80 resultados para Kappa-casein
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In order to study the relationship between chemical structure and properties of modified carrageenans versus antioxidant activity in vitro, K-carrageenan oligosaccharides were prepared through mild hydrochloric acid hydrolysis of the polysaccharide, and these were used as starting materials for the partial synthesis of their oversulfated, acetylated, and phosphorylated derivatives. The structure and substitution pattern of the oligosaccharides and their derivatives were Studied using FTIR and C-13 NMR spectroscopy, and their in vitro antioxidant activities were investigated. Certain derivatives of the carrageenan oligosaccharides exhibited higher antioxidant activity than the polysaccharides and oligosaccharides in certain antioxidant systems. The oversulfated and acetylated derivatives, which scavenge superoxide radicals, the phosphorylated and low-DS acetylated derivatives, which scavenge hydroxyl radicals, and the phosphorylated derivatives, which scavenge DPPH radicals, all exhibited significant antioxidant activities it, the systems examined. The effect of the molecular weight of the carrageenan on antioxidant activities, however, is not obvious from these studies. (c) 2005 Elsevier Ltd. All rights reserved.
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Oligosaccharides were prepared through mild hydrochloric acid hydrolysis of kappa-carrageenan from Kappaphycus striatum to compare the antitumor activity with carrageenan polysaccharides. Oligosaccharide fractions were isolated by gel permeation chromatography and the structure of fraction 1 (F1) was studied by using negative- ion electrospray ionization-mass spectrometry (ESI-MS), and H-1 and C-13-NMR spectrometry. The in vitro antitumor effects in three human neoplastic cell lines (KB, BGC, and Hela) of polysaccharides and F1 were investigated. The bioassay results showed that F1 exhibited relatively higher antitumor activity against the three cancer cells than polysaccharides.
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Antioxidant activity of kappa-carrageenan oligosaccharides (OM) and their chemical modification derivatives was investigated employing various established in vitro systems, such as reducing power, iron ion chelation, and total antioxidant activity using beta-carotene-linoleic acid system. The oversulfated (SD), lowly (LAD), and highly acetylated derivatives (HAD) in reducing power assay, the phosphorylated derivative (PD) in metal chelating assay, and oversulfated and phosphorylated derivatives in total antioxidant activity assay exhibited antioxidant activity higher than that of carrageenan oligosaccharides. The results indicated that the chemical modification of carrageenan oligosaccharides can enhance their antioxidant activity in vitro. The protective effects of the carrageenan oligosaccharides and their chemically modified derivatives against H2O2 and UVA (long-wave ultraviolet radiation) induced oxidative damage on rat thymic lymphocyte were investigated by measuring cell viability via 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT). Thymic lymphocyte exposure to H2O2 and UVA, a marked reduction in cell survival was observed, which was significantly prevented by carrageenan oligosaccharides and their derivatives (preincubated for 2 h) at 66.7-2000 mu g/mL. But both the carrageenan oligosaccharides and their different derivatives showed the similar protective effects on intracellular level. Taken together, these results suggest that carrageenan oligosaccharides and their derivatives show relevant antioxidant activity both in vitro and in a cell system. (C) 2005 Elsevier Ltd. All rights reserved.
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The modulation of carrageenan oligosaccharides from Kappaphycus striatum on the immune system in S 180-bearing mice was investigated. The mice inoculated with S180 cell suspension were treated p.o. with carrageenan oligosaccharides (50, 100 and 200 mu g/g) for 14 days. The effects of carrageenan oligosaccharides on transplantable tumors and macrophage phagocytosis, quantitative hemolysis of sheep red blood cells (QHS),. lymphocyte proliferation, the activity of natural killer cells (NK), production of interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha) were studied. Carrageenan oligosaccharides could significantly inhibit the growth of transplantable sarcoma S180 and increase macrophage phagocytosis, the form of antibody secreted by spleen cells, spleen lymphocyte proliferation, NK cells activity, serumal IL-2 and TNF-alpha level in S 180-bearing mice. Considering all these results, it is suggested that carrageenan oligosaccharides exert their antitumor effect by promoting the immune system. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
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Chemical examination of the green alga Cladophora fascicularis resulted in the isolation and characterization of a new porphyrin derivative, porphyrinolactone (1), along with five known phaeophytins 2-6 and fourteen sterols and cycloartanes. The structure of 1 was determined on the basis of spectroscopic analyses and by comparison of its NMR data with those of known phaeophytins. Compounds 1-6 displayed moderate inhibition of tumor necrosis factor alpha (TNF-alpha) induced nuclear factor-kappa B (NF-kappa B) activation, while 2 and 4 displayed potential inhibitory activity toward proteasome chymotripsin-like activation. The primary structure-activity relationship was also discussed.
