四种鳃金龟对不同植物挥发物的触角电位反应

华北大黑鳃金龟Holotrichia oblita Falderman、暗黑鳃金龟Holotrichia parallela Motschulsky、小黄鳃金龟Metabolus flavescens Brenske和福婆鳃金龟Brahmina faldermanni Kraatz有不同的寄主范围.本研究采用溶剂提取法收集寄主植物玉兰Magnolia denudata Desr.、柿树Diospyros kaki L.f.、金银木Lonicera maackii(Rupr.)Maxim、榆树Ulmus pumila L.、芙蓉葵Hibiscus moscheutos L.、丁香Syzygiumr aromaticum(L.)Merrill&Perry和非寄主植物银杏Ginkgo biloba L.等7种植物叶片粗提物,分别测定4种金龟甲雄、雌虫对这些植物粗提物的触角电位反应.结果表明,4种金龟对共同寄主植物金银木和榆树粗提物均有较强的EAG反应;华北大黑鳃金龟雄虫对寄主植物粗提物的EAG反应均弱于雌虫;小黄鳃金龟雌虫对芙蓉葵粗提物的EAG反应强于雄虫;而另外两种金龟雄、雌虫对同种植物粗提物的EAG反应无显著差异.上述结果说明金龟甲的EAG反应大小与田间发生情况一致.     

金龟甲对蓖麻叶挥发物的触角电位和行为反应

为了探明金龟甲偏爱选择有害非寄主植物蓖麻的原因,应用触角电位（EAG）仪和“Y”型嗅觉仪,分别测定了华北大黑鳃金龟、暗黑鳃金龟和铜绿丽金龟雌、雄虫对5种蓖麻叶挥发物的触角电位和选择行为反应。EAG测定结果表明,3种金龟甲雄虫对各处理挥发物的EAG反应值均比参照挥发物顺-3-己烯-1-醇强。华北大黑鳃金龟对邻苯二甲酸二丁酯和肉桂醛的EAG反应值相对较高,暗黑鳃金龟对苯甲醇的EAG反应值相对较高,铜绿丽金龟对邻苯二甲酸二丁酯和苯甲醇的EAG反应值相对较高。选择行为反应测定结果表明,当顺-3-己烯-1-醇与其他挥发物配对测试时,华北大黑鳃金龟对邻苯二甲酸二丁酯和肉桂醛2种蓖麻叶挥发物表现出明显的选择偏好,暗黑鳃金龟对蓖麻叶挥发物苯甲醇表现出明显的选择偏好,铜绿丽金龟则更偏好选择蓖麻的绿叶气味物质顺-3-己烯-1-醇。总之,蓖麻叶的挥发性物质与引诱金龟甲偏爱选择密切相关,且偏爱的挥发物因金龟甲的种类而异。     

华北大黑鳃金龟对桃树挥发物的电生理反应

【目的】华北大黑鳃金龟Holotrichia oblita是一种分布范围广,寄主种类多的农业害虫。本研究鉴定了桃树顶空挥发物中对华北大黑鳃金龟雌虫具有触角电生理活性的化合物,并探索桃树挥发物活性成分的剂量对华北大黑鳃金龟雌、雄虫触角电生理反应的影响。【方法】采用顶空吸附法收集桃树枝叶挥发物,用气相色谱 触角电位联用技术(GC-EAD)和气相色谱 质谱联用技术(GC-MS)鉴定有电生理活性的桃树挥发物成分,最后用触角电位(electroantennograph, EAG)技术测试这些挥发物在0.01,0.1, 1, 10和100 μg剂量下引起的华北大黑鳃金龟雌、雄成虫EAG反应。【结果】桃树挥发物中能引起华北大黑鳃金龟雌成虫触角电生理反应的成分有7种,包括Z-β-罗勒烯、顺-3-己烯乙酸酯、顺-3-己烯醇、壬醛、水杨酸甲酯以及2种未知化合物。所测试的对华北大黑鳃金龟雌成虫触角具有电生理活性的5种桃树挥发物成分在各剂量下都能引起雌、雄成虫明显的EAG反应。雌、雄虫对同种物质的EAG反应随着剂量的增加而有增大的趋势,并且雌虫的EAG反应显著高于雄虫。【结论】桃树挥发物中,罗勒烯、顺-3-己烯乙酸酯、顺-3-己烯醇、壬醛、水杨酸甲酯均能引起华北大黑鳃金龟雌、雄成虫的剂量依赖性的触角电生理反应,而且存在显著的性别差异。可利用这些化合物开展行为反应和田间诱集试验,进一步验证其对华北大黑鳃金龟的引诱活性。     

