以亚硝氮为唯一氮源生长的海洋紫色硫细菌去除无机三态氮

【目的】揭示以亚硝氮为唯一氮源生长的海洋紫色硫细菌去除水体中无机三态氮的特征和规律。【方法】在光照厌氧环境下,以乙酸盐为唯一有机物,在分别以氨氮、亚硝态氮、硝态氮为唯一氮源和三氮共存的模拟水体中,采用Nessler’s试剂分光光度法、N-(1-萘基)-乙二胺分光光度法和紫外分光光度法分别测定水体中氨氮、亚硝态氮和硝态氮的含量,比浊法测定菌体生物量。【结果】随着时间的延长,海洋紫色硫细菌Marichromatium gracile YL28分别在氨氮、亚硝态氮和硝态氮为唯一氮源的水体中对三氮的去除量增加,生物量增大,水体pH升高,并逐渐趋于平衡;YL28对氨氮的最大去除量和最大耐受浓度分别为9.64 mmol/L和36.64 mmol/L,当氨氮浓度低于3.21 mmol/L时,去除率可达97.61%以上;与氨氮相比,以亚硝态氮和硝态氮为唯一氮源,菌体的生长速率、生物量和水体最终pH较低,但对亚硝态氮和硝态氮的去除速率和去除量仍然很高,当亚硝态氮和硝态氮浓度分别达13.50 mmol/L和22.90 mmol/L时,YL28仍能够完全去除。在三氮共存的水体中,YL28也能良好的去除无机三态氮,对亚硝态氮和硝态氮去除能力更强。【结论】在模拟水体中,海洋紫色硫细菌YL28能够分别以氨氮、亚硝态氮和硝态氮为唯一氮源生长,具有良好的耐受和去除无机三态氮的能力,尤其对亚硝态氮具有良好的去除能力。本研究为进一步开发高效脱氮,尤其是去除亚硝态氮的不产氧光合细菌水质调节剂奠定了基础,也为微生物制剂的合理应用提供参考。     

有机碳对海洋着色菌YL28去除无机三态氮的影响

【目的】在无机三态氮(氨氮、亚硝氮和硝氮)共存的模拟海水水体中,阐明有机碳尤其是海藻寡糖对海洋着色菌(Marichromatium gracile)YL28生长及去除无机三态氮的影响规律。【方法】采用次溴酸钠氧化法、N-(1-萘基)-乙二胺分光光度法和紫外分光光度法分别测定水体中氨氮、亚硝氮和硝氮的含量,菌体生物量采用比浊法测定。【结果】在光照厌氧环境中,小分子有机酸盐(乙酸钠、丙酮酸钠、琥珀酸钠和柠檬酸钠)是YL28生长和去除无机三态氮的良好有机碳,亚硝氮、硝氮和氨氮去除率分别达到97.92%、99.98%、73.23%-87.15%。单糖(葡萄糖和果糖)、双糖(麦芽糖和蔗糖)和寡糖(壳寡糖和海藻寡糖)是YL28可利用的有机碳,亚硝氮和硝氮去除率分别达到99%和87%以上,氨氮去除率在44.82%-54.53%之间。多糖(β-环糊精、淀粉、黄原胶、琼脂粉、海藻酸钠和卡拉胶)不是菌体利用和去除无机三态氮的有机碳。酵母提取物可作为菌体生长、去除硝氮和亚硝氮的良好有机碳,但严重抑制氨氮去除。海藻酸钠、β-环糊精和卡拉胶分别与乙酸钠共存时,YL28生长和对无机三态氮去除能力与乙酸钠为唯一有机碳的水平相当。乙酸钠体系中添加海藻寡糖,YL28生长速率、最大生物量以及氨氮的去除速率和最大去除率均升高,添加酵母提取物时,生长速率和最大生物量升高,但氨氮去除速率和最大去除率降低。黑暗厌氧环境下,以乙酸钠和氨氮为唯一有机碳和氮源时,YL28不生长,但在无机三态氮共存时,则能良好生长并去除无机三态氮。【结论】在无机三态氮共存海水体系和厌氧条件下,无论是光照还是黑暗环境,YL28均能良好地生长和去除无机三态氮,小分子有机酸盐(乙酸钠、丙酮酸钠、琥珀酸钠、柠檬酸钠)是其良好的有机碳,相对而言,乙酸钠和丙酮酸钠更好。海藻寡糖与乙酸钠复合可提高菌体生长和脱氮能力。本研究为研制开发高效脱氮微生物制剂及其合理性应用提供了指导。     