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In the title compound, [CdCl2(C13H12N2O)(2)], the Cd-II ion is situated on an inversion centre, coordinated by two 0 atoms [Cd-O=2.3878 (17) angstrom] and two N atoms [Cd-N = 2.3404 (15) angstrom] from two N-(2-pyridylmethyl)benzamide ligands, and two Cl atoms [Cd-Cl = 2.5566 (6) angstrom], in a distorted octahedral geometry. In the crystal structure, intermolecular N-H center dot center dot center dot Cl hydrogen bonds [N center dot center dot center dot Cl = 3.1705 (18) angstrom] and pi-pi interactions, with a distance of 3.868 (3) angstrom between the centroids of the phenyl and pyridyl rings of neighbouring molecules, lead to the formation of two-dimensional layers parallel to the bc plane.
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The title compound, [ CdCl2( C12H8N2)(2)]center dot 0.5H(2)O, crystallizes with two independent complex molecules and one water molecule in the asymmetric unit. The Cd atoms in both independent complexes display a distorted octahedral coordination geometry formed by four N atoms from two phenanthroline ligands and two Cl atoms. In the crystal structure, pi-pi stacking interactions link complexes in two symmetry- independent ladders parallel to the c axis. Intermolecular O-H center dot center dot center dot Cl hydrogen bonds stabilize the crystal packing.
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In the title compound, [Cu(NCS)(2)(C5H6N2)(2)], each Cu atom is coordinated by two N atoms from two Eim (Eim = 1-vinyl-1H-imidazole) ligands and two N atoms from two isothiocyanate groups. The Cu atom adopts a square-planar geometry. The mean Cu-N(Eim) and Cu-N(NCS) distances are 1.960 (6) and 1.993 (6) angstrom, respectively.
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(第二部分的摘要) 酪蛋白激酶在许多物种的细胞分裂及分化过程中都有重要作用。在水稻中,以经过油菜素内酯处理的水稻幼苗为材料,通过cDNA微矩阵的方法得到了一个全长1939bp的基因OsCKI1(Accession number AJ487966)。该基因编码的蛋白产物属I型酪蛋白激酶(CKIs),含463个氨基酸。RT-PCR及Northern blot结果显示,基因OsCKI1在水稻各组织中表现为组成型表达,并且其表达受油菜素内酯(BR)及脱落酸(ABA)的诱导。在大肠杆菌中对该基因进行原核表达,并用表达后的蛋白粗提物进行酶活测定,显示该蛋白产物可磷酸化CKIs的特异性底物酪蛋白。通过构建OsCKI1的反义载体并转化水稻,对该基因的生理功能进行了研究。对转基因植株的纯系表型进行了观察,显示其根部发育异常,表现为具有较短的初生根、侧根及不定根数目少于对照。