华北大黑鳃金龟成虫对三种植物的取食选择及趋性行为反应

【目的】本研究旨在明确华北大黑鳃金龟Holotrichia oblita成虫对3种植物的趋向选择性与取食偏好性的关系及其成因。【方法】利用Y型嗅觉仪和叶面积仪等分别测定华北大黑鳃金龟成虫对榆树Ulmus parvifolia、苘麻Abutilon theophrasti和蓖麻Ricinus communis 3种植物的趋向性和取食量,采用GC-MS分析鉴定这3种植物挥发物的成分,观测其对主要成分的嗅觉反应。【结果】华北大黑鳃金龟成虫对蓖麻的趋向性显著强于对苘麻和榆树的趋向性(P0.05),对苘麻和榆树的趋向程度相当;但对榆树叶片的取食量最大(P0.05),对苘麻和蓖麻叶片的取食量相近,雌虫与雄虫的取食量没有显著差异(P0.05)。通过GC-MS分析鉴定发现,3种植物挥发物主要成分有明显差异。嗅觉反应结果显示,在10μg/μL浓度下,顺-3-己烯基异丁酸酯、邻苯二甲酸二丁酯、顺-3-己烯基乙酸酯、芳樟醇、苯酚、水杨酸甲酯和β-石竹烯等化合物对华北大黑鳃金龟的吸引力都显著强于对照(正己烷);但3-蒈烯、反-2-壬烯醛和α-水芹烯对华北大黑鳃金龟的吸引力弱于对照,并呈显著差异(P0.05)。【结论】华北大黑鳃金龟成虫对榆树、苘麻和蓖麻的趋向选择性不等同于取食偏好性。     

竹横锥大象对寄主及虫体挥发物的行为和触角电位反应

为了探明竹横锥大象Cyrtotrachelus buqueti Guerin-Meneville是否存在两性间的引诱作用,以及寄主植物慈竹Neosinocalamus affinis笋对两性行为的影响,采用Y型嗅觉仪测定了竹横锥大象雌、雄成虫对4种虫体及寄主挥发物不同处理（雌、雄成虫,慈竹笋与雌、雄成虫的复合体）的行为反应,测定了触角不同部位对雌雄成虫3种虫体的提取物（整体、鞘翅、后肠）及其与寄主植物挥发物质联合作用的触角电位反应（EAG）。结果表明：雌成虫挥发物对雄虫有较高的引诱活性,对雌虫有驱避作用;雄成虫挥发物对雌虫有一定的引诱作用。雌、雄成虫对虫体与竹笋复合体气味的行为和电生理反应均显著或极显著高于对相应虫体挥发物的反应。经EAG测定,触角各部位对虫体不同部位挥发物的EAG值差异明显,触角端部对虫体与植物挥发性物质的联合作用EAG反应差异极其显著（P＜0.01）。竹横锥大象对沾染雌雄成虫虫体提取物的玻璃棒有一定的兴奋表现,雌虫兴奋率为6.67%,雄虫为26.67%。结果提示,竹横锥大象种内存在信息素,雌虫释放的信息素对同类雄虫有很强的引诱作用,而雄虫所释放的信息素对同类雌虫有引诱作用,添加寄主植物能够增强两性间的引诱效果。     

暗黑鳃金龟对性信息素的触角电生理及行为反应

为了分离鉴定暗黑鳃金龟的性信息素成分并对其功能进行验证,本研究对比摸索了3种触角的处理方法,并进行同一浓度（30 ng·μL^-1）二元混合物、不同配比的触角EAG测试。结果包括: 建立了一种鳃叶状触角的触角电位（EAG）测定方法,即将暗黑鳃金龟Holotrichia parallela触角的各鳃叶用针分离开后进行测定,这种方法测得的触角电位反应值较高。雄虫触角对L-异亮氨酸甲酯和(R)-(-)-芳樟醇为7∶1的二元混合物的反应值最高,和暗黑鳃金龟雌虫性信息素25个腺体提取液的EAG反应相当;对同一配比（7∶1）不同剂量刺激液的EAG测试表明,雄虫对浓度为30 ng·μL^-1的二元混合物刺激液的反应值最高。嗅觉反应结果进一步证实,试虫对L-异亮氨酸甲酯和(R)-(-)-芳樟醇7∶1的选择最高,选择系数达93.3%。研究结果为利用性信息素防治暗黑鳃金龟技术的开发提供了基础。     