亚硝氮对海洋着色菌亚硝氮和氨氮去除以及光合色素合成的影响

【目的】在以亚硝氮为唯一氮源和亚硝氮-氨氮共存体系中,考察和分析海洋着色菌(Marichromatium gracile) YL28菌株对水体亚硝氮的环境适应能力。【方法】采用分光光度法分析亚硝氮、氨氮去除效率以及亚硝氮对菌体生物量和色素含量的影响,采用薄层层析法分析亚硝氮对菌体光合色素组成的影响。【结果】YL28菌株能以亚硝氮为唯一氮源生长,主要积累2种细菌叶绿素(BChl)组分(BChl aTHGG和BChl ap)、1种细菌脱镁叶绿素(Bphe)和玫红品(Rhodopin)、螺菌黄质(Spirilloxanthin)、脱水紫菌红醇(Anhydrorhodovibrin)、番茄红素(Lycopene) 4种类胡萝卜素(Car);YL28生物量和对亚硝氮的去除效率随亚硝氮浓度升高而降低,完全去除亚硝氮的浓度可达200 mg/L以上;当亚硝氮浓度高于25 mg/L,单位质量菌体BChl a和Car总量降低,BChl a和Car合成的末端产物(BChl ap和Spirilloxanthin)以及Bphe相对含量升高,其它4种色素组分相对含量则降低,但Car与BChl a相对含量的比值未见明显变化。当亚硝氮-氨氮共存时,YL28菌株对亚硝氮的耐受能力和去除能力明显提高,完全去除亚硝氮的浓度可达300 mg/L以上;氨氮减缓了亚硝氮对光合色素合成的抑制作用,提高了菌体色素合成总量,各色素组分相对含量的变化与亚硝氮为唯一氮源时的变化规律一致。【结论】YL28菌株能高效去除亚硝氮,亚硝氮对菌株生长和光合色素的合成有抑制作用,但氨氮能明显提高YL28菌株对亚硝氮的适应能力。这为进一步开发高效脱除亚硝氮的APB水质调节剂奠定了基础。     

固氮红细菌(Rhodobacter azotoformans) YLK20去除无机氮的影响因素

【背景】不产氧光合细菌(Anoxygenicphototrophicbacteria,APB)作为一类重要的微生物资源,在水产养殖水体氮污染的修复方面已有广泛研究与应用。养殖水体环境复杂,含多种有机物,尤其是有机氮显著影响菌体除氮功效。【目的】在高浓度无机三态氮(氨氮、硝氮和亚硝氮)共存体系中,阐明小分子有机碳、有机氮和盐度对固氮红细菌(Rhodobacter azotoformans) YLK20去除无机三态氮的影响规律及机制,挖掘针对性强和适应性广的高效除氮菌株。【方法】采用RAST和KEGG方法分析YLK20基因组碳氮代谢途径及耐盐机制;采用次溴酸钠氧化法、紫外和N-(1-萘基)-乙二胺分光光度法分别测定氨氮、硝氮和亚硝氮含量。【结果】基因组显示,YLK20拥有EMP、HMP、TCA、固氮、氨化、氨同化和反硝化碳氮代谢途径,含有soh B、nha C、bet B和gbs A等多种耐盐基因。丙酮酸钠、乙酸钠、柠檬酸钠、乙醇和甘露醇是YLK20生长和去除无机三态氮的良好有机碳,葡萄糖和果糖的存在降低了无机三态氮去除能力,蔗糖体系中硝氮和亚硝氮能被良好去除,但氨氮去除能力较低。在高浓度蛋白胨(3.21 g/L)和尿素(1.43 g/L)体系中,YLK20仍能高效去除无机三态氮。YLK20能在3%盐度内生长良好,低盐度时该菌株能良好去除无机三态氮,高盐度时亚硝氮去除能力受到严重抑制。YLK20对海水和淡水实际养殖水体中的无机三态氮有良好去除效果。【结论】YLK20主要通过氨同化和反硝化途径去除无机三态氮,尤其在高浓度有机氮环境中也能高效去除;该菌株适应盐度范围广,兼可适用于淡水和海水养殖水体;该菌株生长和无机三态氮去除影响因素、规律及除氮机制的阐明,可为APB微生物制剂的合理应用提供指导。     