进一步研究显示初生根的变短是由于细胞延伸受抑制引起的。以CKI的特异性抑制剂,CKI-7处理野生型植株,也对OsCKI1缺失引起的表型进行了确认。值得注意的是,以外源生长素(IAA)处理转基因及经CKI-7处理过的野生型植株,都能恢复根部表型,使其生长正常。对反义植株初生根及次生根的游离生长素含量测定结果显示,OsCKI1可能在IAA的代谢途径中发挥作用。转基因植株的种子在萌发时对ABA及BR的处理都表现为不敏感,暗示该基因可能在各种激素信号转导途径中都有作用。OsCKI1-GFP双元表达载体的亚细胞定位的研究显示该基因主要定位于核中,可能参与了基因表达的调节。同时,以该反义转基因植株为材料,通过cDNA芯片的技术研究了受OsCKI1调节的基因的表达谱,结果显示该基因的缺失的确影响了参与信号转导及激素代谢途径的许多基因的表达。 (第四部分的摘要) 以OsCKI1反义转基因植株对照植株为材料,研究它们处于4℃低温胁迫下的反应情况。植株种子在室温下萌发并生长一段时间后,移入4℃低温下进一步生长。取对照及低温处理后的材料,对其表型进行观察,显示低温下转基因植株初生根生长受抑制程度小于对照,其生长的延缓程度低;相对电导率测定结果显示,经低温处理后,转基因植株相对电导率变化较小,质膜受害程度小;微管观察结果也显示在短期低温处理下对照根部延伸区细胞的皮层微管解聚,而转基因植株其根部延伸区细胞的皮层微管仍能保持正常状态。基因OsCKI1在低温下的表达模式表现为先升高之后又降低,推测其在低温信号的转导途径中发挥作用。通过总结以上结果,我们认为基因OsCKI1的反义转基因植株虽然在短期冷害下具有一定的抗冷能力,但其不具备形成长期稳定的冷适应的能力。
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本研究应用数字地球技术,基于1950年~1980年的全国958个气象站的基本气象数据(包括气象站的经纬度,海拔,月均温,月降水,年均温, 年降水,日照时数,日照百分率,风速等),比较了四个不同的根据水、热平衡原理设计的气候一植被关系模型(Penman模型、Holdridge生命地带系统、和Kira方法Thomthwaite模型)在中国应用的一致性和适用性。结果表明:(1) Penman模型在温带草原区和青藏高原地区的一致性指数超过50%,在青藏高原最出色,最有发展潜力。(2) Thornthwaite模型在热带雨林、季雨林区达到39. 72%,可以弥补Holdridg模型在热带地区分类精度的不足。(3) Holdridg生命地带系统在不同地带间适用性最广;只在热带地区,例如西部季雨林、雨林区域(52)、西部草原亚区域(63)和青藏高原温性荒漠地带(86)以及青藏高原温性草原地带(84)不理想。(4)吉良(Kira)方法在亚热带常绿阔叶林区可与Holdridg模型相媲美;在低海拔和湿润、半湿润地区效果尚可,但在温带荒漠区与青藏高原区的模拟效果与实际相差较远。 这四个传统的分类方法在中国植被区划一级分类上是适用的,Holdridge生命地带系统KAPPA -致性指数达到0.57模拟效果优于其它三者,但在特定地区,如青藏高原,所有模型均需改进优化或启用新的模型因子才能很好地区分植被亚地带。本研究还指出,数字地球技术的应用有助于推动气候一植被关系的研究,尤其在气候一植被指标(气候参数和模型参数)的大范围实时动态监测、气候一植被关系数据的海量信息高效、有序基础管理和功能型模型库支撑框架体系方面。
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神农架位于湖北省西部,长江以北、汉水以南的广阔地带,属北亚热带向暖温带的过渡区域。本文依据该地区所处地理位置的植被分布规律等资料,绘制了1:20万的植被复原图。并在此基础上,运用ERDAS imagine 8.4和Maplnfoprofessional 6.0软件,分别对神农架地区的TM影像(5、4、3波段)进行监督分类及目视解译,同时结合野外的样方调查,绘制了神农架地区1:20万的植被类型图,并建立了相应的属性数据库。最后,根据野外的GPS定位点对制图精度进行了Kappa检验。 制图结果表明,制图区总面积3476.67 km2,共计504个斑块。据统计,林地面积2607.45 km2,森林覆盖率75%;山地灌丛及亚高山灌丛总面积358.62 km2,占总面积的10.3%:草甸面积156.84 km2,占4.51%。自然植被划分为10个植被型,46个群系,以及农田(包括居民点)和茶园两种农业土地利用类型。其中针叶、落叶阔叶混交林面积最大(6个群系),约908 km2,占总面积的26.12%;其它依次为落叶阔叶林(1 1个群系),针叶林(4个群系),常绿阔叶林(3个群系),山地灌丛(5个群系),常绿阔叶、落叶阔叶混交林(3个群系),亚高山灌丛(6个群系),草甸(4个群系)以及亚高山针叶林(3个群系)等。另外,两种农业土地利用类型面积共计430 kn12,占总面积的12.37%。 