暗黑鳃金龟气味结合蛋白HparOBP15a基因的克隆及功能分析

【目的】暗黑鳃金龟Holotrichia parallela通过气味结合蛋白(odorant binding protein,OBP)识别性信息素和植物挥发物准确而迅速地定位配偶、寄主植物。本研究通过克隆暗黑鳃金龟气味结合蛋白15a(Hpar OBP15a)基因,解析该基因的编码蛋白特征、组织表达模式及与寄主植物气味等化合物的结合特性方面的研究,为阐明暗黑鳃金龟基于嗅觉识别的寄主植物选择机理奠定理论基础。【方法】根据暗黑鳃金龟成虫触角转录组测序的结果,利用RT-PCR克隆了Hpar OBP15a基因;Real-time PCR方法分析了该基因在成虫不同部位的表达量差异;荧光竞争结合测定了Hpar OBP15a蛋白和58种候选化合物的结合特征。【结果】暗黑鳃金龟Hpar OBP15a基因全长534 bp,编码147个氨基酸,Gen Bank登录号为AK1834747。Hpar OBP15a在触角中特异表达,且在雌虫触角中表达量显著高于雄虫。在被测的58种化合物中,Hpar OBP15a与46种气味化合物具有较好的亲和性,其中与十二烷、十二醇结合能力最强,其解离常数分别为8.5和11.3μmol/L;同时,对性信息素(L-异亮氨酸甲酯和R-芳樟醇)也有一定的结合能力(解离常数分别为21.0和18.5μmol/L)。【结论】Hpar OBP15a具有广泛的气味结合谱,其中对榆树挥发物十二烷的结合能力最强,因此该蛋白可能在暗黑鳃金龟对榆树的定位过程中具有重要作用。     

华北大黑鳃金龟引诱剂配方筛选及引诱效果评价

【目的】筛选可有效引诱华北大黑鳃金龟Holotrichia oblita成虫的引诱剂配方用于生态防控。【方法】选取5种植物挥发物、2种华北大黑鳃金龟聚集信息素和1种性信息素,根据各化合物单体最佳引诱浓度进行两两混配,得到植物挥发物+聚集信息素、植物挥发物+性信息素和聚集信息素+性信息素共17种二元引诱剂配方。通过触角电位(EAG)反应和趋性行为反应逐步筛选出引诱效果较好的配方。将所筛选配方与其单一组分通过趋性行为反应进行引诱效果比较,并通过室内笼罩诱捕检验配方对华北大黑鳃金龟成虫的引诱效果。【结果】通过EAG反应及趋性行为反应从17种二元引诱剂配方中逐步筛选出了对华北大黑鳃金龟成虫具有较强引诱作用的3种配方,即顺-3-己烯基乙酸酯+顺-9-十八烯乙酸酯,1-己醇+顺-9-十八烯乙酸酯和邻苯二甲酸二丁酯+甘氨酸甲酯,它们引起的华北大黑鳃金龟雌虫EAG反应相对值分别为5 981%, 5 796%和5 722%,引起的雄虫EAG反应相对值分别为3 983%, 4 712%和3 809%。趋性行为反应试验中,引起华北大黑鳃金龟成虫选择反应率最高的为配方邻苯二甲酸二丁酯+甘氨酸甲酯,成虫选择反应率达71.21%,其次为配方1-己醇+顺-9-十八烯乙酸酯和顺-3-己烯基乙酸酯+顺-9-十八烯乙酸酯,成虫选择反应率分别达到70.84%和70.72%,均显著高于其他配方引起的选择反应率。3种二元组分引诱剂配方与其单一组分引诱效果的比较表明,配方顺-3-己烯基乙酸酯+顺-9-十八烯乙酸酯对单一组分顺-3-己烯基乙酸酯具有显著的增效作用,而对顺-9-十八烯乙酸酯增效作用不明显;配方1-己醇+顺-9-十八烯乙酸酯和邻苯二甲酸二丁酯+甘氨酸甲酯均对其单一组分增效作用显著。室内笼罩诱捕结果表明,配方顺-3-己烯基乙酸酯+顺-9-十八烯乙酸酯,1-己醇+顺-9-十八烯乙酸酯和邻苯二甲酸二丁酯+甘氨酸甲酯引诱成虫数均极显著高于石蜡油对照,配方顺-3-己烯基乙酸酯+顺-9-十八烯乙酸酯引起的选择反应率最高,达74.06%;其次为配方1-己醇+顺-9-十八烯乙酸酯和邻苯二甲酸二丁酯+甘氨酸甲酯,引起的选择反应率分别达到65.89%和64.25%;但3种配方引起的华北大黑鳃金龟成虫的选择反应率之间无显著性差异。【结论】顺-3-己烯基乙酸酯+顺-9-十八烯乙酸酯,1-己醇+顺-9-十八烯乙酸酯和邻苯二甲酸二丁酯+甘氨酸甲酯这3种二元引诱剂配方对华北大黑鳃金龟成虫均具有较强的引诱作用,均可作为用于生态治理华北大黑鳃金龟的有效引诱剂。     