高脱氮活性海洋着色菌YL28生物膜特性

【背景】细菌生物膜在废水处理领域显示出良好的前景,但目前应用于海水养殖水体处理的菌株主要源自淡水菌株,存在难以适应海水高盐环境的问题。源自红树林的海洋着色菌(Marichromatiumgracile)YL28应用于海水养殖水体处理,不仅具有高效除氮能力,而且趋光贴壁能力很强。【目的】阐明海洋着色菌(Marichromatium gracile) YL28的生物膜形成特性和规律,以期为海水养殖水体生物膜反应系统的开发和应用提供参考。【方法】以生物膜和游离菌体生物量、脱氢酶活性、生物膜多糖含量和蛋白含量、无机三态氮去除活性为测定指标,在光照厌氧环境中研究海洋着色菌YL28菌株的生物膜形成规律、生物活性和脱氮效果。【结果】随着时间延长,4 000 lx光照时游离菌体生物量逐渐升高,但在稳定期前快速降低,而成膜生物量经过延滞期后逐渐升高并趋于稳定,表明培养过程中游离菌体能趋光贴壁生长并形成生物膜。在0-5 000 lx光照范围内培养4 d,低光照强度(500 lx)时成膜率(71.21%)最高,1 000-4 000 lx光照强度下成膜率虽然不是最高(54.64%-68.66%),但适宜菌体成膜,膜生物量干重达到0.60-0.80 mg/cm2。除了5 000 lx光照对成膜菌体脱氢酶活性有不利影响外,成膜菌体和游离菌体脱氢酶活性随光照强度升高而升高,而且没有明显差异。生物膜的形成会导致光反应器内部光照受限,但反应器内部游离菌体的脱氢酶活性并没有降低,由此表明,培养液中的菌体主要在生物膜及其界面生长并游离扩散至培养液中。随光照强度(1 000-5 000 lx)和培养时间(4-10 d)的变化,胞外复合物(Extracellularpolymericsubstances,EPS)中蛋白含量变异较大,多糖含量变化较小;随时间延长,蛋白含量升高,其中3 000 lx时蛋白含量最高;4 000 lx时生物膜菌体与游离菌体脱氮活性相比,单位质量菌体的氨氮和亚硝氮去除活性未受到明显影响,而硝氮去除活性有所降低。【结论】海洋着色菌YL28具有良好的生物膜形成能力,其成膜过程主要是菌体趋光贴壁生长成膜,成膜菌体具有良好的脱氮活性,这为利用生物膜系统消除海水养殖水体氮污染奠定了基础。     

海洋着色菌YL28对含沉积物的海水养殖水体氮污染去除效应的研究

用含有沉积物的氮化物污染实际水体作为测定体系,通过测定水体无机三态氮、总氮、COD、pH以及沉积物总氮、COD和对4种代谢酶活性的动力学过程的监测,评价了一株不产氧光合细菌——海洋着色菌(Marichromatium gracile)YL28对海水实际养殖水体脱氮效应。结果表明:水体本身具有一定的自净作用;与对照组相比,YL28组水体氨氮、亚硝氮、硝氮、沉积物总氮和COD的残留率明显降低(P0.05),在6~10d内水体COD明显升高,水体总氮和pH未见显著差异。处理过程中,在特定的时间范围内,转化酶、蛋白酶、脲酶和脱氢酶活性明显升高(P0.05),与沉积物总氮和COD去除规律相吻合。由此可见,添加YL28对污染海水养殖水体中无机三态氮、沉积物有机氮化物和有机碳化物具有明显的去除作用,本研究为海水养殖微生态水质修复剂的开发和应用奠定基础。     