植被类型图与复原图叠加分析表明:①常绿阔叶林的理论分布区域,由常绿阔叶林,常绿、落叶阔叶混交林等7种植被型以及农田(含居民点)等土地利用类型共同组成。因处低海拔区域,人口集中,所以农田(含居民点)分布最广,所占面积最大,占到该区域面积的35.28%;加上长期的人为干扰,常绿阔叶林面积缩小至48.76 krr12,占到该区域面积的13.93%;②常绿、落叶阔叶混交林的理论分布区域内,因干扰后落叶阔叶林恢复较快,逐渐占据优势。另外,该区域海拔较低,人类活动也较频繁,农田(含居民点)面积仍有相当的比例;③针阔混交林理论分布区海拔位置高,人为活动影响少,原地带性植被保存较好,分布面积最大;其余部分为落阔林等7种植被型共同组成。④针叶林理论分布区域应是以巴山冷杉林为单优种的亚高山针叶林带,但因历史上的皆伐及火烧等原因,现面积仅有17 kfr12,占该区域的19.8%,其余则为亚高山灌丛及亚高山草甸所替代。
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紫茎泽兰(Eupatorium adenophorum Spreng.)作为重要的外来入侵种之一,已严重破坏了入侵地生态系统的结构与功能,同时使当地农林业生产蒙受巨大的损失。因此,明确紫茎泽兰的竞争策略与扩散潜力,对制定合理的控制措施具有重要意义。 土壤氮素增加常常有助于外来种的入侵,那么降氮能否抵制入侵呢?向土壤中添加碳素,增强微生物对氮素的吸收,从而降低土壤中可被植物吸收的氮素是目前常用的降氮方法。为检验这一假说对入侵种紫茎泽兰的适用性,选取入侵种紫茎泽兰和本地种禾本科雀麦(Bromus japonicus)、菊科鱼眼草(Dichrocephala integrifolia),分别单栽或与紫茎泽兰混栽,向各植物组合中添加碳素(按1650克有机质/平方米计)或氮素(按30克氮素/平方米计),并设立对照进行盆栽实验。在碳素添加条件下,紫茎泽兰地上与地下生物量分别比对照降低了71.9% 和 74.9%,而本地种的生物量却没有受到显著影响。同时,碳素添加导致紫茎泽兰对本地种的竞争强度(RII)明显下降。氮素添加则减弱或逆转了上述碳素的作用。因此,向土壤中添加碳素有助于抵抗氮沉降对外来种入侵的促进作用,特别对抵御那些与氮素有亲和力的入侵种更为有效。在不同氮素条件下,紫茎泽兰对本地种竞争中,资源竞争强度均明显大于化感作用。尽管在紫茎泽兰发育早期化感作用较小,但是,碳素添加使紫茎泽兰可利用氮素减少,激发了化感作用在竞争中占有更大的比例,说明资源竞争与化感是相互结合、共同发挥作用的。 去叶作为模拟采食方法之一,至少可以通过两种方式影响植物竞争。一是去叶对目标种生长产生负面影响,二是邻种去叶有利于目标种生长,进而改变种间竞争关系。基于以上假定,对紫茎泽兰与相邻本地种分别单栽、同种合栽和异种混栽,进行去叶处理。结果恰与假定相悖:在整个生长季内,去叶增加目标株生物量的1.0 – 198.9%。对于入侵种而言,当邻居本地种去叶时,紫茎泽兰能完全弥补竞争产生的负面效应,即它与未去叶的单栽紫茎泽兰生长无显著差异;而紫茎泽兰去叶降低了相邻本地种的生长,加剧了原本由于竞争给对本地种带来的负面影响。因此,去叶效应与邻种的存在与否及邻种的类型均有关。在应用生物控制对目标株采食时,特别是在没有造成严重伤害或致死情况下,可以使本来具有竞争优势的入侵种变得更具强势。竞争与去叶之间存在复杂的相互作用,采食带来的间接效应可能比预想的更为复杂。因此,在生物控制措施实施之前,充分地估计生物控制的间接作用至关重要。 为检验反应—扩散模型(reaction–diffusion model)能否正确模拟紫茎泽兰小尺度扩散速率,本研究调查了四川攀枝花山区5个不同的生境的紫茎泽兰群落。对各样地内紫茎泽兰定位和年龄确定,基于克里金插值法做出其入侵格局的等年线图,通过等年线之间的垂直距离即可得到紫茎泽兰扩散速率的经验值。同时,计算内禀增长率和扩散系数,根据模型计算得到扩散速率的预测值。结果表明:紫茎泽兰扩散速率经验值明显小于模型预测值。因此,尽管反应—扩散模型已在大尺度扩散预测方面得到广泛的应用和验证,但在小尺度上,由于没有借助于复杂的生活史模型和分层模型,单凭反应—扩散模型不足以得到准确的预测。另外,在没有遭受干扰的生境中,可以发现紫茎泽兰扩散早期存在明显的时滞期。倘若这种时滞效应在入侵扩散中是频繁发生的,那么在判定物种是否具有入侵性时,很可能由于物种处于时滞期而被误认为非入侵种。 基于紫茎泽兰入侵至中国60年来所收集到的441已知分布记录和23个环境变量,采用GARP模型对其潜在分布范围做出了预测,并通过Kappa和ROC(receiver–operator characteristic)对预测结果进行检验。结果表明紫茎泽兰目前分布以云贵高原为主体,逐渐向四川盆地东部、山东半岛、东南沿海、台湾沿海以及海南岛等地扩散。冷季降水量、年极端低温和年均最高温对分布影响较大。RDA(Redundancy Analysis)分析结果显示温度和降水是限制紫茎泽兰扩散的重要因素。在过去的20年里,紫茎泽兰已经从温度较高、天气波动较小的亚热带气候逐渐向气候温凉、降水减少的高海拔地区扩散。紫茎泽兰可以适应相对较宽的温度和降水梯度,年均温度在10—22 ˚C,年降水量在800—2000 mm区间均有分布。本研究结果为发展早期预警工作、防止紫茎泽兰在中国进一步入侵提供了科学依据。
