问题,因此开展应用低粘度油的多缸内燃机主轴承系润滑-磨损动态耦合仿真研究十分必要。本文针对低摩擦多缸内燃机主轴承局部总压和剪切率升高的工作特点,提出了综合考虑粘压效应和剪切稀化效应的润滑剂非线性粘度模型,实现了面向低粘度油应用的混合润滑条件下多缸内燃机主轴承系瞬态润滑仿真
代数图论是图论的重要组成部分.图的自同态半群的结构问题因深刻揭示图论和半群理论之间的关系而成为代数图论的重要研究内容C.Godsil等人在代数图论的经典教材的第六章中详细地介绍了图的自同态半群的结构与图的结构(特别是图的色数)之间的紧密联系M.Petrich和N.R.Reilly指出:正则半群(即半群中每个元素都有所谓的伪逆)由于其与群最为相像,因而在半群理论中占有重要地位.如果图的自同态半群是正
根据形态上的差异细胞死亡可以分为细胞凋亡、细胞自噬和细胞坏死。最初细胞坏死被认为是一种由化学因素、物理因素以及病原体感染造成的被动细胞死亡。但是越来越多的证据表明,细胞坏死也可以是一个主动的、受到严格调控的过程,即程序性细胞坏死。程序性细胞坏死参与一系列生理以及病理活动,对于维持生物体内稳态非常重要。目前了解较为深入的程序性细胞坏死主要有肿瘤坏死因子(tumor necrosis factor,
在秩为1的格的顶点算子代数中,对应于Jack函数,我们定义了一个顶点算子。我们把这个顶点算子称为Jack顶点算子。我们给出一个一般公式把Jack顶点算子积表为广义的齐次对称函数的组合。对Jack顶点算子,我们导出一个Jacobi-Trudi型公式,从而表明在某些条件下顶点算子积这个集合是表示空间的一个基。作为应用,我们实现了形状为矩形的Jack函数和形状为去除右下角的那个块的矩形的Jack函数。进
背景 凝血异常和纤维蛋白沉积见于人类许多类型的炎症反应,并且与炎症损害密切相关,基础研究表明,凝血系统参与炎症调节。临床资料和我们的前期研究发现,老年机体存在高凝状态,同时老年个体较青年个体对各种炎症刺激和病理损伤更加易感,因此我们推测老年个体的高凝状态促进了炎症反应。但老年个体高凝状态的原因及其对老年个体炎症反应影响的机制目前尚不十分清楚。目的 为验证我们的假说,本研究首先观察生理与炎症状态下老
气固流化床干法分选技术为世界干旱地区煤炭提供了一种高效分选方法。在工业性分选试验过程中,该技术所用的加重质(磁铁矿粉)粒级较窄,存在制备困难,成本较高的问题。此外,鉴于传统选煤厂建筑方式基建投资较大,中国矿业大学与唐山神州机械有限公司合作建立了40~60t/h的KZX40型模块式气固流化床干法选煤系统。因此,有必要拓展加重质适用粒级,增强该技术的适用性。本文应用实验与数值计算相结合的方法,研究了不
This thesis mainly discusses Dirichlet spaces of one-dimensional diffusions and their regular subspaces. We show that the ε1-closure of an algebra of bounded functions in a Dirichlet space has Markovi
学位
古人云,不以规矩,不成方圆。可见凡事都有“规矩”。翻译活动亦如此,翻译标准即是支配翻译活动运行的“规矩”之一。翻译标准在中西翻译史上都发挥了重要作用,不仅在实践中规范译者的翻译行为,也在理论研究中成为了一个重要议题。尽管中西方对翻译标准有不同的认识,译者在翻译实践中依据的翻译标准也不尽相同,翻译理论家厘定的具体翻译标准也各式各样,“翻译标准”这一概念所用的术语也有差异,但不可否认翻译标准的确客观存
本文主要研究组合优化中若干问题的近似算法。文章首先介绍了组合优化的概貌,给出了近似算法定义及其性能度量标准。在第二章至第五章中,我们分别研究了四个组合优化问题的近似算法,并给出了它们的性能分析。 第二章研究了以极大化最小负载为目标的3—分划问题和带核3—分划问题,我们证明了对3—分划问题,MLPT算法的最坏情况界为(3m-1)/(4m-2),对带核3—分划问题,MLPT算法的最坏情况界(2m