一株光合细菌的分离鉴定及该菌对氨氮和亚硝态氮的去除作用

【背景】中国是水产养殖大国,氨氮、亚硝态氮是水体中主要的氮源污染物。水体氨氮超标不仅会损伤水生动物的神经系统和肝肾系统,还会导致体表及内脏充血。亚硝态氮过高会阻碍血液运载氧气能力,导致鱼虾缺氧、免疫力下降,从而引发肠炎、烂鳃,甚至窒息死亡。部分光合细菌有去除水体氨氮、亚硝态氮的能力,且对环境友好无二次污染。【目的】从广东养殖水体分离、纯化、筛选出生物活性好的光合细菌(编号SP3)进行种属鉴定,优化培养条件,检测其去除水体氨氮和亚硝态氮的能力,为养殖水体去除氨氮和亚硝态氮提供目标菌株。【方法】用双层平板法从混合菌液中分离得到光合细菌,通过革兰氏染色、碳源利用试验、对无机电子供体的利用试验以及16SrRNA基因序列分析对目标菌株进行种属鉴定;测定菌株SP3在不同pH、不同浓度NaCl条件下的OD600,优化培养条件;通过测定7d内SP3菌株在不同浓度氨氮(氯化铵配制)和亚硝态氮(亚硝酸钠配制)中OD600的变化趋势,确定该菌株对不同氮源的利用情况;用纳氏比色法、分光光度法测定SP3菌株降解水体氨氮和亚硝态氮的能力。Genome walking扩增获得亚硝酸盐还原酶基因(nirS),通过荧光定量PCR研究nirS在氨氮、亚硝态氮去除过程中的表达动态。【结果】筛选出的菌株SP3为革兰氏阴性菌,短杆状;能以醋酸盐、丙酮酸盐、丙酸盐、丁酸盐、乳酸盐、富马酸盐、琥珀酸盐、苹果酸、果糖、葡萄糖作为碳源,不能以乙醇和丙酮作为碳源;能利用硫化钠、硫代硫酸钠、亚硫酸钠作为无机电子供体; 16SrRNA基因序列分析表明其与沙氏外硫红螺菌(Ectothiorhodospira shaposhnikovii)序列相似度为99%;菌株SP3适宜pH为6.0-8.5,适宜盐度为0-3%;菌株SP3以铵盐作为氮源时生长状态明显优于亚硝酸盐;以初始菌液浓度8.6×109CFU/mL、起始氨氮量84.15±0.58 mg/L的条件培养7 d,水体氨氮累计去除量为79.45±0.29 mg/L,氨氮累计去除率达到94.42%;在同样菌浓度和2mg/L亚硝酸钠的条件下培养5d,水体亚硝态氮含量低于检测限0.003mg/L。在菌株SP3去除氨氮、亚硝态氮过程中nirS相对表达量上调。【结论】菌株SP3为沙氏外硫红螺菌(E.shaposhnikovii),能有效去除水中氨氮和亚硝态氮,具有净化水质作用,在水产养殖和污水处理中有广阔应用前景。     