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在我国,小叶锦鸡儿(Caragana microphylla)、中间锦鸡儿(Caragana intermedia)、柠条锦鸡儿(Caragana korshinskii)、藏锦鸡儿(Caragana tibetica)和狭叶锦鸡儿(Caragana stenophylla)主要分布于北方温带地区,是造林、固沙、饲用及观赏的重要灌木,而贝加尔针茅(Stipa baicalensisi)、大针茅(Stipa grandis)、克氏针茅(Stipa krylovii)、本氏针茅(Stipa bungeana)、短花针茅(Stipa breviflora)、沙生针茅(Stipa glareosa)和戈壁针茅(Stipa gobica)是我国北方温带地区具有重要饲用价值和水土保持功效的多年生密丛禾草。这5种锦鸡儿灌木和7种针茅草本植物在东北、华北和西北地区的生态环境建设及社会经济发展中发挥特殊作用。关于它们地理分布与气候关系的深入研究十分必要,可以为其种质资源的开发和改善生态环境提供理论依据。 本研究首先全面收集这12个物种在中国北方温带干旱-半干旱地区的全部地理分布资料,利用ArcGIS 9.0软件绘制现状分布图。通过分析其现实分布格局,发现小叶锦鸡儿、中间锦鸡儿和柠条锦鸡儿在空间上呈现出从东到西的地理替代分布格局,继续向西南方向则分布有藏锦鸡儿,向西北方向分布有狭叶锦鸡儿。贝加尔针茅、大针茅、克氏针茅和戈壁针茅也在空间上呈现出自东向西的地理替代分布格局,克氏针茅向南被本氏针茅替代,短花针茅和沙生针茅没有明显的地理替代分布现象。5种锦鸡儿和7种针茅的分布范围分别又都有一定的重叠。 整理12个物种分布区内的气象台站长期记录,选择计算16个具有重要生物学意义的水热指标值;进而用方差分析、多重比较和因子分析相结合的方法,研究控制这5种锦鸡儿和7种针茅地理分布的主导驱动因子。结果表明:控制小叶锦鸡儿和中间锦鸡儿间地理分布差异的主导因子是水分因子,特别是湿度;水分因子同样是控制中间锦鸡儿和柠条锦鸡儿间地理分布差异的主导因子,特别是生长季及年降水量;控制柠条锦鸡儿和藏锦鸡儿间地理分布差异的主导因子是夏季高温,控制柠条锦鸡儿和狭叶锦鸡儿地理分布差异的是冬季低温。控制贝加尔针茅、大针茅、克氏针茅和戈壁针茅间替代分布的主导气候因子是年降水量和生长季降水量。控制克氏针茅和本氏针茅间替代分布的主导气候因子是温暖指数。 运用耦合BIOCLIM模型的软件包“DIVA-GIS”模拟预测这5种锦鸡儿和7种针茅的现状潜在分布区及未来气候变化的影响。结果表明:现状潜在分布区与实际分布区均有很好的一致性;在CO2浓度加倍的未来气候情景下,这些植物都会向北大幅度迁移,在我国的分布范围均缩小,分布格局发生显著变化。用ROC曲线和Kappa统计值法验证模型表明,BIOCLIM的模拟精度较高。利用BIOCLIM模型绘制了这12个物种的生物气候分室图,并根据生物气候分室确定了物种的最适气候范围。 为了研究锦鸡儿和针茅分布对气候变化的敏感性,本文在现实气象数据的基础上模拟预测了不同降水与温度变化情景下(保持年降水量不变,年均温分别增加1℃、2℃、3℃和4℃;保持年均温不变,年降水量分别增加和减少10%)的物种分布范围,发现随着气温升高和降水量增加,全部锦鸡儿和针茅都会向高纬度地区缓慢迁移,而当降水量减少时,它们将向低纬度地区迁移。不同气候情景下的物种分布范围迁移幅度表明,5种锦鸡儿中狭叶锦鸡儿和中间锦鸡儿的脆弱性相对较大,7种针茅中克氏针茅和贝加尔针茅的脆弱性相对较大。气温的单独变化控制这些物种分布区的未来迁移。 最后,本文探索了锦鸡儿和针茅的气候变化影响的阈值。就核心分布区而言,小叶锦鸡儿、贝加尔针茅、大针茅、沙生针茅的气候变化影响阈值是气温升高4℃,降水减少10%;中间锦鸡儿和狭叶锦鸡儿是气温升高4℃,降水增加10%;柠条锦鸡儿和本氏针茅是气温升高4℃,降水不变;藏锦鸡儿是气温升高2℃,降水增加10%;克氏针茅和短花针茅是气温升高3℃,降水不变;戈壁针茅是气温升高1℃,降水不变。