白星花金龟聚集信息素的分离鉴定及引诱效果

[目的]明确白星花金龟Protaetia brevitarsis Lewis聚集信息素的有效成分,为白星花金龟的绿色防控提供支持.[方法]通过动态顶空法收集白星花金龟雌虫、雄虫和雌雄虫混合挥发物,用气相色谱-质谱(Gas chromatography-mass spectrometry,GC-MS)联用技术对收集的挥发物进行分析鉴定,并利用触角电位(Electroantennogram,EAG)仪和Y型嗅觉仪测定白星花金龟对8种化合物的触角电位和行为选择反应.[结果]白星花金龟雌虫、雄虫和雌雄虫混合挥发物中皆鉴定出8类15种化合物,包括6种烷烃类、2种酚类、2种芳香烃类、1种烯类、1种吡啶类、1种胺类、1种醇类和1种酮类.EAG试验结果表明,白星花金龟对8种化合物都有EAG反应,且不同挥发物间、同种挥发物不同浓度间的EAG反应存在差异.白星花金龟对蒎烷胺、3,4-二甲基-6-乙基苯酚、2,3-二甲基苯乙酮和2,6,10,14-四甲基十七烷4种挥发物的EAG反应相对值在1μg/μtL时达到最大,对萘、苯并噻唑、百里酚和正十四烷4种挥发物的EAG反应相对值在10 μtg/μtL时达到最大.行为选择反应表明,白星花金龟雌成虫、雄成虫对2,3-二甲基苯乙酮有明显趋向性,选择反应率大于70％,对2,6,10,14-四甲基十七烷、萘、苯并噻唑、百里酚和正十四烷5种物质均表现出显著或极显著的驱避效果,对3,4-二甲基-6-乙基苯酚和蒎烷胺表现出了性别上的差异,雄虫对3,4-二甲基-6-乙基苯酚表现出显著的趋向性,对蒎烷胺表现出显著的驱避性,而雌虫对这2种挥发物却无明显反应.[结论]2,3-二甲基苯乙酮对白星花金龟雌成虫、雄成虫有明显的引诱效果,2,6,10,14-四甲基十七烷、萘、苯并噻唑、正十四烷和百里酚有较强的驱避作用,推测2,3-二甲基苯乙酮是白星花金龟聚集信息素的有效组分.     

牧草盲蝽对七种寄主植物不同挥发物的EAG和行为反应

【目的】研究牧草盲蝽Lygus pmtensis对寄主植物挥发物的EAG及趋向行为反应,为研发牧草盲蝽引诱剂或趋避剂策略提供科学依据。【方法】利用固相微萃取方法收集7种寄主植物包括木地肤Kochia prostrata (L.) Schrad.、灰绿藜Chenopodium glaucum、花椰菜Brassica oleracea L.var.botrytis L.、小白菜Brassica campestris L.ssp.chinensis Makino、田旋花Convolvulus arvensis L.、小藜Chenopodium serotinum L.、番茄Lycopersicon esculentum Mill.的挥发物,通过室内气相-质谱联用系统(GC-MS)进行挥发物的定性及定量分析,最终测定了9种候选特征挥发物的触角电位(EAG)反应和嗅觉趋向行为反应。【结果】7种寄主植物共鉴定到的化合物有31种,且种类和含量有较大差异,差异显著。9种候选特征挥发物均能引起牧草盲蝽雌、雄成虫的EAG反应。其中牧草盲蝽雌、雄成虫对壬醛的EAG反应绝对值最大,雌、雄虫对不同植物挥发物质EAG反应相对值均有显著差异,且对同种化合物雌虫的反应相对值均大于雄虫的反应,乙酸顺式-3-己烯酯和壬醛引发的EAG反应值差异显著;趋向行为测定中,牧草盲蝽雌虫对壬醛、金合欢烯、α-蒎烯、1-石竹烯、乙酸顺式-3-己烯酯、2-甲基-1-醇的大部分浓度的行为反应差异显著,雄虫仅对α-蒎烯、壬醛的部分浓度的行为反应差异显著。【结论】7种寄主植物的挥发物对牧草盲蝽雌、雄成虫具有不同EAG和趋向行为反应影响,候选气味化合物可以作为设计牧草盲蝽引诱剂组分进行田间害虫防控实践。     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Synthesis and turnover of cerebrosides and phosphatidylserine of myelin and microsomal fractions of adult and developing rat brain.