异养硝化-好氧反硝化菌Acinetobacter johnsonii sp. N26的脱氮性能及代谢途径

【背景】水体中含氮物质的大量累积会造成水体富营养化、水生生物死亡等问题,严重威胁水生态环境,制约我国环境保护的持续发展。【目的】为去除生活污水中的含氮污染物,从羊粪堆肥中筛选出了一株具有异养硝化-好氧反硝化功能的细菌——约氏不动杆菌Acinetobacter johnsonii sp.N26,研究其脱氮性能和代谢途径。【方法】测定菌株N26在氨氮和硝态氮中的生长和脱氮曲线,通过单因素试验对其脱氮性能进行优化,通过氮平衡分析和功能基因鉴定研究其脱氮代谢途径。【结果】生长和脱氮曲线表明,菌株N26对初始浓度均为50 mg/L的氨氮和硝态氮的去除速度快、效率高,其中9 h内对氨氮的去除效率为95.5%,最大去除速率为5.330 mg/(L·h);15 h内对硝态氮的去除效率为93.6%,最大去除速率为3.147 mg/(L·h),且最终仅有少量硝酸盐、亚硝酸盐积累。脱氮性能优化结果表明,该菌株的最适氮源为氯化铵,最适碳源为丁二酸钠,最适温度为30℃,最适接种量为15%,最适p H值为8.0-9.0,最适碳氮比为15,最适转速为120 r/min,最适氮负荷≤300 mg/L (氨氮)。氮平衡...     

沼泽红假单胞菌(Rhodopseudomonas palustris)CQV97对无机三态氮共存水体中氮素的去除效率及其影响因素

研究水体环境因素(温度、光照和pH)、小分子有机碳和有机氮化合物对一株具有高效脱氮潜力的沼泽红假单胞菌(Rhodopseudomonas palustris)CQV97在无机三态氮共存体系中脱除无机三态氮的影响规律。结果显示,该菌株在20~40℃,500~5 000lux,pH 6.0~9.0环境条件下,能够脱除高浓度无机三态氮(其中亚硝氮不低于40mg·L-1),表明该菌株具有较强的适应复杂环境的能力;以乙酸钠/乙醇为唯一碳源时,该菌株能有效地去除无机三态氮,而以葡萄糖为唯一碳源时,能有效去除硝氮,但不能去除氨氮,亚硝氮明显积累,表明环境中小分子有机碳源影响菌体对无机三态氮的去除能力;体系中添加高浓度(120mg·L-1)蛋白胨或尿素时,由于有机氮降解的释氨作用,菌体对氨氮的去除能力受到明显抑制,氨氮积累明显,13d时氨氮去除率仅分别为16%(蛋白胨)和6%(尿素),但硝氮和亚硝氮的去除能力并没有受到明显影响。异位处理实际水体结果表明,菌株可使水体中氨氮含量明显降低、硝氮和亚硝氮被完全去除。综上,沼泽红假单胞菌CQV97菌株环境适应能力强,具有高效脱除水体无机三态氮的应用潜力,这为进一步开发高效脱氮微生物制剂及其合理使用奠定了基础。     

4种理化因子对菌株XH1硝化效果的影响

【背景】基于硝化菌群的富集培养技术可高效稳定地去除养殖水体中的有害氮素,而当前在水产养殖领域有关硝化菌群定向培育及硝化功能菌株的研究较少。【目的】研究不同盐度、pH、温度、通气量条件下硝化菌群分离菌株XH1的生长及其对氨氮和亚硝氮的去除效果。【方法】设置不同梯度的盐度、pH、温度、通气量条件,通过计数菌量、测定氨氮及亚硝氮的浓度变化,比较不同条件下菌株XH1的生长及其对氨氮和亚硝氮的影响。【结果】菌株XH1可在盐度5‰-35‰、pH 6.0-9.0、温度15-45°C和通气量0.5-1 V/(V·min)的条件下生长良好,菌量最高可达2.34×109cells/mL;在盐度5‰-35‰、pH 6.0-9.0、温度15-30°C、通气量0.5 V/(V·min)的条件下,对氨氮的去除效果显著(P0.05),在第1-3天对培养液中氨氮的最高去除率可达86%-97%,但培养液中的氨氮浓度先降后升;对亚硝氮的最高去除率达68%。【结论】菌株XH1对盐度、pH、温度等主要环境因子具有良好的适应性,其对水体氨氮的去除效果良好,可作为中低盐度养殖池塘水体氨氮防控菌剂产品研发的备选菌株。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.