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Gene number difference among organisms demonstrates that new gene origination is a fundamental biological process in evolution. Exon shuffling has been universally observed in the formation of new genes. Yet to be learned are the ways new exons originate and evolve, and how often new exons appear. To address these questions, we identified 2695 newly evolved exons in the mouse and rat by comparing the expressed sequences of 12,419 orthologous genes between human and mouse, using 743,856 pig ESTs as the outgroup. The new exon origination rate is about 2.71 x 10(-3) per gene per million years. These new exons have markedly accelerated rates both of nonsynonymous substitutions and of insertions/ deletions (indels). A much higher proportion of new exons have Kappa(a)/Kappa(s) ratios > 1 (where K-a is the nonsynonymous substitution rate and K-s is the synonymous substitution rate) than K do the old exons shared by human and mouse, implying a role of positive selection in the rapid evolution. The majority of these new exons have sequences unique in the genome, suggesting that most new exons might originate through "exonization" of intronic sequences. Most of the new exons appear to be alternative exons that are expressed at low levels.
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In this paper, we present the results of purification and characterization of an arginine/lysine amidase from the venom of Ophiophagus hannah (OhS1). It was purified by Sephadex G-75 gel filtration and ion-exchange chromatography on DEAE-Sepharose CL-6B. It is a protein of about 43,000, consisting of a single polypeptide chain. It is a minor component in the venom. The purified enzyme was capable of hydrolysing several tripeptidyl-p-nitroanilide substrates having either arginine or lysine as the C-terminal residue. We studied the kinetic parameters of OhS1 on six these chromogenic substrates. OhS1 did not clot fibrinogen. Electrophoresis of fibrinogen degraded with OhS1 revealed the disappearance of the alpha- and beta-chains and the appearance of lower mel. wt fragments. OhS1 had no hemorrhagic activity. It did not hydrolyse casein, nor did it act on blood coagulation factor X, prothrombin and plasminogen. The activity of OhS1 was completely inhibited by NPGB, PMSF, DFP, benzamidine and soybean trypsin inhibitor, suggesting it is a serine protease. Metal chelator (EDTA) had no effect on it.