The synthesis and turnover of cerebrosides and phospholipids was followed in microsomal and myelin fractions of developing and adult rat brains after an intracerebral injection of [U-14C]serine. The kinetics of incorporation of radioactivity into microsomal and myelin cerebrosides indicate the possibility of a precursor-product relationship between cerebrosides of these membranes. The specific radioactivity of myelin cerebrosides was corrected for the deposition of newly formed cerebrosides in myelin. Multiphasic curves were obtained for the decline in specific radioactivity of myelin and microsomal cerebrosides, suggesting different cerebroside pools in these membranes. The half-life of the fast turning-over pool of cerebrosides of myelin was 7 and 22 days for the developing and adult rat brain respectively. The half-life of the slowly turning-over pool of myelin cerebrosides was about 145 days for both groups of animals. The half-life of the rapidly turning-over microsomal cerebrosides was calculated to be 20 and 40 h for the developing and adult animals respectively. The half-life of the intermediate and slowly turning-over microsomal cerebrosides was 11 and 60 days respectively, for both groups of animals. The amount of incorporation of radioactivity into microsomal cerebrosides from L-serine was greatly decreased in the adult animals, and greater amounts of the precursor were directed towards the synthesis of phosphatidylserine. In the developing animals, considerable amounts of cerebrosides were synthesized from L-serine, besides phosphatidylserine. The time-course of incorporation indicated that a precursor-product relationship exists between microsomal and myelin phosphatidylserine. The half-life of microsomal phosphatidylserine was calculated to be about 8 h for the fast turning-over pool in both groups of animals.     

Identification and composition of the tonsillar and anal enterococcal and streptococcal flora of dogs and cats

Enterococcus faecalis was the most frequently isolated enterococcal species from anal swabs and tonsils of dogs and cats, although in the anal samples from dogs Ent. hirae was found almost as often as Ent. faecalis. Most Ent.faecium strains from dog tonsils differed from those associated with humans and other animals in that they fermented sorbitol. Typical Ent. avium as well as atypical Ent. avium -like strains were seen in dogs, while the related species Ent. raffinosus was associated with cat tonsils. Enterococcus cecorum also occurred mainly in cats. Certain atypical strains, presumptively identified as Ent. cecorum , shared characteristics with Ent. columbae.
The most frequent streptococcal species in tonsils of cats and dogs were Streptococcus suis and Strep. canis. Streptococcus canis and Strep. bovis predominated in anal swabs. The canine Strep. suis differed from the common porcine strains in fermenting mannitol.
Forty-seven of the 288 isolates examined could not be identified or related to known species. The characteristics of two groups of these bacteria, provisionally called 'Ton 31 group' and 'O7 group' are described.     

Molecular characterisation of species and genotypes of Cryptosporidium and Giardia and assessment of zoonotic transmission

The molecular characterisation of species and genotypes of Cryptosporidium and Giardia is essential for accurately identifying organisms and assessing zoonotic transmission. Results of recent molecular epidemiological studies strongly suggest that zoonotic transmission plays an important role in cryptosporidiosis epidemiology. In such cases the most prevalent zoonotic species is Cryptosporidium parvum. Genotyping and subtyping data suggest that zoonotic transmission is not as prevalent in the epidemiology of giardiasis. Molecular characterisation of Cryptosporidium and Giardia is a relatively recent application that is evolving as new genes are found that increase the accuracy of identification while discovering a greater diversity of species and yet unnamed taxa within these two important genera. As molecular data accumulate, our understanding of the role of zoonotic transmission in epidemiology and clinical manifestations is becoming clearer.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000